pubs.acs.

org/materialsau Review

Combinational Gene Therapy toward Cancer with Nanoplatform:

Strategies and Principles
Jinhui Lin, Xinlian Wang, Dongqi Ni, Yandong Chen, Chunying Chen, and Ying Liu*

Cite This: https://doi.org/10.1021/acsmaterialsau.3c00035 Read Online

ACCESS Metrics & More Article Recommendations

See https://pubs.acs.org/sharingguidelines for options on how to legitimately share published articles.

ABSTRACT: Cancer remains a significant threat to human health. While

numerous therapies have been developed to combat the disease,
traditional treatments such as chemotherapy and radiotherapy are
suboptimal and associated with significant side effects. Gene therapy is
Downloaded via 2.147.12.71 on October 26, 2023 at 09:16:40 (UTC).

an emerging therapeutic approach that offers improved targeting and

reduced side effects compared with traditional treatments. Using siRNA
and other nucleic acid-based drugs in cancer treatment has generated
significant interest among researchers. Nanocarriers, such as liposomes,
can effectively deliver these agents to tumor sites. However, gene therapy
alone is often insufficient to eradicate tumors, and there is a risk of
recurrence. Therefore, combining gene therapy with other therapies using
nanocarriers, such as phototherapy and magnetic hyperthermia therapy,
can lead to synergistic therapeutic effects through different mechanisms.
In this review, we summarize various ways in which gene therapy can be combined with other therapies and highlight the role of
nanoplatforms in mediating these combined therapies, which would inspire novel design ideas toward combination therapies.
Additionally, bottlenecks and barriers to gene therapy should be addressed in the near future to achieve better clinical efficacy.
KEYWORDS: combinational therapy, gene therapy, tumor treatment, nanoplatform, chemotherapy, phototherapy,
magnetic hyperthermia therapy, immune therapy

1. INTRODUCTION The nanodrug delivery system provides an excellent

Cancer, a major public health problem worldwide, has tended
to be the leading cause of death in the 21st century. According
to the American Cancer Society estimates, nearly 600,000
people in the United States will die because of cancer in 2022.1
The number of Americans with cancer history is estimated to
be more than 22.1 million in 2030 based on the growth and
aging of the population alone.2 Chemotherapy, radiotherapy,
and surgery are the main treatments for tumors, but their
effects are not ideal for high mortality and side effects clinically.
Meanwhile, tumor recurrence and metastasis are still a great
challenge clinically, while there are no suitable therapeutics.
After almost 30 years of development, gene therapy is
becoming one of the most widely used therapeutic strategies
except traditional therapies against cancers and can import
healthy exogenous or therapeutic genes into host cells to kill
cancer cells or protect normal cells.3 Several types of nucleic
acid drugs were explored to heal cancer, such as Bcl-2 siRNA
and suicide gene. Nucleic acid drugs can be divided into DNA
and RNA, interfering with or regulating targeted gene
expression in specific cells. Most types of DNA drugs target
the cell nucleus, while RNA drugs target the cytoplasm.
However, gene drugs can be easily degraded in vessels and into
other organs; therefore, gene drugs are limited clinically.
© XXXX The Authors. Published by
American Chemical Society
A ACS Mater. Au XXXX, XXX, XXX−XXX
platform for the delivery of gene drugs with its protective
properties and targeted effects. Through rational design,
nanomaterials can encapsulate nucleic acids effectively and
protect gene drugs from degradation. Moreover, some types of
nanomaterials could help gene drugs escape from lysosome
and codelivery other drugs with nucleic acid. Many nano-
vectors are developed to deliver gene drugs, including
liposomes, cationic polymers, etc. Nowadays, novel therapies
are urgently needed for cancer treatments. Combining gene
therapy with the currently available therapies such as
chemotherapy or radiation therapy provides surprising hope
for better therapeutic effects in patients with advanced-stage
cancers. Nanovectors, which can load more drugs onto
targeted organs and deliver them into the cytoplasm, perform
excellently in combined gene therapy. Based on the therapeutic
effect of gene delivery, we summarize the purpose and effects
Received: April 29, 2023
Revised: July 19, 2023
Accepted: July 20, 2023
https://doi.org/10.1021/acsmaterialsau.3c00035
ACS Materials Au pubs.acs.org/materialsau Review

Figure 1. Synergistic therapeutic effect of chemotherapy and gene therapy. (A) Scheme of chemotherapy and gene therapy. (B) Nanocomplex
could inhibit the expression of PIKM2, induce mitochondrial exhaustion and collapse, and enhance the cytoplasmic levels of cytochrome C.
Reprinted with permission from ref 22. Copyright 2021, Elsevier B.V. (C) PAP could eliminate the tumor and suppress the expression of HIF-α.
Reprinted with permission from ref 26. Copyright 2018, Elsevier B.V. (D) MDSCs were stained with CD11b and Gr-1 antibody and determined by
flow cytometry. Reprinted with permission from ref 32. Copyright 2019, Elsevier B.V. (E) CCP/PCBP2 siRNA nanocomplex resulted in significant
tumor inhibition via collagen expression. Reprinted with permission under a Creative Commons Attribution 4.0 International License from ref 36.
Copyright 2021, Ivyspring International Publisher.

of gene therapy in combined therapy with a nanoplatform and
the mechanism of combined therapy.
2. GENE THERAPY FOR TUMORS
2.1. Advantages of Gene Therapy
The main methods of gene therapy against cancer include
using plasmid DNA containing therapeutic nucleotides to
modulate aberrant gene expression and RNA interference like
siRNA, shRNA, miRNA, and antisense oligonucleotide (ASO)
B https://doi.org/10.1021/acsmaterialsau.3c00035
to reduce cancer-related protein expression.4 Compared to
conventional treatments, gene therapy offers a variety of
advantages for cancer treatment, including high potency and
specificity, low off-target toxicity, and delivery of multiple
genes that concurrently target cancer tumorigenesis, recur-
rence, and drug resistance.5,6 As a powerful tool, genome
editing technologies have been used widely in gene therapy for
their properties of gene addition, gene ablation, and gene
“correction” in vivo or in vitro, different from viral vectors that
ACS Mater. Au XXXX, XXX, XXX−XXX
ACS Materials Au pubs.acs.org/materialsau
can only mediate gene addition.7 For example, adoptive cell
therapy uses gene delivery to ex vivo modify the patient’s T
cells to generate specific antitumor reactivity. It can improve
the antitumor immune response in the body through artificial
methods, achieving a sustained and strong antitumor effect.
Until now, chimeric antigen receptor T cell (CAR-T) therapy,
one of the adoptive cell therapies, has been developed to treat
hematologic malignancies and multiple solid tumor types.8
Suicide gene therapy is based on introducing a suicide gene
into specific tissue, such as a tumor, and triggering cell death
by expressing specific enzymes, toxins, or pro-apoptotic
proteins in tumor cells.9,10 It can enhance the sensitivity of
tumor cells to chemotherapy and be controlled by a tumor-
specific promoter to select expression in tumor cells.11
2.2. Current Challenges of Gene Therapy
There are many barriers inside and outside the cell for nucleic
acid delivery. Naked nucleic acid can be easily degraded in
blood for the presence of nucleases and has a very short half-
time in physiological media. Once nucleic acid gets into a cell,
it must escape from the lysosome or will be degraded in the
lysosome. The tumor microenvironment (TME) can exhibit
abnormal conditions, including hypoxia, low pH, and high
reactive oxygen species (ROS) levels. Following single therapy,
these conditions can promote cancer resistance, tumor
aggressiveness, metastasis, and recurrence.
Moreover, some types of cells in tumor-like cancer stem cells
(CSC), hypoxic cells, and S-phase cells have much more robust
resistance to chemotherapy or radiation therapy, which tend to
induce cancer recurrence or metastasis.12 For solid tumors, it is
difficult for nucleic acid drugs to get into tumors and transfect
target cells because of the complex compounds in solid tumors.
The unfavorable physicochemical properties of nucleotides,
such as negative charges, large molecular weight, and size,
seriously hinder their entry into host cells. Meanwhile, clinical
gene therapeutics are limited by the low stability of nucleotides
induced by endonuclease in serum.12,13 Therapeutic nucleo-
tides must overcome various biological barriers, from their
entry into the bloodstream to their final destination in cancer
cells to reach the cancerous site of action. For example, while
injecting plasmid complex by intravenous injection, the gene
complex should overcome the following biobarriers: (1)
degradation by serum enzymes, (2) being removed by
phagocytes and organs like kidney and liver, (3) crossing the
vascular endothelial cells, (4) tumor microenvironment, (5)
crossing the cellular membrane, (6) escape from endosome,
and (7) crossing the karyotheca.13,14 Nanocarriers as
promising tools could help nucleic acid drugs overcome the
biobarriers mentioned above. Several types of nanocarriers
could be stable in normal tissues and blood and degraded in
tumors by the ligands or properties of nanomaterials. Other
nanocarriers may help drugs cross vascular cells and escape
from cell endosomes. There are some types of nanocarriers
currently used for gene therapy, including liposomes,
polymeric nanoparticles, dendrimers, and viral vectors. Lip-
osomes are spherical vesicles made of phospholipids that can
encapsulate and deliver DNA or RNA. Polymeric nanoparticles
are made of polymers that can bind to nucleic acids.
Dendrimers are highly branched, tree-like polymers that can
encapsulate and deliver DNA or RNA. Viral vectors are
genetically engineered viruses that can deliver therapeutic
genes to cells. Each type of nanocarrier has its advantages and
limitations, and researchers are continually working to improve
C https://doi.org/10.1021/acsmaterialsau.3c00035
Review
their efficiency and safety, as well as exploring the development
of novel nanocarriers.
3. COMBINED GENE THERAPY WITH OTHER CANCER
THERAPIES FOR BETTER THERAPEUTIC EFFECT
Single therapies can hardly realize the desired therapeutic
