12.12.

2023

Stem Cell Therapeutics for

Cancer

Video 1-Mechanism of Action of Cell-based Therapies

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Hematopoietic Stem Cells in Treatment of Cancer

Hematopoietic stem cell transplantation

• It is the process of transplanting multipotent

hematopoietic stem cells, mostly from peripheral blood,
bone marrow or umbilical cord blood. Hematopoietic
stem cell transplantation was first used in the treatment
of some types of cancer, but today it is widely used for
the treatment of various autoimmune diseases.

• United States Food and Drug Administration (FDA)

has approved hematopoietic stem cell transplantation
(HSCT)

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What is Stem Cell Transplantation Used in

Treatment?

• Stem cell transplantation has been approved for the

treatment of various cancers: multiple myeloma,
leukemia and some lymphomas.

• In addition, HSCT is used to treat autoimmune diseases

in many specialist clinics around the world. Perhaps the
most notable of these is multiple sclerosis. An
extremely high success rate for this treatment has been
claimed but is still considered experimental.

Cancer Consists of Three Main Stages

➢ development,
➢ growth,
➢ metastasis.

The most important factors in the initiation of cancer

are
• the epigenetic changes,
• genetic mutations in proto-oncogenes, tumor
suppressor genes, pro-apoptotic, anti-apoptotic, and
cell cycle controlling genes.

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tumor microenvironment
• growth, chemotherapy resistance, immune
escape, and tumor metastasis

• Angiogenesis --- (vascular endothelial

growth factor (VEGF and its receptors) in
tumor site is necessary for tumor growth and
metastasis
*** resistance to chemotherapies and
vasculogenic mimicry***

Advantages of MSCs For Medical

Applications in The Case of Cancer Therapy

• Some of the superiorities of using MSCs as therapeutic gene

micro-carriers:
✓ easy cell-extraction procedures
✓ their abundant proliferation capacity in vitro without
losing their main biological properties
✓ tumor-tropism,
✓ non-immunogenicity,
✓ stimulatory effect on the anti-inflammatory molecules,
✓ inhibitory effect on inflammatory responses,
✓ non-toxicity against the normal tissues, and easy
processes for the clinical use

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Approaches For Gene Therapy of Cancer

1. Engineered chimeric antigen receptor (CAR) T

cells;
CARs are engineered receptors with high specificity in
identifying tumor-related antigens.

2. Tumor vaccine (DNA vaccine),

DNA vaccines can establish anticancer immunity through the
induction of expression of a specific gene.

1- Engineered Chimeric antigen receptor T

cells (also known as CAR T cells)

• CART cells are T cells that have been genetically

engineered to produce an artificial T-cell receptor for
use in immunotherapy.

• CARs are receptor proteins that have been engineered

to give T cells the new ability to target a
specific protein.

• The receptors are chimeric because they combine both

antigen-binding and T-cell activating functions into a
single receptor.
Video 2-CAR T-Cell Therapy- How Does It Work

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CART cells for cancer therapy

• CAR-T cell therapy uses T cells engineered with CARs for cancer
therapy.

• The premise of CAR-T immunotherapy is to modify T cells to

recognize cancer cells in order to more effectively target and
destroy them.

• CAR-T cells can be either derived from T cells in a patient's own

blood (autologous) or derived from the T cells of another healthy
donor (allogeneic).

• For safety, CAR-T cells are engineered to be specific to an antigen

expressed on a tumor that is not expressed on healthy cells.

Ideal CAR Target…

• Tumor specific
• Universally expressed on only tumor cells
• Cell surface molecule
• CD19 antigen,
– Found on B cell malignant cells (Non-hodgkin lymphoma
(NHL), Acute lymphocytic leukemia (ALL), Chronic
Lymphocytic leukemia (CLL), etc)
– Expressed on early B cells but NOT stem cells

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2. Tumor vaccine (DNA vaccine)

• DNA vaccines for cancer immunotherapy are

designed to deliver one or several genes
encoding tumor antigens, thereby eliciting or
augmenting antigen-specific immune responses
against antigens that play a central role
in tumor initiation, progression and metastasis.

Cancer Vaccines
Active and specific stimulation
of immune system against cancer

Therapeutic
Increased tumor antigen recognition
10% of Reduced immune tolerance
Preventive
cases
Human Papilloma Virus (HPV) Cervical cancer
Tumor Antigens
Hepatitis B virus (HBV) Hepatocellular carcinoma
25% of
Nonpathogenic virus
cases

Vaccines
70% Tumor
Anti HPV reduced risk Associated
Anti HBV Lymphocytes
Cancer cells

Antigen presenting cells

Vaccines:
Melanoma
Prostate Cancer
Follicular lymphoma

Tumor associated lymphocytes: A type of immune cell that has moved from the blood into a tumor

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Preventive Cancer Vaccines

• Viral infections are responsible for the development of several

cancers and preventive vaccines play an important role in
reducing risk. For instance, cervical cancer and head and neck
cancer can be caused by human papilloma virus, or HPV,
whereas liver cancer can be caused by hepatitis B virus or
HBV. Several vaccines have been developed that can prevent
HBV and HPV infection and, as a result, protect against the
formation of HBV- and HPV-related cancers.
• Four of these preventive cancer vaccines have been approved by
the U.S. Food and Drug Administration (FDA).