outcomes in clinical trials because of the unfavorable drug
resistance and heterogeneity of tumors. Since then, synergistic
combination therapies combining two or more therapeutic
modalities have been receiving more and more attention. In
cancer therapy, gene therapy is multifunctional, including the
induction of cancer cell apoptosis, inhibition of tumorigenesis
or metastasis, downregulation of heat shock proteins, and
induction of immune response by expressing cytokines.
Nowadays, it is becoming a strong complement to other
cancer therapy. With the rapid development of nanometer
material science, various nanomaterials with the ability to kill
tumor cells and deliver gene drugs have been widely
developed. These nanomaterials can use magnetic, luminous,
or sound energy to kill cancer cells and deliver therapeutic
nucleic acids like siRNA, ASOs, and so on to gain a much
better therapeutic effect.
3.1. Combined Gene Therapy with Chemotherapy
Chemotherapy is the most common method of cancer
treatment. The cell’s metabolism can be disturbed or interfered
with by delivering chemical drugs into the cell, thus killing
cells. However, chemotherapy is greatly limited by multidrug
resistance, high drug dose concentration, low drug perme-
ability, and adverse effects.15,16 To improve the antitumor-
igenicity, gene therapy is considered to supplement synergistic
chemotherapy. Gene therapy works in conjunction with
chemotherapy to increase the death of tumor cells or overcome
the above-mentioned mechanisms and break the barriers to
chemotherapy in the below ways (Figure 1A).
3.1.1. Synergistic Therapeutic Effect of Chemo-
therapy and Gene Therapy. By design of an intelligent
drug delivery platform, chemotherapy and gene therapy can
work together to heal cancer. Several gene medicines have
been found and used in trials with chemotherapy treatment.
The tumor necrosis factor-related apoptosis-inducing ligand
(TRAIL), a member of the tumor necrosis factor family, can
selectively induce cancer cell apoptosis by binding to the death
receptor 4 (DR4), and DR5 is overexpressed in cancer cells.17
Besides, the TRAIL-based gene therapy produces the by-
stander effect to inhibit tumor growth in vivo.18 However, the
following TRAIL-resistance of cancer cells undermines the
TRAIL-based gene therapy.19 Human antigen R (HuR) is a
potential cancer therapy target associated with tumorigenesis
and metastasis. Amreddy et al. developed a folate receptor-
α(FRA)-targeted polyamidoamine dendrimer-based nanopar-
ticle system (Den-PEI-CDDP-siRNA-FA) for chemo/gene
combined therapy.20 They used Den-PEI-CDDP-siRNA-FA to
codeliver anti-HuR siRNA and cis-diamine platinum to treat
FRA-overexpressing lung cancer cells. They gained a
significantly more significant therapeutic effect than individual
therapeutics. The enhanced mechanism of combined gene
therapy with chemotherapy can be divided into the following
content.
3.1.2. Fighting Multidrug Resistance in Treating
Tumor. The use of chemotherapy, one of the most widely
used and effective treatments for cancers, is seriously limited
by multidrug resistance (MDR), which is developed by the
ACS Mater. Au XXXX, XXX, XXX−XXX
ACS Materials Au pubs.acs.org/materialsau
continuous use of chemotherapy drugs. The cancer MDR
requires an increasing dose of chemotherapy drugs, which
could induce inevitable toxicity to patients. MDR is closely
related to the overexpression of transporters like P-
glycoprotein (P-gp) on the cancer cell membrane. Many
drugs, such as doxorubicin (DOX) and paclitaxel (PTX), must
enter the cells to play a therapeutic role. At the same time, P-
gp, one of the ATP-binding cassette (ABC) transporters, can
actively expel them out of the cells, reducing the efficacy.21
One of the strategies against cancer MDR is blocking out the
energy supply for ATP-binding cassette (ABC) transporters,
achieving a favorable therapeutic effect. A polyelectrolyte
nanocomplex surface decorated with hyaluronic acid was
developed to deliver DOX and PKM2 siRNA.22 The
hyaluronic acid of the nanocomplex could target CD44,
which is overexpressed in tumor cells, while the nanocomplex
could efficiently escape from lysosomal entrapment and release
siRNA into cells. By downregulation of PKM2, the energy
supply for ABC transporters in cancer cells has been
suppressed, thus defeating drug resistance. Results showed
that combined therapy could enhance and decrease the
expression of PKM2, induce mitochondrial exhaustion and
collapse, and release cytochrome C into the cytoplasm (Figure
1B). Another mechanism of MDR is overexpressing growth
factors in cancer cells.23 RNase-resistant RNA nanoparticles
are modified with an epidermal growth factor receptor (EGFR)
to block the growth factor signing pathway. Targeting aptamer
loading XBP1 siRNA, which could promote sensitization to
chemotherapy and impede angiogenesis in vivo, was developed
to kill cancer cells and inhibit tumor growth.24 Hypoxia is a
hallmark feature of the tumor microenvironment, which could
inhibit the efficiency of chemotherapy.25 The aim is to reduce
hypoxia’s influence, a novel multifunctional hypoxia-induced
size-shrinkable nanoparticle designed to codelivery DOX and
si-HIF1α.26 The nanoparticle could respond to hypoxia and
degrade the outer PEG to expose positively charged PAMAM,
thus penetrating DOX and si-HIF1α into the tumor core. The
expression of HIF1α induced by hypoxia is downregulated by
si-HIF1α, which ultimately enhances the effectiveness of DOX
in treating cancer (Figure 1C).
3.1.3. Restoring Homeostatic Stromal Function to
Promote Therapeutic Efficiency and Drug Delivery.
Besides the MDR of cancer cells, tumor stroma also plays an
important role in drug resistance.27 To delay the MDR
progression, optimized combined treatments are badly
needed.28 The tumor-associated macrophages (TAMs) con-
tribute to drug resistance and relapse via different pathways
and are major components of the tumor microenvironment.29
The hyaluronic acid-based nanoparticles were designed to
encapsulate miR-125b (HA-PEI-miR-125b), which can specif-
ically target TAMs and repolarize macrophages to an immune-
activating phenotype. Results show that the total number of
CD11b+, F4/80+, and CD206+ macrophages was reduced
while increasing the number of CD80+ macrophages. However,
there is no obvious difference of CD45+ immune cells between
the untreated control and HA-PEI-miR-125b treated group,
and whether the nanoparticle could increase the number of
PD-1+ immune cells by increased infiltrating T cells needs
further evaluation. Later results show that HA-PEI-miR-125b
could enhance the efficacy of paclitaxel with low toxicity.30
Myeloid-derived suppressor cells (MDSC) are largely
immature myeloid cells that inhibit immune effector cell
function in cancer and tumor progression, representing a
D https://doi.org/10.1021/acsmaterialsau.3c00035
Review
potential therapeutic target for cancer therapy.31 A multifunc-
tional delivery system based on an amphiphilic polymer with
morpholine attached to the pendant side chains (POEG-st-
Pmor) for codelivery of IL-36γ expression plasmid and
doxorubicin (DOX) has been developed to treat cancer. The
nanomicelles are stabilized by charge−charge interactions
between the cationic polymer and plasmid DNA, which are
further addressed in vivo. The Dox+IL-36γ/POEG-st-Pmor
decreases the MDSC in the lung metastasis of breast cancer
and hence enhances the type I immune response, which raises
the efficiency of DOX (Figure 1D).32
One of the main difficulties in treating tumors such as
pancreatic cancer is the nearly impenetrable desmoplastic
stroma. It prevents drugs into tumors and supports and
promotes pancreatic cancer at the same time.33,34 One strategy
to achieving more drugs into stroma-rich tumors is restoring
homeostatic stromal function. A gene-/chemo-combined
strategy was developed.35 The PEGylated PEI-coated gold
NPs were developed to codeliver anti-HSP47 siRNA and
ATRA to transform activated pancreatic stellate cells into
quiescent pancreatic stellate cells, which could promote
gemcitabine delivery into a pancreatic tumor. A peptide-
based core-stabilized PCBP2 siRNA nanocomplex was
developed to break type I collagen in pancreatic ductal
adenocarcinoma (PDAC) tumor stroma (Figure 1E).36 After
siRNA reverses the accumulation of type I collagen, the
antitumor efficacy of gemcitabine was significantly improved.
Cancer-associated fibroblasts could secret a dense desmoplastic
matrix or release regulatory molecules, thus enhancing
chemotherapy resistance.37 Blocking collagen cross-linking
and fibronectin fibril assembly via lysyl oxidase (LOX)-
siRNA could enhance doxorubicin sensitization against the
triple-negative breast cancer (TNBC) cell, indicating that
combining DOX with LOX siRNA is a promising therapeutic
method.38
While chemotherapy is a commonly used method for
treating cancer, it has limitations such as multidrug resistance,
high drug doses, low drug permeability, and adverse effects.
However, gene therapy can be used with chemotherapy to
increase the death of tumor cells or overcome these limitations.
By the design of intelligent nanomaterials, the synergistic
therapeutic effect of chemotherapy and gene therapy can be
achieved in two ways. One way is to combat multidrug
resistance in tumors by disrupting gene expression within the
tumor cells. Another way is by restoring the homeostatic
stromal function, which can also promote therapeutic
efficiency and drug delivery. Finally, a combined gene/chemo
strategy can effectively treat tumors.
3.2. Combined Gene Therapy with Phototherapy
Phototherapy can be divided into two main categories:
photothermal therapy (PTT) and photodynamic therapy
(PDT). PTT is a widely developed localized cancer therapy
that utilizes photoinduced heat via near-infrared irradiation to
cause cancer cell death and avoid damage to other regions
which are not irradiated.39−41 Compared to traditional
therapies, the main advantages of PTT are minimal
invasiveness, deep tissue penetration, and simple opera-
tion.40,42 Another promising cancer therapy based on photo-
sensitizer (PS) is PDT, which uses visible light to irradiate PS
to generate ROS, thereby killing cells.40 PDT is a good choice
for clinical cancer treatment, particularly for superficial tumors
like esophageal, bladder cancer, and melanoma, because of its
ACS Mater. Au XXXX, XXX, XXX−XXX
ACS Materials Au pubs.acs.org/materialsau Review

Figure 2. Synergistic therapeutic effect of phototherapy and gene therapy. (A) Scheme of phototherapy and gene therapy. (B) R-nano ICG/S-
siRNA could destroy tumor blood vessels and increase tumor temperature. Mice injected with PBS (1), ICG (2), nanoICG (3), R-nanoICG (4),
and R-nanoICG/S-siRNA (5) and irradiated with an 808 nm laser at a power density of 1.0 W cm−2. Reprinted with permission from ref 49.
Copyright 2018, Wiley-VCH. (C) Representative noninvasive imaging of lung-adenocarcinoma NCI-H889 tumor-bearing nude mice with different
treatments at days 7 and 36 and immunohistochemical analysis of VEGF in lung tissue. Reprinted with permission under a Creative Commons
Attribution 4.0 International License from ref 60. Copyright 2021, Springer Nature. (D, E) DOX-siVEGF-NPs/Ce6-MBs generate more ROS with
laser and inhibit tumor growth. Reprinted with permission from ref 80. Copyright 2020, Elsevier B.V. (F, G) PEG and PEI dual-functionalized BP
nanosheets could target tumor sites and induce cell death. Reprinted with permission from ref 86. Copyright 2018, American Chemical Society.

E https://doi.org/10.1021/acsmaterialsau.3c00035
ACS Mater. Au XXXX, XXX, XXX−XXX
ACS Materials Au pubs.acs.org/materialsau
eximious features such as low invasiveness, mild killing,
spatiotemporal selectivity, and lower side effects.43,44 One of
the most significant advantages of phototherapy is its ability to
control drug release achieved by visible or infrared light.
However, for those metastatic or disseminated cancers, a single
PTT or PDT appears powerless.45
Currently, gene therapy is an effective complement to PTT/
PDT because of its superiority in therapeutic enhancement and
decreasing side effects.46−49 Therapeutic nucleic acids, such as
siRNA, have been widely used to combine PTT/PDT to gain a
synergistic therapeutic effect. The synergistic effect of gene
therapy and PTT is mainly achieved in two ways. Either gene
therapy and PTT kill tumor cells together, in which gene
therapy can kill the surrounding or PTT-resistant tumor cells,
or gene therapy can improve the sensitivity of tumor cells to
PTT, such as inhibiting the expression of heat shock proteins
from improving the efficacy of PTT. Several inorganic
materials have been applied in gene/photothermal combined
therapy, such as gold nanoparticles (GNs), black phosphorus
nanosheets (BP NS), carbon-based materials, and
others.39,50,51 Some inorganic nanomaterials cannot be loaded
with therapeutic nucleic acids, so modification of these
materials is needed, such as cationic polymer modification
on the material’s surface to load the nucleic acid by
electrostatic interaction between the cationic polymer and
the nucleic acid.41
3.2.1. Synergistic Therapeutic Effect of Phototherapy
and Gene Therapy. The most widely used strategy for gene
therapy combined with PTT/PDT is the synergistic method
for utilizing antitumor genes with PS together (Figure 2A).
Survivin, a member of the inhibitor of apoptosis (IAP) families,
is overexpressed in most human tumors and is considered an
excellent target for cancer gene therapy.52,53 Metal−organic
nanostructures (MONs) have significant advantages in
combination therapies for traditional photosensitizers like
indocyanine green (ICG) with therapeutic nucleotide.49,54 The
Zn(II)-dipicolylamine (Zn-DPA) molecule is a potential
siRNA delivery carrier, because of its high affinity for
phosphate-containing molecules. Gold nanorods (GNRs)
conjugated with Zn-DPA were designed to deliver PLK1-
siRNA and PTT simultaneously. They found that the
combination of PLK silencing-induced apoptosis and local
PTT had apparent synergy for treating PC-3 tumor.46 A
coordination-driven self-assembly MONs have been developed
for gene/photothermal combined therapy.49 The Zn(II)-
dipicolylamine (Zn-DPA) molecule interacted with ICG and
polyvinylpyrrolidone (PVP) polymers to form nanoICG,
which was modified with RGD, one of the peptide motifs
targeting angiogenesis. Due to the high siRNA transfection,
long-lasting tumor accumulation, and good photothermal
properties, the R-nanoICG binding with survivin-siRNA
presented apparent synergistic efficacy of PTT and gene
therapy. The angiogenesis assay showed that the PTT/gene
therapy group had the lowest level of CD31, which indicated
that combination therapy had the most efficiency for
destroying tumor blood vessels (Figure 2B). In the 4T1
xenograft mouse model, the tumors were entirely eliminated
after combination therapy treatment, while the tumors could
not be inhibited by a single therapy.
BP NS, a kind of attractive two-dimensional (2D) material
with photodegradable character, high surface area, and negative
charge, has high photothermal conversion efficiency and
significant extinction coefficient, considered a good nanoma-
F https://doi.org/10.1021/acsmaterialsau.3c00035
Review
terial for PTT.39,55,56 Moreover, BP NS has been efficient PTT
therapy agents for anticancer treatment because of its ability to
generate singlet oxygen (1O2) with a high quantum yield of
∼0.91 under the whole range of visible light.56 In 2018, Wang
et al. first utilized BP NS for gene-/PTT-combined therapy and
reported it was a promising tool for combined therapy.57 The
negatively charged BP NS was modified with PEI, which
enabled it to be loaded with survivin-siRNA. The results of the
Western blot showed that the expression level of survival in
MCF-7 cells (breast cancer cells) was significantly decreased
by being treated with BP-PEI-siRNA for 24 h, which could
inhibit cell growth (44% inhibition rate). With simple design,
the BP nanosheets could degrade into phosphate ions in vivo,
which indicates that the BP-PEI-siRNA complex could be a
promising tool for clinical applications.
Furthermore, the inhibition rate of MCF-7 cells increased to
64% when the cells were treated with BP-PEI-siRNA for 24 h
and 1.0 W cm−2 808 nm irradiation for 10 min. In in vivo
tumor of MCF-7 cells, the combination of BP-PEI-siRNA with
808 nm irradiation had the most potent inhibition of tumor
growth, indicating antisurvivin gene therapy and PTT of BP-
PEI-siRNA had more effective than single therapy. Gold
nanomaterials, the most widely applicable multifunctional
agents in PTT, are usually used in combined therapies based
on their versatile surface chemistry.48,51,58 Using gold nano-
materials, a more effective therapeutic could be achieved. A
gold nanorod was modified by the TAT peptide and thiolated
DNA linker to deliver the sgRNA/Cas9 complex, which
silenced the tumor-associated gene PLK1.59 A significant
inhibiting effect was observed after mild photothermal therapy
combined with gene therapy. A gold nanocage vehicle loading
DOX and siRNA was modified with the AS1411 aptamer for
the combination of tumor-responsive genetic therapy, chemo-
therapy, and photothermal treatment.60 As shown in Figure
2C, a pronounced decrease of VEGF and fewer tumor foci
were seen in the Au-siRNA-PAA-AS1411 group, which was
treated with a combination of tumor-responsive genetic
therapy, chemotherapy, and photothermal treatment. By
designing the ROS-responsive materials, NIR spatially
activated siRNA therapy can be achieved to treat tumor sites
with PDT therapy.61 In normal tissue, siRNA molecules
cannot function due to being trapped and broken down in
endolysosomes. This is caused by the quaternary ammonium
moiety’s unprotonatable property, keeping the siRNA in an
“off” state. However, when NIR irradiation is applied to tumor
tissue, it triggers the release of siRNA from the endolysosomes
and into the cytosol. This is achieved through the cleavage of
TK bonds and the photochemical internalization effect,
resulting in the “on” state of siRNA activity. By inhibiting
the glutathione peroxidase 4 gene, siRNA enhances the
accumulation of ROS, which synergizes with Ce6 photo-
dynamic therapy to provide effective antitumor activity in vivo.
This is achieved through ROS-sensitive cationic nanocarriers,
which provide a feasible and controlled strategy for delivering
siRNA therapy with synergistic drug effects specific to tumors.
The enhanced mechanism of combining gene therapy with
phototherapy can be divided in the following ways.
3.2.2. Increasing the Efficacy of Phototherapy. The
increased thermotolerance induced mainly by the upregulation
of heat shock proteins (HSPs) is the main reason for the failure
of PTT.62 HSPs can act as ATP-mediated molecular
chaperones to correct proteins’ misfolding, thereby keeping
their activities under hyperthermia.62,63 For thermotolerance in
ACS Mater. Au XXXX, XXX, XXX−XXX
ACS Materials Au pubs.acs.org/materialsau
many cancer cells, HSP70 and HSP90 are crucial proteins.63,64
Applying siRNA against the HSP family can improve cells’
sensitivity to PTT and enhance the efficiency of PTT.58,63,65
Bcl-2-associated athanogene domain 3 (BAG3), acting as a
molecular cochaperone of the HSP family, including HSP70, is
also an antiapoptotic protein.66−68 The evidence showed that
the BAG3 overexpression was closely related to the tolerance
of PTT, radiation therapy, and chemotherapy.69 The
combination of BAG3-siRNA and gold nanorods (GNRs) in
gene and photothermal therapy was studied. Treating GNRs-
mediated PTT can significantly increase the expression of
BAG3 and HSP families (HSP27, HSP60, HSP70, and
HSP90) in Cal-27 cells, which indicates the PTT-induced
heat shock response. However, applying BAG3-siRNA could
significantly inhibit the heat shock response, which could
strengthen cells’ sensitivity to PTT. The GNRs-siBAG3
complex showed a stronger BAG3 silencing capability than
lipofectamine 2000-siBAG3 and a higher efficiency of PTT
than GNRs themselves, which completely suppressed the
growth of Cal-27 tumors in vivo.
The generation of HSPs after PTT relies on an abundant
ATP-meditated energy metabolism.70 Therefore, weakening
the energy metabolism seems to be a potential solution to
downregulate HSPs expression during PTT.65 Pyruvate kinase
M2 (PKM2) is involved in ATP production in cancer cells.71
In recent work, a combined strategy of gene therapy and PTT
was reported.64 Simply put, the spherical dendrimer-polypep-
tide (DPP) was synthesized through the PAMAM-mediated
ring-opening polymerization of N-carboxy anhydride. Then,
DPP was used to load ICG and PKM2-siRNA and finally was
coated with human serum albumin. Compared to commercial
transfection reagents (25 kDa PEI and Lipofectamine 2000),
DPP had a dramatically more substantial cellular uptake
capacity and a higher PKM2 silencing efficiency (∼87%) in
MCF-7 cells. Both qPCR and Western blot in vivo and in vitro
showed the same results: the inhibition of PKM2 expression
could remarkably inhibit the production of HSP70 and HSP90
due to the lack of ATP. The triple synergy of siPKM2-induced
cell starvation, downregulated HSP expression, and PTT
showed surprisingly anticancer efficacy for in vivo MCF-7
xenograft model. The tumors of the combined therapy group
were completely suppressed within the observation period, and
the survival rate of the combined therapy group was
satisfactorily 100% within the observation period of 50 days,
which is significantly higher than those of other groups.
The local PTT inevitably induces the heat damage of
surrounded normal cells, which is likely to cause cancer cell
metastasis and local inflammation.72−74 Low-temperature PTT
seems an excellent choice to avoid the side effects because it
relies on low-temperature heat-induced cellular apopto-
sis.72,75,76 The efficacy of low-temperature PTT to solid
tumors is still undesirable due to increased thermotolerance
induced by upregulated expression of intracellular HSPs.77,78 A
general strategy was investigated for combining low-temper-
ature PTT and gene therapy against HSP70.63 First, a siRNA-
bearing self-assembled nucleic acid nanogel based on nucleic
acid hybridization between DNA-grafted polycaprolactone
(DNA-g-PCL) and siRNA linker was developed.79 The
siRNA-bearing nucleic acid nanogel was coated with polydop-
amine and surface PEGylated, named PP-NG. The in vivo
efficacy of PP-NG-siHSP70 combined with low-temperature
PTT was then evaluated on HeLa tumor xenografts.
G https://doi.org/10.1021/acsmaterialsau.3c00035
Review
Furthermore, the Hela tumor growth was almost completely
suppressed under the treatment of PP-NG-siHSP70 (laser),
and more surprisingly, two-thirds of the tumors completely
disappeared. Moreover, PP-NG-siHSP70 plus laser could
induce the most effective inhibition of HSP70 expression
and increase the level of caspase-3 expression. This triple-shield
complex PP-NG-siHSP70 showed the capability of strongly
ablating solid tumors under mild conditions. Further, this gene
delivery system could be expanded to other combined
therapies such as gene/photothermal/immune combined
therapy.63
PDT could be strengthened by downregulating the selected
gene. Recently, chemophotodynamic combination therapy has
become an ideal strategy. However, tumor cells treated by
PDT will induce an abnormal increase in angiogenesis for
generating reactive oxygen species (ROS). A complex
composed of two compartments was developed to suppress
tumor angiogenesis.80 The first compartment is nanoparticle
loading DOX and siRNA silencing the VEGF gene, while the
other part is a Ce6-encapsulating microbubble. Three therapies
showed significantly lower tumor growth, indicating that
overexpression of VEGF was successfully prevented, and
further angiogenesis was restrained (Figure 2D and E).
Survivin, an antiapoptosis protein, was overexpressed in
tumor cells after treatment, thus leading to therapeutic
resistance in tumors.81,82 To downregulate the survival level
in cancer cells, Jin et al. designed an upconversion nanoplat-
form for enhanced PDT.83 They used upconversion nano-
particles adsorbing a long single-stranded DNA (ssDNA) with
multiple copies of the aptamer (AS1411) and DNAzyme to
specifically target nucleolin overexpressed on cancer cells
specifically. The AS1411 aptamer could encapsulate the
photosensitizer 5,10,15,20-tetrakis (1-methylpyridinium-4-yl)
porphyrin (TMPyP4) as well as recognize the nucleolin
overexpressed in tumor. The upconversion nanoplatform could
trigger PDT not only under NIR light but also with high
recognition and loading capacity. The combined therapy
dramatically silences surviving gene expression, hence enhanc-
ing the efficiency of PDT.
3.2.3. Inhibiting Growth and Metastasis of Tumor
Cells to Assist PDT/PTT in Treatment of Tumors. Cancer
metastasis is the leading cause of tumor recurrence. Though
some single therapies like PTT, 84 PDT, MHT, and
sonodynamic therapy (SDT) have been considered promising
therapies because of their stable spatiotemporal control, they
are designed as localized anticancer treatments that are
powerless against the cancers that transfer or survive after
treatment.40,43
Human telomerase reverses transcriptase (hTERT) is
closely related to cancer metastasis and growth.85 A dual-
functionalized BP NS modified with poly(ethylene glycol)
(PEG) and PEI (PPBP) was designed to deliver hTERT
siRNA for gene/photodynamic/photothermal combination
therapy.86 Through the detection of the degradation of
PPBP under different conditions, it is found that PPBP
degrades more significantly under a combination treatment of
H2O2 and PBS of pH 5.0, compared with a single treatment of
H2O2 or PBS of pH 5.0, indicating PPBP can be specifically
degraded in the microenvironment of acidic lysosome and high
photoinduced ROS. si-hTERT synergizes with PDT and PTT
on Hela cells. Moreover, results showed that the introduction
of si-hTERT can effectively cooperate with PTT/PDT therapy
ACS Mater. Au XXXX, XXX, XXX−XXX
ACS Materials Au pubs.acs.org/materialsau Review

Figure 3. Synergistic therapeutic effect of MHT and gene therapy. (A) Scheme of magnetic hyperthermia therapy and gene therapy. (B) The shape
of differently shaped M-MSNs, plots of inverse transverse relaxation time (1/T2) versus Fe concentration of the M-MSNs, and photographs of
HepG2 tumor-bearing mice in each group and MR images of mice from different groups at day 7 and 14 after treatment (tumors marked by the
white arrow). Reprinted with permission from ref 104. Copyright 2018, Elsevier B.V. (C) The effects of let-7a gene and FACS analysis of
combination-treated cells compared to controls. Reprinted with permission from ref 109. Copyright 2014, Wiley-VCH. (D) RT-PCR
demonstrating the successful synthesis of the sTRAIL-EGFP plasmid transfected into A2780 ovarian cancer cells, and tumor volume was followed
over 2 weeks. The size of the tumors decreased significantly when treated with the engineered AD-MSCs. Reprinted with permission from ref 110.
Copyright 2016, Elsevier B.V.

to inhibit the metastasis and growth of A549 cells in vivo As described above, the TRAIL gene has been used wildly in
through downregulating hTERT (Figure 2F and 2G).86 gene therapy. Single gene therapy will lead to uncontrollable
H https://doi.org/10.1021/acsmaterialsau.3c00035
ACS Mater. Au XXXX, XXX, XXX−XXX
ACS Materials Au pubs.acs.org/materialsau
expression of TRAIL in vivo, which phototherapy could solve
for its property of controlled drug release. Meanwhile, the
recurrence of tumor cells after PTT could be killed by TRAIL
treatment. For example, Qiao et al. developed semiconducting
nanoparticles decorated with caged TRAIL-expressing plasmid
under HSP70 protomer named SPN@HSP70-TRAIL-GFP-
(SPNHT).87 SPNHT has great potential to induce cell
apoptosis in vitro and in vivo with a laser, indicating that
combined therapy could effectively restrain tumor recurrence
after PTT treatment.
3.2.4. Protecting Normal Cells and Reducing the
Toxicity of PTT/PDT. The traditional treatment and some
other new therapies like PTT and PDT have high antitumor
efficiency, but at the same time, they are likely to be harmful to
normal healthy cells.88 As two significant photo sources are
used in phototherapy, short-wavelength UV/visible light
cannot penetrate deeply into tissue. At the same time, high-
power-density near-infrared (NIR) light can cause undesired
heat damage. To solve this problem, a ROS-degradable
polycation was designed to codeliver a photosensitizer (PS)
and gene drug, which uses far-red light (661 nm) at low optical
power density.89 The polycation could release the P53 gene
under exposure to far-red light and work together with ROS to
induce the death of cells.
In most cases, HSP can be an unfavorable factor during
phototherapy. However, utilizing this property can also be an
excellent way to take control of gene expression. A multifunc-
tional nanocomposite named PBDTQ, which loads a plasmid
inserted CD-TK double suicide gene with HSP70 promoter,
could achieve heat-controlled gene expression.90 Furthermore,
the suicide gene expression of cytosine deaminase (CD) and
herpes simplex virus type-I thymidine kinase (TK) could turn
prodrugs, 5-fluorocytosine (5-FC) and ganciclovir (GCV),
loaded in PBDTQ into their cytotoxic forms of 5-fluorouracil
(5-Fu) and ganciclovir-triphosphate (GCV-TP) to minimize
collateral damage and nontargeted side effect.
In summary, gene therapy is an effective complement to
phototherapy as it enhances therapeutic efficacy and reduces
side effects. For metastatic cancers, a single PTT or PDT may
not be effective. Gene therapy, specifically the use of a
therapeutic nucleic acid such as siRNA, has been widely used
to combine with PTT/PDT to gain a synergistic therapeutic
effect. The synergistic effect of gene therapy and PTT is
achieved in two ways: either they kill tumor cells together, or
gene therapy can improve the sensitivity of tumor cells to PTT,
such as by inhibiting the expression of heat shock proteins.
The inhibition of tumor growth and metastasis is also possible
with the combination of gene therapy and phototherapy. To
reduce the toxicity of phototherapy, far-red light at low optical
power density can be used, and gene expression can be
controlled by utilizing the properties of heat shock proteins.
3.3. Combined Gene Therapy with Magnetic
Hyperthermia Therapy/Magnetothermal Therapy
Magnetic hyperthermia therapy (MHT), another kind of
promising hyperthermia therapy for localized treatment of
solid cancer, utilizes an external alternating magnetic field
(AMF) to selectively heat magnetic nanoparticles (MNPs or
MNs), inducing tumor cell death/apoptosis.91 Magnetic
particle imaging (MPI) can help track the MNPs’ delivery
and carefully control the release of drugs.92 Due to its safety,
high efficacy, and excellent tissue penetration, MHT has
entered the clinical trial stage (ClinicalTrials.gov Identifier:
I https://doi.org/10.1021/acsmaterialsau.3c00035
Review
NCT02033447). In order to be clinically applicable, the
temperature of MHT (43−46 °C) and the accumulation of
MNPs in tissue must be strictly controlled.93 However, since
the frequency f and field amplitude H of AMF (f × H < 5 ×
109 Am−1s−1) is strictly limited by technology and biomedical
reasons and the thermal conversion efficiency of the existing
MNPs is unable to meet the satisfaction, the therapeutic
efficacy of single MHT is still limited and unsatisfactory.94−96
Hence, many combined strategies are developed to enhance
the efficacy of MHT, such as PTT/MHT, PDT/MHT,
chemotherapy/MHT, gene therapy/MHT, and so on.94 In
order to achieve codelivery of heat magnetic nanoparticles and
gene drugs, there are three main strategies as following:
delivering MNPs and therapeutic oligonucleotide separately;
modifying the surface of MNPs with gene vectors like cationic
polymers which are able to load negative oligonucleotide;
using MNPs themselves to load pDNA/polymer complex.97−99
It was reported that a mild multiple MHT-mediated TRAIL
release system was a good solution for TRAIL-resistant cancer
cells. A biodegradable TRAIL/SPION nanocomplex hydrogel
(T/S-NH) designed for combined therapy with MHT and
simultaneously MHT-induced TRAIL release.100
3.3.1. Synergistic Therapeutic Effect of MHT and
Gene Therapy. The unique magnetism gives MNPs
enhanced cellular internalization, controllable tumor accumu-
lation under external magnetic fields, and control of target gene
expression under AMF (Figure 3A). Through multiple surface
modifications of viral or nonviral vectors, MNPs can load
therapeutic nucleotides to achieve the combined gene/
magnetic hyperthermia therapy.101 In the earliest examples of
the combined therapy, they injected 20 μg of pGadTNF into
tumors by a lipofection method for TNF-α gene therapy and,
after 1 day, injected magnetite cationic liposomes into tumors
for MHT under AMF (118 kHz and 30.6 kA/m) for 30 min.
The hyperthermia (46 °C) and overexpression of TNF-α (3-
fold vs control group) resulted in potent inhibition of U251-SP
tumor growth (0.5 ± 0.4 cm3 on day 32), which was
significantly smaller than that of single MHT (2.0 ± 1.1 cm3
on day 32) or TNF-α gene therapy (2.9 ± 1.1 cm3 on day
32).98
Among the numerous gene therapy clinical trials, the herpes
simplex virus thymidine kinase (HSV-TK) gene is the best-
characterized suicide gene for tumor cell killing. The HSV-TK
can make nontoxic ganciclovir monophosphate into toxic
ganciclovir triphosphate, leading to a bystander effect.102 Here
25 kDa PEI-Fe3O4 MNs were developed to deliver the HSV-
TK suicide gene induced by an α-fetoprotein promoter and
hypoxia enhancement for the combined gene/magnetic
hyperthermia therapy of hepatocellular carcinoma (HCC).
They found a synergistic therapeutic effect of the combined
therapy.103 Sphere-like and rod-like shape-controlled magnetic
mesoporous silica nanoparticles (M-MSNs) were developed to
integrate gene therapy, MHT, and magnetic resonance imaging
together.104 Modified with PEG-g-PLL, the positively charged
M-MSNs were used to load ganciclovir and the HSV-TK
plasmid. Both shapes of M-MSNs with ganciclovir and pTK
demonstrated a highly effective combined suicide gene/
magnetic hyperthermia therapy of HCC (Figure 3B).
Compared to sphere-like M-MSNs, rod-like M-MSNs showed
higher magnetic targeting and combined therapeutic perform-
ance (90% inhibition in vivo). Gene therapy could enhance the
effect of MHT in several ways, as described below.
ACS Mater. Au XXXX, XXX, XXX−XXX
ACS Materials Au pubs.acs.org/materialsau
3.3.2. Improving the Sensitivity of Tumor Cells to
MHT. Gene therapy, as an exciting tumor treatment, can
improve the sensitivity of tumor cells to hyperthermia induced
by MHT by regulating heat resistance-related genes like HSPs.
The inhibition of HSPs expression makes it possible for a
practical therapeutic effect of magnetic nanoparticle complex
under mild heating or low concentration, which can decrease
the risk of inflammation and damage to surrounding normal
cells.105 In Court et al.’s study, they detected the gene
expression of ovarian cancer cell line HeyA8 cells after
treatment of MHT (43 °C, 30 min) through microarray
analysis and found that the top 20 upregulated genes contained
HSP70 (HSPA6, HSPA7, HSPA4L) and BAG3.106 Then, they
investigated the combination of HSPA6-siRNA and MHT to
treat ovarian cancer cells in vitro and in vivo. Four kinds of
anti-HSPA6 siRNA were selected to inhibit HSPA6 expression
in ovarian cancer cells (A27880 cP20 and HeyA8), and all four
HSPA6-siRNA-showed a noticeable improvement to CMDx-
IO based MHT in vitro. The similar positive impact of
HSPA6-siRNA on MHT was verified in an in vivo
subcutaneous ovarian tumor model.
Besides siRNA therapy, micro-RNAs are reported to
combine with MHT against HSP. Compared to the gene
therapy based on siRNA, which only modulates single HSP-
related targets, the gene therapy based on miRNA can
modulate multiple targets on the same or similar pathways,
indicating a more significant and broader effect.107 Lethal-7a
miRNA (let-7a), a tumor suppressor, is downregulated in
several human cancers, including brain cancers. Let-7a is
associated with DNA repair and cell survival mechanisms by
targeting factors such as RAS, MYC, and cyclin D, also related
to downstream regulation of the HSP family.108 The combined
therapy of miRNA gene therapy with ZnFe2O4-based MHT
was developed.109 The negatively charged ZnFe2O4 MNPs
were modified with 10 kDa branched PEI to load let-7a
miRNA through a two-step layer-by-layer process. To evaluate
the therapeutic enhancement of let-7a to MHT, U87-
EGFRvIII cells were treated with let-7a, magnetic hyper-
thermia, or the combination. The combined treatment group
exhibited lower cell viability (34%) than the single treatment
groups (let-7a treatment 68.9%, magnetic hyperthermia
63.14%). A mechanistic study of combined therapy showed
the upregulation of caspase-3 and downregulation of PI3K,
HSPs (HSP27, HSP70, HSP72, and HSP90), and let-7a targets
(RAS, MYC, and IGF1R), which suggested that let-7a could
enhance the MHT to glioblastoma multiforme through
inducing cell apoptosis and increasing the sensitivity of cells
to magnetic hyperthermia (Figure 3C). The MNPs can be
further modified with targeting ligands or PEG to improve the
targeting and biocompatibility.
Nevertheless, due to the limitations of magnetic materials
and gene vectors, the combination of gene therapy and MHT
still receives less attention. The development of the combined
therapy should consider the rational combination of MNs,
surface decorations, nucleic acid types, and genes. Recently,
magnetic hyperthermia-mediated gene therapy has been
receiving increased attention. In this gene therapy, MNs
enhance tumor accumulation, therapeutic gene release, and
magnetic hyperthermia-activated gene expression.101 For
example, a novel mild magnetic hyperthermia-activated stem
cell-based gene therapy was developed to heal ovarian cancer.
Here, 25 kDa PEI-modified magnetic core−shell nanoparticles
composd a ZnFe2O4 core and mesoporous silica shell to
J https://doi.org/10.1021/acsmaterialsau.3c00035
Review
deliver and activate heat-inducible TRAIL plasmid with HSP70
B’ promoter in mesenchymal stem cells.110 MCNPs can deliver
a heat-inducible plasmid encoding TRAIL and activate TRAIL
secretion in engineered AD-MSCs with mild magnetic
hyperthermia. Quantifying luminescence intensity shows that
the engineered AD-MSCs are significantly better than
treatment with a single dose of recombinant TRAIL (Figure
3D).
In short, gene therapy can improve the sensitivity of tumor
cells to hyperthermia induced by MHT by regulating heat
resistance-related genes like HSPs. The inhibition of HSPs
expression makes it possible for a practical therapeutic effect of
magnetic nanoparticle complexes under mild heating or low
concentration, which can decrease the risk of inflammation and
damage to surrounding normal cells. Various studies have
reported that combined therapy has shown significant and
broader effects compared to single therapy. Meanwhile, the
development of combined therapy should consider the rational
combination of MNs, surface decorations, nucleic acid types,
and genes.
3.4. Combined Gene Therapy with Immune therapy
The immune system is the key to detecting and eradicating
cancer cells in vivo, involving a series of complex mechanisms.
On the one hand, it can effectively prevent the occurrence and
development of malignant tumors. However, on the other
hand, it will also promote the selection of immune-tolerant
tumor cells. After three stages of tumor development
(elimination, equilibrium, and escape), tumors in the body
have evolved into tumor cells that can evade the immune
response, resulting in uncontrollable cancer in the body.111
The immune system has been considered to be a key point for
cancer healing and detection. Immune cells can be engaged in
a battle with cancer cells. Other cases can promote the
metastasis and recurrence of tumors after three phases of
eliminating, balancing, and escaping tumor cells, which involve
complex mechanisms. The mechanisms for the escape of
cancer cells from the immune system have been widely studied,
including the change of immune checkpoint proteins like PD-
1, and CTLA-4, manipulation of cytokine expression, and
alteration of antigens.111,112 Newer therapies combined with
gene therapy and immunotherapy have flourished in recent
years and made encouraging progress, including OV therapy,
adoptive immunotherapy, and tumor vaccine. Besides, certain
materials exhibit immunomodulatory effects that can poten-
tially synergize with gene therapy drugs to enhance their
efficacy.113
3.4.1. Synergistic Therapeutic Effect of Immunother-
apy and Gene therapy. Immunotherapy is a promising
therapeutic that has been investigated widely in recent years.
The main types of cellular immunotherapy include CAR T,
TCR-transduced T cells (TCR-T), tumor-infiltrating lympho-
cytes (TIL), and natural killer (NK) cells. CAR T therapy is a
hot field in clinical practice. Five cell therapies approved by the
U.S. Food and Drug Administration (FDA) come from this
field. Checkpoint inhibitors (ICI) have revolutionized cancer
treatment and have become the standard of care for many
cancer patients. Although immunotherapy has been applied in
terminal cancer, it is greatly limited by an immunosuppressive
tumor microenvironment for low pH, hypoxia, etc. Gene
therapy could steadily activate immune cells in the tumor
microenvironment or enhance the vaccine effect, thus gaining
the strengthening therapeutic effect (Figure 4A).
ACS Mater. Au XXXX, XXX, XXX−XXX
ACS Materials Au pubs.acs.org/materialsau Review

Figure 4. Synergistic therapeutic effect of immunotherapy and gene therapy. (A) Scheme of immunotherapy and gene therapy. (B) Western
blotting of OVA proteins in RAW264.7 and DC2.4, showing the expression of OVA in GLP-O and GLP-RONP groups and H&E images of lung
tissues collected on day 40, showing that the GLP-RO Gel could efficiently prevent the formation of metastasis. Reprinted with permission from ref
118. Copyright 2021, American Chemical Society. (C) The polarization states of macrophages stimulated with supernatants derived from
transfected B16F10 cells were assessed by FCM. The PD-L1 expression levels in tumors subjected to different treatments were assessed by FCM.
Codelivery of CCL19 pDNA and BMS-1 in RGD-DMA nanoparticles reshaped the TME and significantly inhibited the abdominal dissemination
of cancer. Reprinted with permission from ref 121. Copyright 2021, Wiley-VCH. (D) Immunohistochemical images showing the expression of PD-
L1 (brown) in CT26 tumor tissue. Red arrows indicate PD-L1 protein. Western blot analysis of PD-L1 expression in CT26 tumor tissue collected
from mice in the indicated treatment groups and Statistical analysis of the number of TUNEL+ cells per field for B16 and CT26 tumor. Scale bar =
50 μm. Reprinted with permission from ref 123. Copyright 2020, American Chemical Society.

3.4.2. Using an Oncolytic Virus to Combat Tumors.
Immunity therapy is usually combined with oncolytic virus
therapy because oncolytic virus (OVs) could induce
immunogenic cell death, release tumor-associated antigens
(TAAs), disrupt immunosuppression within tumors, and
promote antitumor immune responses.114 The Phase I study
of Delta-24-RGD oncolytic adenovirus showed a significant
antitumor efficiency because of a direct oncolytic effect and a
following antitumor immune response.115 The oncolytic virus/
immune combined therapies of Delta-24-RGD and anti-PD-1
antibodies are undergoing Phase II clinical trials (Clinical-
Trials.gov identifier: NCT02798406). Through modifying
antitumor cytokine or antiviral cytokine genes, oncolytic
virus therapy combined with gene therapy can improve the
relative dismal efficacy of a single oncolytic virus agent or
reduce the virus-induced toxicity. Amgen’s T-Vec (talimogene
laherparepvec), the first approved oncolytic virus by FDA, was
modified with the immunostimulatory cytokine granulocyte-
macrophage colony-stimulating factor (GM-CSF) gene and
gained a highly therapeutic effect in melanoma patients with a
durable response rate of 16% in advanced melanoma patients
K https://doi.org/10.1021/acsmaterialsau.3c00035
to 2% in the control group.116 Stephen Russell at Mayo Clinic
developed a recombinant oncolytic vesicular stomatitis virus
(VSV) expressing the antiviral cytokine interferon-β to protect
noncancerous cells from being killed.116,117 However, the
immune response induced by OVs or the cytokines expressed
by OVs is likely to be systemic, increasing the risk of OVs’
toxicity. A modified oncolytic adenovirus Delta-24-RGDOX
was developed to express an immune costimulatory OX40L
(OX40 ligand) which can bind a unique costimulatory OX40
on the T cell membrane and help T cells to recognize TAAs.
The treatment induced complete regression in the long term
surviving mice treated with the combination. Combining OX,
immune therapy, and the unique gene OX40L gave Delta-24-
RGDOX a specific immune response to cancer cells,
significantly reducing the potential immunotoxicity of OVs.114
3.4.3. Nucleic Acid-Based Vaccine. Nanovaccine is a
new therapeutic that can activate the immune system by
presenting an antigen to lymphocytes, thus killing tumor cells.
The former vaccine usually contains an antigen alone or with
an adjuvant, which cannot be recognized by lymphocytes
effectively. With nanocarriers, vaccines could effectively target
ACS Mater. Au XXXX, XXX, XXX−XXX
ACS Materials Au pubs.acs.org/materialsau
specific organs and transform them into cells. The nucleic acid
vaccine, especially the mRNA vaccine, has become one of the
most potent tools against diseases such as covid-19 and cancer
because of its long-term effect and safety. Once mRNA enters
the cell, it can express specific proteins continuously, thus
making immune cells produce antibodies. A nanovaccine
formed with ovalbumin mRNA and adjuvants (R848) was
loaded in injectable hydrogel to gain the enhanced effect.118
The nanovaccine could generate an OVA 30 days after one
injection and target lymph nodes. Results show that generating
tumor antigen antibodies in the serum could effectively prevent
metastasis formation (Figure 4B). To encapsulate individual
tumor mRNA into cationic lipid nanoparticles, the RNA-NPs
could activate systemic and intratumoral myeloid cells.119
Positive charge lipid nanoparticles encapsulating untargeted
tumor RNA can activate the response to immunotherapy.
Codelivery of anti-PD-L1 mAbs and RNA-NPs could increase
peripheral/intratumoral PD-1+ CD8+ cells and elicits anti-
tumor activity in “cold” tumor models resistant to ICI
monotherapy. Nanovaccine could also work together with
implantable materials to gain an enhanced efficacy. A blood
clot scaffold loading liposome vaccine and siRNAs was
developed to recruit and induce the maturation of DCs and
reduce immunosuppressive signals in mature DCs120. By
loading PD-L1/TIM-3 siRNA, the scaffold could reduce the
expression of immunosuppressive molecules in DCs thus
stimulating a strong immune response against tumors. It can
also treat different tumors by changing tumor antigens. The
system can be optimized by encapsulating components into the
same nanoparticle in the future.120
3.4.4. Changing Tumor Microenvironment from Cold
to Hot. Besides using nucleic acids to stimulate the immune
system against the tumor, utilizing ICI, cytokines, and other
immunostimulatory molecules could also heal cancer by the
immune system. However, the immunosuppressive tumor
microenvironment significantly limits these traditional immu-
notherapies for their low pH, hypoxia, etc. By transferring a
specific gene or interfering with gene expression, the tumor
microenvironment could be changed, enhancing the effect of
immunotherapy. The nanoparticles named RGD-DMA
encapsulating CCL19-encoding plasmid DNA (CCL19
pDNA) and immune checkpoint ligand PD-L1 inhibitor
(BMS-1) were developed to overcome the immunosuppressive
TME and gain enhanced efficacy.121 CCL-19 could stimulate T
cell proliferation, thus increasing IFN-γ secretion in the TME,
while BMS-1 could inhibit the expression of PD-L1 more
efficiently. Nanoparticles could reshape the TME and inhibit
cancer’s abdominal dissemination (Figure 4C). Tumor-
targeted lipid-dendrimer-calcium-phosphate (TT-LDCP)
nanoparticles with thymine functionalization were designed
to gain the enhanced gene delivery capacity and adjuvant
immune properties by activating the stimulator of interferon
genes (STING) pathway.122 TT-LDCP NPs could deliver
siRNA against PD-L1 and plasmid DNA, which could encode
IL-2 in tumors, activate CD8+ T cells, augment the efficacy of
cancer immunotherapy, and suppress tumor progression. A
bacterial outer membrane vesicle-based immunotherapeutic
agent has been studied for tumor immunotherapy.123 It
activates the immune system and disrupts the PD-L1-mediated
immunosuppressive tumor microenvironment. PD1-bearing
OMVs significantly enhance the antitumor immune response.
OMV-PD1 stimulates the tumor infiltration of immune cells
and a systemic antitumor immune response. The strong
L https://doi.org/10.1021/acsmaterialsau.3c00035
Review
accumulation of the OMV-PD1 at the tumor and the vesicles’
efficient binding to PD-L1 on tumor cells deplete tumor cell
PD-L1 and ultimately block the PD1/PD-L1 inhibitory axis.
The combination of immune activation and blockage of PD1/
PD-L1 leads to a significant reduction in tumor growth in
mouse melanoma and colorectal cancer models. The
engineered bacterial extracellular vesicle-based system provides
an exciting opportunity to further develop tumor immuno-
therapy strategies (Figure 4D).
In summary, combining immunotherapy with gene therapy
can enhance the therapeutic effect of cancer treatment.
Oncolytic virus therapy combined with gene therapy has also
shown promise in disrupting immunosuppression within
tumors and promoting antitumor immune responses. Nucleic
acid-based vaccines, such as mRNA vaccines, have become
powerful tools against cancer due to their long-term effect and
safety. Changing the tumor microenvironment by gene therapy
can also enhance the effectiveness of immunotherapy.
4. CONCLUSION AND FUTURE PERSPECTIVE
Combined gene therapy has many advantages that a single
therapeutic cannot achieve, such as preventing tumor meta-
stasis and recurrence, enhanced therapeutic efficiency, etc. By
encapsulating drugs into nanocarriers, the delivery system
could achieve targeting ability, lysosome escape, and other
functions. Understanding the mechanisms of synergistic
therapeutic effects is crucial for designing therapeutic schemes
and the forms of nanocarriers. However, there are still several
barriers to combining gene therapy with nanoplatform. First,
the metabolism and toxicity of different nanocarriers must be
researched systematically. Although many nanocarriers have
been researched, a smaller number of carriers have been
approved by the FDA. Before clinical use, it is crucial to
thoroughly investigate the interaction between various nano-
materials and organisms as well as determine whether these
materials can be degraded in vivo. These factors must be
clearly understood to ensure safe and effective use. Second, the
efficiency of gene transformations still has much room for
improvement. How to synthesize a nanocarrier that could
deliver specific nucleic acids into cells with no apparent toxicity
is a big challenge. Besides, the transform of gene drugs is
critical for evaluation of dosage of administration and the
proportion of drugs contained in the delivery system. Third,
the long-term effects of combined therapy should be
investigated, which should observe if gene drugs have other
effects on normal organs and if it is suitable for long-term use.
Gene therapy is a highly sought-after treatment option due to
its ability to provide a cure with just one dose or treatment,
unlike other therapies. However, to ensure optimal therapeutic
results, it is crucial to assess the appropriate interval of time for
combined therapy prior to clinical use.
■ AUTHOR INFORMATION
Corresponding Author
Ying Liu − CAS Key Laboratory for Biomedical Effects of
Nanomaterials and Nanosafety and CAS Center for
Excellence in Nanoscience, National Center for Nanoscience
and Technology of China, Beijing 100190, P.R. China;
orcid.org/0000-0002-6540-0473; Phone: +8610
82545526; Email: liuy@nanoctr.cn
ACS Mater. Au XXXX, XXX, XXX−XXX
ACS Materials Au
Authors
Jinhui Lin − CAS Key Laboratory for Biomedical Effects of
Nanomaterials and Nanosafety and CAS Center for
Excellence in Nanoscience, National Center for Nanoscience
and Technology of China, Beijing 100190, P.R. China;
University of Chinese Academy of Sciences, Beijing 100049,
P.R. China
Xinlian Wang − CAS Key Laboratory for Biomedical Effects of
Nanomaterials and Nanosafety and CAS Center for
Excellence in Nanoscience, National Center for Nanoscience
and Technology of China, Beijing 100190, P.R. China;
University of Chinese Academy of Sciences, Beijing 100049,
P.R. China
Dongqi Ni − CAS Key Laboratory for Biomedical Effects of
Nanomaterials and Nanosafety and CAS Center for
Excellence in Nanoscience, National Center for Nanoscience
and Technology of China, Beijing 100190, P.R. China;
University of Chinese Academy of Sciences, Beijing 100049,
P.R. China
Yandong Chen − CAS Key Laboratory for Biomedical Effects
of Nanomaterials and Nanosafety and CAS Center for
Excellence in Nanoscience, National Center for Nanoscience
and Technology of China, Beijing 100190, P.R. China
Chunying Chen − CAS Key Laboratory for Biomedical Effects
of Nanomaterials and Nanosafety and CAS Center for
Excellence in Nanoscience, National Center for Nanoscience
and Technology of China, Beijing 100190, P.R. China;
University of Chinese Academy of Sciences, Beijing 100049,
P.R. China; orcid.org/0000-0002-6027-0315
Complete contact information is available at:
https://pubs.acs.org/10.1021/acsmaterialsau.3c00035
Author Contributions
CRediT: Jinhui Lin formal analysis (equal), validation (equal),
writing-original draft (lead); Xinlian Wang resources (support-
ing); Dongqi Ni writing-original draft (supporting); Yandong
Chen resources (supporting), funding acquisition (support-
ing); Chunying Chen funding acquisition (lead), writing-
review & editing (equal); Ying Liu conceptualization (lead),
funding acquisition (equal), writing-review & editing (lead).
Notes
The authors declare no competing financial interest.
■ ACKNOWLEDGMENTS
This work was financially supported by the National Natural
Science Foundation of China (31971318, 22027810,
32101091) and China Postdoctoral Science Foundation
(2021M690043).
■ REFERENCES
(1) Siegel, R. L.; Miller, K. D.; Fuchs, H. E.; Jemal, A. Cancer
statistics, 2022. CA: A Cancer Journal for Clinicians 2022, 72 (1), 7−
33.
(2) Miller, K. D.; Nogueira, L.; Mariotto, A. B.; Rowland, J. H.;
Yabroff, K. R.; Alfano, C. M.; Jemal, A.; Kramer, J. L.; Siegel, R. L.
Cancer treatment and survivorship statistics, 2019. CA: A Cancer
Journal for Clinicians 2019, 69 (5), 363−385.
(3) Pan, L.; Liu, J.; Shi, J. Cancer cell nucleus-targeting
nanocomposites for advanced tumor therapeutics. Chem. Soc. Rev.
2018, 47 (18), 6930−6946.
pubs.acs.org/materialsau Review
(4) Liu, Y.; Bhattarai, P.; Dai, Z.; Chen, X. Photothermal therapy
and photoacoustic imaging via nanotheranostics in fighting cancer.
Chem. Soc. Rev. 2019, 48 (7), 2053−2108.
(5) Ni, D.; Lin, J.; Zhang, N.; Li, S.; Xue, Y.; Wang, Z.; Liu, Q.; Liu,
K.; Zhang, H.; Zhao, Y.; Chen, C.; Liu, Y. Combinational application
of metal-organic frameworks-based nanozyme and nucleic acid
delivery in cancer therapy. WIREs Nanomedicine and Nanobiotechnol-
ogy 2022, 14 (3), No. e1773.
(6) Huang, J.; Xiao, Z.; Chen, G.; Li, T.; Peng, Y.; Shuai, X. A pH-
sensitive nanomedicine incorporating catalase gene and photo-
sensitizer augments photodynamic therapy and activates antitumor
immunity. Nano Today 2022, 43, 101390.
(7) Dunbar, C. E.; High, K. A.; Joung, J. K.; Kohn, D. B.; Ozawa, K.;
Sadelain, M. Gene therapy comes of age. Science 2018, 359 (6372),
No. eaan4672.
(8) D’Aloia, M. M.; Zizzari, I. G.; Sacchetti, B.; Pierelli, L.; Alimandi,
M. CAR-T cells: the long and winding road to solid tumors. Cell
Death & Disease 2018, 9 (3), 282.
(9) Zhao, N.; Yan, L.; Xue, J.; Zhang, K.; Xu, F.-J. Degradable one-
dimensional dextran-iron oxide nanohybrids for MRI-guided syner-
gistic gene/photothermal/magnetolytic therapy. Nano Today 2021,
38, 101118.
(10) Li, Y.; Liu, L.; Ji, W.; Peng, H.; Zhao, R.; Zhang, X. Strategies
and materials of ″SMART″ non-viral vectors: Overcoming the
barriers for brain gene therapy. Nano Today 2020, 35, 101006.
(11) Qiu, N.; Wang, G.; Wang, J.; Zhou, Q.; Guo, M.; Wang, Y.; Hu,
X.; Zhou, H.; Bai, R.; You, M.; Zhang, Z.; Chen, C.; Liu, Y.; Shen, Y.
Tumor-Associated Macrophage and Tumor-Cell Dually Transfecting
Polyplexes for Efficient Interleukin-12 Cancer Gene Therapy. Adv.
Mater. 2021, 33 (2), 2006189.
(12) Song, G.; Cheng, L.; Chao, Y.; Yang, K.; Liu, Z. Emerging
Nanotechnology and Advanced Materials for Cancer Radiation
Therapy. Adv. Mater. 2017, 29 (32), 1700996.
(13) Shen, J.; Zhang, W.; Qi, R.; Mao, Z.-W.; Shen, H. Engineering
functional inorganic-organic hybrid systems: advances in siRNA
therapeutics. Chem. Soc. Rev. 2018, 47 (6), 1969−1995.
(14) Teo, P. Y.; Cheng, W.; Hedrick, J. L.; Yang, Y. Y. Co-delivery of
drugs and plasmid DNA for cancer therapy. Adv. Drug Delivery Rev.
2016, 98, 41−63.
(15) Jiang, D. M.; Gupta, S.; Kitchlu, A.; Meraz-Munoz, A.; North, S.
A.; Alimohamed, N. S.; Blais, N.; Sridhar, S. S. Defining cisplatin
eligibility in patients with muscle-invasive bladder cancer. Nature
Reviews Urology 2021, 18 (2), 104−114.
(16) Liu, S.; Khan, A. R.; Yang, X.; Dong, B.; Ji, J.; Zhai, G. The
reversal of chemotherapy-induced multidrug resistance by nano-
medicine for cancer therapy. J. Controlled Release 2021, 335, 1−20.
(17) Belkahla, H.; Herlem, G.; Picaud, F.; Gharbi, T.; Hémadi, M.;
Ammar, S.; Micheau, O. TRAIL-NP hybrids for cancer therapy: a
review. Nanoscale 2017, 9 (18), 5755−5768.
(18) Zhong, H.-h.; Wang, H.-y.; Li, J.; Huang, Y.-z. TRAIL-based
gene delivery and therapeuticstrategies. Acta Pharmacologica Sinica
2019, 40 (11), 1373−1385.
(19) Xu, F.; Zhong, H.; Chang, Y.; Li, D.; Jin, H.; Zhang, M.; Wang,
H.; Jiang, C.; Shen, Y.; Huang, Y. Targeting death receptors for drug-
resistant cancer therapy: Codelivery of pTRAIL and monensin using
dual-targeting and stimuli-responsive self-assembling nanocomposites.
Biomaterials 2018, 158, 56−73.
(20) Amreddy, N.; Babu, A.; Panneerselvam, J.; Srivastava, A.;
Muralidharan, R.; Chen, A.; Zhao, Y. D.; Munshi, A.; Ramesh, R.
Chemo-biologic combinatorial drug delivery using folate receptor-
targeted dendrimer nanoparticles for lung cancer treatment. Nano-
medicine: Nanotechnology, Biology and Medicine 2018, 14 (2), 373−
384.
(21) Qiu, L.; Chen, T.; Ocsoy, I.; Yasun, E.; Wu, C.; Zhu, G.; You,
M.; Han, D.; Jiang, J.; Yu, R.; Tan, W. A cell-targeted, size-
photocontrollable, nuclear-uptake nanodrug delivery system for drug-
resistant cancer therapy. Nano Lett. 2015, 15 (1), 457−63.
(22) Shu, J.; Li, X.; Dang, J.; Liu, Y.; Duan, S.; Zhu, R.; Yin, L.;
Chen, Y. Drug Resistance Reversal by Interventing Cancer
M https://doi.org/10.1021/acsmaterialsau.3c00035
ACS Mater. Au XXXX, XXX, XXX−XXX
ACS Materials Au pubs.acs.org/materialsau
Bioenergetics with Spherical Helical Polypeptide-Potented Gene
Silencing. Chemical Engineering Journal 2021, 414 (10), 128545.
(23) Bukowski, K.; Kciuk, M.; Kontek, R. Mechanisms of Multidrug
Resistance in Cancer Chemotherapy. International Journal of
Molecular Sciences 2020, 21 (9), 3233.
(24) Zhang, L.; Mu, C.; Zhang, T.; Yang, D.; Wang, C.; Chen, Q.;
Tang, L.; Fan, L.; Liu, C.; Shen, J.; Li, H. Development of targeted
therapy therapeutics to sensitize triple-negative breast cancer
chemosensitivity utilizing bacteriophage phi29 derived packaging
RNA. J. Nanobiotechnol. 2021, 19 (1), 13.
(25) Jing, X.; Yang, F.; Shao, C.; Wei, K.; Xie, M.; Shen, H.; Shu, Y.
Role of hypoxia in cancer therapy by regulating the tumor
microenvironment. Molecular Cancer 2019, 18 (1), 157.
(26) Xie, Z.; Guo, W.; Guo, N.; Huangfu, M.; Liu, H.; Lin, M.; Xu,
W.; Chen, J.; Wang, T.; Wei, Q.; Han, M.; Gao, J. Targeting tumor
hypoxia with stimulus-responsive nanocarriers in overcoming drug
resistance and monitoring anticancer efficacy. Acta biomaterialia 2018,
71, 351.
(27) Valkenburg, K. C.; de Groot, A. E.; Pienta, K. J. Targeting the
tumour stroma to improve cancer therapy. Nat. Rev. Clin Oncol 2018,
15 (6), 366−381.
(28) Ni, Y.; Zhou, X.; Yang, J.; Shi, H.; Li, H.; Zhao, X.; Ma, X. The
Role of Tumor-Stroma Interactions in Drug Resistance Within
Tumor Microenvironment. Frontiers in Cell and Developmental Biology
2021, 9 (1206), na DOI: 10.3389/fcell.2021.637675.
(29) Larionova, I.; Cherdyntseva, N.; Liu, T.; Patysheva, M.; Rakina,
M.; Kzhyshkowska, J. Interaction of tumor-associated macrophages
and cancer chemotherapy. OncoImmunology 2019, 8 (7),
No. e1596004.
(30) Parayath, N. N.; Gandham, S. K.; Leslie, F.; Amiji, M. M.
Improved anti-tumor efficacy of paclitaxel in combination with
MicroRNA-125b-based tumor-associated macrophage repolarization
in epithelial ovarian cancer - ScienceDirect. Cancer letters 2019, 461,
1−9.
(31) Tcyganov, E.; Mastio, J.; Chen, E.; Gabrilovich, D. I. Plasticity
of myeloid-derived suppressor cells in cancer. Current Opinion in
Immunology 2018, 51, 76−82.
(32) Chen, Y.; Sun, J.; Huang, Y.; Liu, Y.; Liang, L.; Yang, D.; Lu, B.;
Li, S. Targeted codelivery of doxorubicin and IL-36γ expression
plasmid for an optimal chemo-gene combination therapy against
cancer lung metastasis. Nanomedicine: Nanotechnology, Biology and
Medicine 2019, 15, 129−141.
(33) Hwang, R. F.; Moore, T.; Arumugam, T.; Ramachandran, V.;
Amos, K. D.; Rivera, A.; Ji, B.; Evans, D. B.; Logsdon, C. D. Cancer-
associated stromal fibroblasts promote pancreatic tumor progression.
Cancer Res. 2008, 68 (3), 918−26.
(34) Chen, X.; Jia, F.; Li, Y.; Deng, Y.; Huang, Y.; Liu, W.; Jin, Q.; Ji,
J. Nitric oxide-induced stromal depletion for improved nanoparticle
penetration in pancreatic cancer treatment. Biomaterials 2020, 246,
119999.
(35) Han, X.; Li, Y.; Xu, Y.; Zhao, X.; Zhang, Y.; Yang, X.; Wang, Y.;
Zhao, R.; Anderson, G. J.; Zhao, Y.; Nie, G. Reversal of pancreatic
desmoplasia by re-educating stellate cells with a tumour micro-
environment-activated nanosystem. Nat. Commun. 2018, 9 (1), 3390.
(36) Li, Y.; Zhao, Z.; Lin, C.-Y.; Liu, Y.; Staveley-OCarroll, K. F.; Li,
G.; Cheng, K. Silencing PCBP2 normalizes desmoplastic stroma and
improves the antitumor activity of chemotherapy in pancreatic cancer.
Theranostics 2021, 11 (5), 2182−2200.
(37) Wu, F.; Yang, J.; Liu, J.; Wang, Y.; Mu, J.; Zeng, Q.; Deng, S.;
Zhou, H. Signaling pathways in cancer-associated fibroblasts and
targeted therapy for cancer. Signal Transduction and Targeted Therapy
2021, 6 (1), 218.
(38) Saatci, O.; Kaymak, A.; Raza, U.; Ersan, P. G.; Akbulut, O.;
Banister, C. E.; Sikirzhytski, V.; Tokat, U. M.; Aykut, G.; Ansari, S. A.;
Dogan, H. T.; Dogan, M.; Jandaghi, P.; Isik, A.; Gundogdu, F.;
Kosemehmetoglu, K.; Dizdar, O.; Aksoy, S.; Akyol, A.; Uner, A.;
Buckhaults, P. J.; Riazalhosseini, Y.; Sahin, O. Targeting lysyl oxidase
(LOX) overcomes chemotherapy resistance in triple negative breast
cancer. Nat. Commun. 2020, 11 (1), 2416.
N https://doi.org/10.1021/acsmaterialsau.3c00035
Review
(39) Qin, L.; Jiang, S.; He, H.; Ling, G.; Zhang, P. Functional black
phosphorus nanosheets for cancer therapy. J. Controlled Release 2020,
318, 50−66.
(40) Fusco, L.; Gazzi, A.; Peng, G.; Shin, Y.; Vranic, S.; Bedognetti,
D.; Vitale, F.; Yilmazer, A.; Feng, X.; Fadeel, B.; Casiraghi, C.; Delogu,
L. G. Graphene and other 2D materials: a multidisciplinary analysis to
uncover the hidden potential as cancer theranostics. Theranostics
2020, 10 (12), 5435−5488.
(41) Kim, J.; Kim, J.; Jeong, C.; Kim, W. J. Synergistic nanomedicine
by combined gene and photothermal therapy. Adv. Drug Delivery Rev.
2016, 98, 99−112.
(42) Kim, N. Y.; Blake, S.; De, D.; Ouyang, J.; Shi, J.; Kong, N. Two-
Dimensional Nanosheet-Based Photonic Nanomedicine for Com-
bined Gene and Photothermal Therapy. Frontiers in Pharmacology
2020, 10, na DOI: 10.3389/fphar.2019.01573.
(43) Cai, Y.; Ni, D.; Cheng, W.; Ji, C.; Wang, Y.; Müllen, K.; Su, Z.;
Liu, Y.; Chen, C.; Yin, M. Enzyme-Triggered Disassembly of Perylene
Monoimide-based Nanoclusters for Activatable and Deep Photo-
dynamic Therapy. Angew. Chem., Int. Ed. 2020, 59 (33), 14014−
14018.
(44) Chen, W. H.; Luo, G. F.; Zhang, X. Z. Recent Advances in
Subcellular Targeted Cancer Therapy Based on Functional Materials.
Adv. Mater. 2019, 31 (3), No. 1802725.
(45) Nam, J.; Son, S.; Ochyl, L. J.; Kuai, R.; Schwendeman, A.;
Moon, J. J. Chemo-photothermal therapy combination elicits anti-
tumor immunity against advanced metastatic cancer. Nat. Commun.
2018, 9 (1), 1074.
(46) Min, K. H.; Kim, Y. H.; Wang, Z.; Kim, J.; Kim, J. S.; Kim, S.
H.; Kim, K.; Kwon, I. C.; Kiesewetter, D. O.; Chen, X. Engineered
Zn(II)-Dipicolylamine-Gold Nanorod Provides Effective Prostate
Cancer Treatment by Combining siRNA Delivery and Photothermal
Therapy. Theranostics 2017, 7 (17), 4240−4254.
(47) Zhang, Y.; Ren, K.; Zhang, X.; Chao, Z.; Yang, Y.; Ye, D.; Dai,
Z.; Liu, Y.; Ju, H. Photo-tearable tape close-wrapped upconversion
nanocapsules for near-infrared modulated efficient siRNA delivery
and therapy. Biomaterials 2018, 163, 55−66.
(48) Wang, S. J.; Tian, Y.; Tian, W.; Sun, J.; Zhao, S.; Liu, Y.; Wang,
C. Y.; Tang, Y. X.; Ma, X. Q.; Teng, Z. G.; Lu, G. M. Selectively
Sensitizing Malignant Cells to Photothermal Therapy Using a CD44-
Targeting Heat Shock Protein 72 Depletion Nanosystem. ACS Nano
2016, 10 (9), 8578−8590.
(49) Chu, C.; Ren, E.; Zhang, Y.; Yu, J.; Lin, H.; Pang, X.; Zhang, Y.;
Liu, H.; Qin, Z.; Cheng, Y.; Wang, X.; Li, W.; Kong, X.; Chen, X.; Liu,
G. Zinc(II)-Dipicolylamine Coordination Nanotheranostics: Toward
Synergistic Nanomedicine by Combined Photo/Gene Therapy.
Angew. Chem., Int. Ed. Engl. 2019, 58 (1), 269−272.
(50) Taghavi, S.; Abnous, K.; Taghdisi, S. M.; Ramezani, M.;
Alibolandi, M. Hybrid carbon-based materials for gene delivery in
cancer therapy. J. Controlled Release 2020, 318, 158−175.
(51) Zhang, Y.; Yang, L.; Yang, C.; Liu, J. Recent advances of smart
acid-responsive gold nanoparticles in tumor therapy. Wiley Interdiscip
Rev. Nanomed Nanobiotechnol 2020, 12, No. e1619.
(52) Wang, Z. Q.; Wang, L. C.; Prabhakar, N.; Xing, Y. X.;
Rosenholm, J. M.; Zhang, J. X.; Cai, K. Y. CaP coated mesoporous
polydopamine nanoparticles with responsive membrane permeation
ability for combined photothermal and siRNA therapy. Acta
Biomaterialia 2019, 86, 416−428.
(53) Altieri, D. C. Validating survivin as a cancer therapeutic target.
Nat. Rev. Cancer 2003, 3 (1), 46−54.
(54) Chu, C.; Su, M.; Zhu, J.; Li, D.; Cheng, H.; Chen, X.; Liu, G.
Metal-Organic Framework Nanoparticle-Based Biomineralization: A
New Strategy toward Cancer Treatment. Theranostics 2019, 9 (11),
3134−3149.
(55) Shao, J.; Xie, H.; Huang, H.; Li, Z.; Sun, Z.; Xu, Y.; Xiao, Q.;
Yu, X. F.; Zhao, Y.; Zhang, H.; Wang, H.; Chu, P. K. Biodegradable
black phosphorus-based nanospheres for in vivo photothermal cancer
therapy. Nat. Commun. 2016, 7, 12967.
(56) Wang, H.; Yang, X. Z.; Shao, W.; Chen, S. C.; Xie, J. F.; Zhang,
X. D.; Wang, J.; Xie, Y. Ultrathin Black Phosphorus Nanosheets for
ACS Mater. Au XXXX, XXX, XXX−XXX
ACS Materials Au pubs.acs.org/materialsau
Efficient Singlet Oxygen Generation. J. Am. Chem. Soc. 2015, 137
(35), 11376−11382.
(57) Wang, H.; Zhong, L.; Liu, Y.; Xu, X.; Xing, C.; Wang, M.; Bai,
S. M.; Lu, C. H.; Yang, H. H. A black phosphorus nanosheet-based
siRNA delivery system for synergistic photothermal and gene therapy.
Chem. Commun. (Camb) 2018, 54 (25), 3142−3145.
(58) Wang, Z.; Li, S.; Zhang, M.; Ma, Y.; Liu, Y.; Gao, W.; Zhang, J.;
Gu, Y. Laser-Triggered Small Interfering RNA Releasing Gold
Nanoshells against Heat Shock Protein for Sensitized Photothermal
Therapy. Adv. Sci. (Weinh) 2017, 4 (2), 1600327.
(59) Tang, W.; Han, L.; Lu, X.; Wang, Z.; Liu, F.; Li, Y.; Liu, S.; Liu,
S.; Tian, R.; Liu, J.; Ding, B. A Nucleic Acid/Gold Nanorod-Based
Nanoplatform for Targeted Gene Editing and Combined Tumor
Therapy. ACS Appl. Mater. Interfaces 2021, 13 (18), 20974−20981.
(60) Yang, Y.; Han, Y.; Sun, Q.; Cheng, J.; Yue, C.; Liu, Y.; Song, J.;
Jin, W.; Ding, X.; de la Fuente, J. M.; Ni, J.; Wang, X.; Cui, D. Au-
siRNA@ aptamer nanocages as a high-efficiency drug and gene
delivery system for targeted lung cancer therapy. J. Nanobiotechnol.
2021, 19 (1), 54.
(61) Deng, S. H.; Wang, S. Y.; Xiao, Z. C.; Cheng, D.
Unprotonatable and ROS-Sensitive Nanocarrier for NIR Spatially
Activated siRNA Therapy with Synergistic Drug Effect. SMALL 2022,
18 (41), 2203823.
(62) Li, G. C.; Mivechi, N. F.; Weitzel, G. Heat-Shock Proteins,
Thermotolerance, And Their Relevance to Clinical Hyperthermia. Int.
J. Hyperthermia 1995, 11 (4), 459−488.
(63) Ding, F.; Gao, X. H.; Huang, X. G.; Ge, H.; Xie, M.; Qian, J.
W.; Song, J.; Li, Y. H.; Zhu, X. Y.; Zhang, C. Polydopamine-coated
nucleic acid nanogel for siRNA-mediated low-temperature photo-
thermal therapy. Biomaterials 2020, 245, 119976.
(64) Dang, J.; Ye, H.; Li, Y.; Liang, Q.; Li, X.; Yin, L. Multivalency-
assisted membrane-penetrating siRNA delivery sensitizes photo-
thermal ablation via inhibition of tumor glycolysis metabolism.
Biomaterials 2019, 223, 119463.
(65) Chen, W. H.; Luo, G. F.; Lei, Q.; Hong, S.; Qiu, W. X.; Liu, L.
H.; Cheng, S. X.; Zhang, X. Z. Overcoming the Heat Endurance of
Tumor Cells by Interfering with the Anaerobic Glycolysis Metabolism
for Improved Photothermal Therapy. ACS Nano 2017, 11 (2), 1419−
1431.
(66) Kogel, D.; Linder, B.; Brunschweiger, A.; Chines, S.; Behl, C. At
the Crossroads of Apoptosis and Autophagy: Multiple Roles of the
Co-Chaperone BAG3 in Stress and Therapy Resistance of Cancer.
Cells 2020, 9 (3), 574.
(67) Rauch, J. N.; Tse, E.; Freilich, R.; Mok, S. A.; Makley, L. N.;
Southworth, D. R.; Gestwicki, J. E. BAG3 Is a Modular, Scaffolding
Protein that physically Links Heat Shock Protein 70 (Hsp70) to the
Small Heat Shock Proteins. J. Mol. Biol. 2017, 429 (1), 128−141.
(68) Fang, X.; Bogomolovas, J.; Wu, T. B.; Zhang, W.; Liu, C. Z.;
Veevers, J.; Stroud, M. J.; Zhang, Z. Y.; Ma, X. L.; Mu, Y. X.; Lao, D.
H.; Dalton, N. D.; Gu, Y. S.; Wang, C. L.; Wang, M.; Liang, Y.; Lange,
S.; Ouyang, K. F.; Peterson, K. L.; Evans, S. M.; Chen, J. Loss-of-
function mutations in co-chaperone BAG3 destabilize small HSPs and
cause cardiomyopathy. J. Clin. Invest. 2017, 127 (8), 3189−3200.
(69) Wang, B. K.; Yu, X. F.; Wang, J. H.; Li, Z. B.; Li, P. H.; Wang,
H. Y.; Song, L.; Chu, P. K.; Li, C. Z. Gold-nanorods-siRNA nanoplex
for improved photothermal therapy by gene silencing. Biomaterials
2016, 78, 27−39.
(70) He, X.; Hao, Y.; Chu, B.; Yang, Y.; Sun, A.; Shi, K.; Yang, C.;
Zhou, K.; Qu, Y.; Li, H.; Qian, Z. Redox–activatable photothermal
therapy and enzyme-mediated tumor starvation for synergistic cancer
therapy. Nano Today 2021, 39, 101174.
(71) Israelsen, W. J.; Vander Heiden, M. G. Pyruvate kinase:
Function, regulation and role in cancer. Semin. Cell Dev. Biol. 2015,
43, 43−51.
(72) Melamed, J. R.; Edelstein, R. S.; Day, E. S. Elucidating the
Fundamental Mechanisms of Cell Death Triggered by Photothermal
Therapy. ACS Nano 2015, 9 (1), 6−11.
O https://doi.org/10.1021/acsmaterialsau.3c00035
Review
(73) Martin, S. J.; Henry, C. M.; Cullen, S. P. A Perspective on
Mammalian Caspases as Positive and Negative Regulators of
Inflammation. Mol. Cell 2012, 46 (4), 387−397.
(74) Bonfil, R. D.; Bustuoabad, O. D.; Ruggiero, R. A.; Meiss, R. P.;
Pasqualini, C. D. Tumor Necrosis Can Facilitate the Appearance of
Metastases. Clin. Exp. Metastasis 1988, 6 (2), 121−129.
(75) Li, W. T.; Peng, J. R.; Tan, L. W.; Wu, J.; Shi, K.; Qu, Y.; Wei,
X. W.; Qian, Z. Y. Mild photothermal therapy/photodynamic
therapy/chemotherapy of breast cancer by Lyp-1 modified Docetax-
el/IR820 Co-loaded micelles. Biomaterials 2016, 106, 119−133.
(76) Ding, Y.; Du, C.; Qian, J. W.; Dong, C. M. NIR-Responsive
Polypeptide Nanocomposite Generates NO Gas, Mild Photothermia,
and Chemotherapy to Reverse Multidrug-Resistant Cancer. Nano
Lett. 2019, 19 (7), 4362−4370.
(77) Yang, Y.; Zhu, W.; Dong, Z.; Chao, Y.; Xu, L.; Chen, M.; Liu,
Z. 1D Coordination Polymer Nanofibers for Low-Temperature
Photothermal Therapy. Adv. Mater. 2017, 29 (40), 1703588.
(78) Zhou, J.; Li, M. H.; Hou, Y. H.; Luo, Z.; Chen, Q. F.; Cao, H.
X.; Huo, R. L.; Xue, C. C.; Sutrisno, L.; Hao, L.; Cao, Y.; Ran, H. T.;
Lu, L.; Li, K.; Cai, K. Y. Engineering of a Nanosized Biocatalyst for
Combined Tumor Starvation and Low-Temperature Photothermal
Therapy. ACS Nano 2018, 12 (3), 2858−2872.
(79) Ding, F.; Mou, Q. B.; Ma, Y.; Pan, G. F.; Guo, Y. Y.; Tong, G.
S.; Choi, C. H. J.; Zhu, X. Y.; Zhang, C. A Crosslinked Nucleic Acid
Nanogel for Effective siRNA Delivery and Antitumor Therapy. Angew.
Chem.-Int. Ed. 2018, 57 (12), 3064−3068.
(80) Jang, Y.; Kim, D.; Lee, H.; Jang, H.; Park, S.; Kim, G. E.; Lee,
H. J.; Kim, H. J.; Kim, H. Development of an ultrasound triggered
nanomedicine-microbubble complex for chemo-photodynamic-gene
therapy. Nanomedicine: Nanotechnology, Biology and Medicine 2020,
27, 102194.
(81) Pu, Y.; Wu, W.; Xiang, H.; Chen, Y.; Xu, H. CRISPR/Cas9-
based genome editing for multimodal synergistic cancer nanotherapy.
Nano Today 2023, 48, 101734.
(82) Haque, F.; Shu, D.; Shu, Y.; Shlyakhtenko, L. S.; Rychahou, P.
G.; Mark Evers, B.; Guo, P. Ultrastable synergistic tetravalent RNA
nanoparticles for targeting to cancers. Nano Today 2012, 7 (4), 245−
257.
(83) Jin, Y.; Wang, H.; Li, X.; Zhu, H.; Sun, D.; Sun, X.; Liu, H.;
Zhang, Z.; Cao, L.; Gao, C.; Wang, H.; Liang, X.-J.; Zhang, J.; Yang,
X. Multifunctional DNA Polymer-Assisted Upconversion Therapeutic
Nanoplatform for Enhanced Photodynamic Therapy. ACS Appl.
Mater. Interfaces 2020, 12 (24), 26832−26841.
(84) Lal, S.; Clare, S. E.; Halas, N. J. Nanoshell-Enabled
Photothermal Cancer Therapy: Impending Clinical Impact. Acc.
Chem. Res. 2008, 41 (12), 1842−1851.
(85) Liu, T.; Li, W.; Lu, W.; Chen, M.; Luo, M.; Zhang, C.; Li, Y.;
Qin, G.; Shi, D.; Xiao, B.; Qiu, H.; Yu, W.; Kang, L.; Kang, T.; Huang,
W.; Yu, X.; Wu, X.; Deng, W. RBFOX3 Promotes Tumor Growth and
Progression via hTERT Signaling and Predicts a Poor Prognosis in
Hepatocellular Carcinoma. Theranostics 2017, 7 (12), 3138−3154.
(86) Chen, L.; Chen, C.; Chen, W.; Li, K.; Chen, X.; Tang, X.; Xie,
G.; Luo, X.; Wang, X.; Liang, H.; Yu, S. Biodegradable Black
Phosphorus Nanosheets Mediate Specific Delivery of hTERT siRNA
for Synergistic Cancer Therapy. ACS Appl. Mater. Interfaces 2018, 10
(25), 21137−21148.
(87) Qiao, S.; Xin, F.; Wu, M.; Zheng, Y.; Zhao, B.; Zhang, C.; Liu,
X.; Wei, Z.; Liu, J. A remotely controlled NIR-II photothermal-
sensitive transgene system for hepatocellular carcinoma synergistic
therapy. J. Mater. Chem. B 2021, 9 (25), 5083−5091.
(88) Zhou, R.; Liu, X.; Wu, Y.; Xiang, H.; Cao, J.; Li, Y.; Yin, W.; Zu,
Y.; Li, J.; Liu, R.; Zhao, F.; Liu, Z.; Chen, C.; Gu, Z.; Yan, L.; Zhao, Y.
Suppressing the Radiation-Induced Corrosion of Bismuth Nano-
particles for Enhanced Synergistic Cancer Radiophototherapy. ACS
Nano 2020, 14 (10), 13016−13029.
(89) Wang, J.; He, H.; Xu, X.; Wang, X.; Chen, Y.; Yin, L. Far-red
light-mediated programmable anti-cancer gene delivery in coopera-
tion with photodynamic therapy. Biomaterials 2018, 171, 72−82.
ACS Mater. Au XXXX, XXX, XXX−XXX
ACS Materials Au pubs.acs.org/materialsau
(90) Zhang, X.; Yang, Y.; Kang, T.; Wang, J.; Yang, G.; Yang, Y.; Lin,
X.; Wang, L.; Li, K.; Liu, J.; Ni, J.-S. NIR-II Absorbing Semi-
conducting Polymer-Triggered Gene-Directed Enzyme Prodrug
Therapy for Cancer Treatment. Small 2021, 17 (23), 2100501.
(91) Revia, R. A.; Zhang, M. Magnetite nanoparticles for cancer
diagnosis, treatment, and treatment monitoring: recent advances.
Mater. Today (Kidlington) 2016, 19 (3), 157−168.
(92) Tay, Z. W.; Chandrasekharan, P.; Chiu-Lam, A.; Hensley, D.
W.; Dhavalikar, R.; Zhou, X. Y.; Yu, E. Y.; Goodwill, P. W.; Zheng, B.;
Rinaldi, C.; Conolly, S. M. Magnetic Particle Imaging-Guided Heating
in Vivo Using Gradient Fields for Arbitrary Localization of Magnetic
Hyperthermia Therapy. ACS Nano 2018, 12 (4), 3699−3713.
(93) Kumar, C. S.; Mohammad, F. Magnetic nanomaterials for
hyperthermia-based therapy and controlled drug delivery. Adv. Drug
Deliv Rev. 2011, 63 (9), 789−808.
(94) Liu, X.; Zhang, Y.; Wang, Y.; Zhu, W.; Li, G.; Ma, X.; Zhang,
Y.; Chen, S.; Tiwari, S.; Shi, K.; Zhang, S.; Fan, H. M.; Zhao, Y. X.;
Liang, X. J. Comprehensive understanding of magnetic hyperthermia
for improving antitumor therapeutic efficacy. Theranostics 2020, 10
(8), 3793−3815.
(95) Jordan, A.; Scholz, R.; Wust, P.; Fahling, H.; Felix, R. Magnetic
fluid hyperthermia (MFH): Cancer treatment with AC magnetic field
induced excitation of biocompatible superparamagnetic nanoparticles.
J. Magn. Magn. Mater. 1999, 201, 413−419.
(96) Hergt, R.; Dutz, S. Magnetic particle hyperthermia�
biophysical limitations of a visionary tumour therapy. J. Magn.
Magn. Mater. 2007, 311 (1), 187−192.
(97) Cheng, L.; Ke, Y.; Yu, S.; Jing, J. Co-delivery of doxorubicin and
recombinant plasmid pHSP70-Plk1-shRNA by bacterial magneto-
somes for osteosarcoma therapy. Int. J. Nanomedicine 2016, 11, 5277−
5286.
(98) Ito, A.; Shinkai, M.; Honda, H.; Kobayashi, T. Heat-inducible
TNF-alpha gene therapy combined with hyperthermia using magnetic
nanoparticles as a novel tumor-targeted therapy. Cancer Gene Ther.
2001, 8 (9), 649−654.
(99) Walther, W.; Stein, U. Heat-responsive gene expression for
gene therapy. Adv. Drug Deliv Rev. 2009, 61 (7−8), 641−9.
(100) Zhang, Z. Q.; Song, S. C. Multiple hyperthermia-mediated
release of TRAIL/SPION nanocomplex from thermosensitive
polymeric hydrogels for combination cancer therapy. Biomaterials
2017, 132, 16−27.
(101) Moros, M.; Idiago-Lopez, J.; Asin, L.; Moreno-Antolin, E.;
Beola, L.; Grazu, V.; Fratila, R. M.; Gutierrez, L.; de la Fuente, J. M.
Triggering antitumoural drug release and gene expression by magnetic
hyperthermia. Adv. Drug Deliv Rev. 2019, 138, 326−343.
(102) Yu, S.; Yi, M.; Qin, S.; Wu, K. Next generation chimeric
antigen receptor T cells: safety strategies to overcome toxicity. Mol.
Cancer 2019, 18 (1), 125.
(103) Yuan, C.; An, Y.; Zhang, J.; Li, H.; Zhang, H.; Wang, L.;
Zhang, D. Magnetic nanoparticles for targeted therapeutic gene
delivery and magnetic-inducing heating on hepatoma. Nanotechnology
2014, 25 (34), 345101.
(104) Wang, Z.; Chang, Z. M.; Lu, M. M.; Shao, D.; Yue, J.; Yang, D.
A.; Zheng, X.; Li, M. Q.; He, K.; Zhang, M.; Chen, L.; Dong, W. F.
Shape-controlled magnetic mesoporous silica nanoparticles for
magnetically-mediated suicide gene therapy of hepatocellular
carcinoma. Biomaterials 2018, 154, 147−157.
(105) Noh, S.-h.; Moon, S. H.; Shin, T.-H.; Lim, Y.; Cheon, J.
Recent advances of magneto-thermal capabilities of nanoparticles:
From design principles to biomedical applications. Nano Today 2017,
13, 61−76.
(106) Court, K. A.; Hatakeyama, H.; Wu, S. Y.; Lingegowda, M. S.;
Rodriguez-Aguayo, C.; Lopez-Berestein, G.; Ju-Seog, L.; Rinaldi, C.;
Juan, E. J.; Sood, A. K.; Torres-Lugo, M. HSP70 Inhibition
Synergistically Enhances the Effects of Magnetic Fluid Hyperthermia
in Ovarian Cancer. Mol. Cancer Ther 2017, 16 (5), 966−976.
(107) Bader, A. G.; Brown, D.; Winkler, M. The Promise of
MicroRNA Replacement Therapy. Cancer Res. 2010, 70 (18), 7027−
7030.
P https://doi.org/10.1021/acsmaterialsau.3c00035
Review
(108) Roush, S.; Slack, F. J. The let-7 family of microRNAs. Trends
Cell Biol. 2008, 18 (10), 505−16.
(109) Yin, P. T.; Shah, B. P.; Lee, K. B. Combined magnetic
nanoparticle-based microRNA and hyperthermia therapy to enhance
apoptosis in brain cancer cells. Small 2014, 10 (20), 4106−4112.
(110) Yin, P. T.; Shah, S.; Pasquale, N. J.; Garbuzenko, O. B.;
Minko, T.; Lee, K.-B. Stem cell-based gene therapy activated using
magnetic hyperthermia to enhance the treatment of cancer.
Biomaterials 2016, 81, 46−57.
(111) Kennedy, L. B.; Salama, A. K. S. A review of cancer
immunotherapy toxicity. CA Cancer J. Clin 2020, 70 (2), 86−104.
(112) Bhatia, A.; Kumar, Y. Cellular and molecular mechanisms in
cancer immune escape: a comprehensive review. Expert Rev. Clin
Immunol 2014, 10 (1), 41−62.
(113) Dai, H.; Fan, Q.; Wang, C. Recent applications of
immunomodulatory biomaterials for disease immunotherapy. Explo-
ration 2022, 2 (6), 20210157−20210157.
(114) Jiang, H.; Rivera-Molina, Y.; Gomez-Manzano, C.; Clise-
Dwyer, K.; Bover, L.; Vence, L. M.; Yuan, Y.; Lang, F. F.; Toniatti, C.;
Hossain, M. B.; Fueyo, J. Oncolytic Adenovirus and Tumor-Targeting
Immune Modulatory Therapy Improve Autologous Cancer Vacci-
nation. Cancer Res. 2017, 77 (14), 3894−3907.
(115) Lang, F. F.; Conrad, C.; Gomez-Manzano, C.; Yung, W. K. A.;
Sawaya, R.; Weinberg, J. S.; Prabhu, S. S.; Rao, G.; Fuller, G. N.;
Aldape, K. D.; Gumin, J.; Vence, L. M.; Wistuba, I.; Rodriguez-
Canales, J.; Villalobos, P. A.; Dirven, C. M. F.; Tejada, S.; Valle, R. D.;
Alonso, M. M.; Ewald, B.; Peterkin, J. J.; Tufaro, F.; Fueyo, J. Phase I
Study of DNX-2401 (Delta-24-RGD) Oncolytic Adenovirus:
Replication and Immunotherapeutic Effects in Recurrent Malignant
Glioma. J. Clin Oncol 2018, 36 (14), 1419−1427.
(116) Sheridan, C. First oncolytic virus edges towards approval in
surprise vote. Nat. Biotechnol. 2015, 33 (6), 569−570.
(117) Heiber, J.; Hyun, J.; Obuchi, M.; Barber, G. N. Development
of recombinant vesicular stomatitis virus for use as an oncolytic vector
in cancer therapy. Cytokine 2009, 48 (1−2), 47−47.
(118) Yin, Y.; Li, X.; Ma, H.; Zhang, J.; Yu, D.; Zhao, R.; Yu, S.; Nie,
G.; Wang, H. In Situ Transforming RNA Nanovaccines from
Polyethylenimine Functionalized Graphene Oxide Hydrogel for
Durable Cancer Immunotherapy. Nano Lett. 2021, 21 (5), 2224−
2231.
(119) Sayour, E. J.; Grippin, A.; De Leon, G.; Stover, B.; Rahman,
M.; Karachi, A.; Wummer, B.; Moore, G.; Castillo-Caro, P.;
Fredenburg, K.; Sarkisian, M. R.; Huang, J.; Deleyrolle, L. P.;
Sahay, B.; Carrera-Justiz, S.; Mendez-Gomez, H. R.; Mitchell, D. A.
Personalized Tumor RNA Loaded Lipid-Nanoparticles Prime the
Systemic and Intratumoral Milieu for Response to Cancer
Immunotherapy. Nano Lett. 2018, 18 (10), 6195−6206.
(120) Chen, Y.; Zhang, Y.; Wang, B.; Fan, Q.; Yang, Q.; Xu, J.; Dai,
H.; Xu, F.; Wang, C. Blood Clot Scaffold Loaded with Liposome
Vaccine and siRNAs Targeting PD-L1 and TIM-3 for Effective DC
Activation and Cancer Immunotherapy. ACS Nano 2023, 17 (1),
760−774.
(121) Yu, T.; Nie, W.; Hong, Z.; He, Y.; Chen, J.; Mi, X.; Yang, S.;
Li, X.; Wang, B.; Lin, Y.; Gao, X. Synergy of Immunostimulatory
Genetherapy with Immune Checkpoint Blockade Motivates Immune
Response to Eliminate Cancer. Adv. Funct. Mater. 2021, 31 (22),
2100715.
(122) Huang, K.-W.; Hsu, F.-F.; Qiu, J. T.; Chern, G.-J.; Lee, Y.-A.;
Chang, C.-C.; Huang, Y.-T.; Sung, Y.-C.; Chiang, C.-C.; Huang, R.-L.;
Lin, C.-C.; Dinh, T. K.; Huang, H.-C.; Shih, Y.-C.; Alson, D.; Lin, C.-
Y.; Lin, Y.-C.; Chang, P.-C.; Lin, S.-Y.; Chen, Y. Highly efficient and
tumor-selective nanoparticles for dual-targeted immunogene therapy
against cancer. Science Advances 2020, 6 (3), No. eaax5032.
(123) Li, Y.; Zhao, R.; Cheng, K.; Zhang, K.; Wang, Y.; Zhang, Y.;
Li, Y.; Liu, G.; Xu, J.; Xu, J.; Anderson, G. J.; Shi, J.; Ren, L.; Zhao, X.;
Nie, G. Bacterial Outer Membrane Vesicles Presenting Programmed
Death 1 for Improved Cancer Immunotherapy via Immune Activation
and Checkpoint Inhibition. ACS Nano 2020, 14 (12), 16698−16711.
ACS Mater. Au XXXX, XXX, XXX−XXX