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Preventive Cancer Vaccines

• Cervarix®: a vaccine approved for use in preventing infection by the two

strains of HPV that cause most cervical cancers, HPV types 16 and 18; can
help prevent the development of HPV-related anal, cervical, head and neck,
penile, vulvar, and vaginal cancers
• Gardasil®: a vaccine that protects against infection by HPV types 16, 18, 6,
and 11; can help prevent the development of HPV-related anal, cervical, head
and neck, penile, vulvar, and vaginal cancers
• Gardasil-9®: a vaccine approved for the prevention of infection by HPV
types 16, 18, 31, 33, 45, 52, and 58, and for the prevention of genital warts
caused by HPV types 6 or 11; can help prevent the development of HPV-
related anal, cervical, head and neck, penile, throat, vulvar, and vaginal
cancers
• Hepatitis B (HBV) vaccine (HEPLISAV-B®): a preventive vaccine that
protects against infection by the hepatitis B virus; can help prevent the
development of HBV-related liver cancer

Therapeutic Cancer Vaccines

• Each individual’s tumor is in some sense unique and has its own
distinguishing antigens. As a result, more sophisticated cancer vaccine
approaches are necessary.
• Fortunately, scientists now identify targets on patients’ tumors that can
help distinguish cancer cells from their normal cells. Sometimes these
targets are normal proteins that are produced at abnormally high levels by
cancer cells, such as prostatic acid phosphatase (PAP), which is often
overexpressed by prostate cancer cells. Taking advantage of that insight,
the sipuleucel-T vaccine was developed and received FDA approval in
2010 for the treatment of patients with advanced prostate cancer.
Additionally, virus-derived proteins expressed by virus-infected
cancer cells offer another promising source of markers that can be
targeted through vaccines.
• Another exception is Bacillus Calmette-Guérin, or BCG, a tuberculosis
vaccine that acts as a general immune stimulant. In 1990, BCG became
the first immunotherapy of any type to be approved by the FDA and
is still used for the treatment of early-stage bladder cancer.

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Therapeutic Cancer Vaccines

• Bacillus Calmette-Guérin (BCG): a vaccine that uses

weakened bacteria to stimulate the immune system;
approved for patients with early-stage bladder cancer
• Sipuleucel-T (Provenge®): a vaccine composed of
patients’ own stimulated dendritic cells; approved for
prostate cancer
Prostate Cancer vaccine: https://www.cast-pharma.com/portfolio/prostate-cancer-vaccine/

Vaccine Induced Immune Responses

1 2

3 4

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Mechanisms of Action of DNA Vaccines

Approaches For Gene Therapy of Cancer

3. Replacing the normal hematopoietic stem cells;

the normal cells are transfected with specific
chemotherapy-resistant genes then transplanted to the
patient by bone marrow transplantation.

4. Clustered regularly interspaced short

palindromic repeats (CRISPR)–Cas9 technology;

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Approaches For Gene Therapy of Cancer

5. Therapeutic genes/agents delivery;

viral and non-viral vectors, tumor-tropic cells, and

other micro-carriers are used for the transfer or
expression of therapeutic genes, agents, or even
oncolytic viruses
Delivered anticancer factors:
tumor suppressor genes, apoptosis-inducing genes,
suicide genes, regulatory agents [e.g., RNA
interference (RNAi), miRNAs], oncolytic viruses and
immunological factors (e.g., cytokines)

Therapeutic genes can be transferred as the naked

DNA or by using viral/non-viral vectors.
But the main disadvantage of this classical method
of gene delivery is its generally non-selective
nature.

Transgene MSCs selectively migrate toward the

injured and tumor site(s)

tropism to tumor sites and immunomodulatory

properties of MSCs

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The Strategies Applied for the Anti-cancer Genes/Agents

Delivery are Based on the Following Principles

1. Augmentation gene therapy

(a) expressing a gene to prompt apoptosis (e.g.,
TRAIL, mda-7, Caspases and selective short
interfering RNA (siRNA)/microRNA (miRNA)-
mediated blocking of anti- apoptotic genes),
(b) improving tumor sensitivity to chemo/ radiation
therapy,
(c) introducing a tumor suppressor gene (e.g., P53, Rb,
p16INK/CDKN2, and PTEN).

2. Gene silencing therapy:

inhibition of expression of an oncogene (C-MYC and K-
Ras) by employing an antisense (RNA/DNA).

3. Suicide gene therapy:

delivery of a converting enzyme to the site of tumor that
convert non-toxic prodrug to the toxic drug.

4. Immuno-gene therapy: increasing the

immunogenicity of the tumor cells/tissue to stimulate
immune cell response against tumor.
Video 3-Oncolytic Virus Therapy- Dynamite for Cancer Cells

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The Strategies Of Anticancer Gene Therapy Using Mesenchymal

Stromal/Stem Cells (Mscs) As Gene Vehicles

The Mechanisms of MSCs Homing to Tumor

Tissue

• Tumor cells resemble a chronic inflammation within

the tumor microenvironment by generating high
concentrations of inflammatory chemokines and
growth factors.

• Selective migration of MSCs to the tumor site is linked

to the high local concentrations of dozens of
chemoattractants and growth factors that are secreted
by tumor cells and inflammatory cells.

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The Mechanisms of MSCs Homing to Tumor

Tissue

• MSCs can migrate to sites of trauma, injury and

tumor
• gradient of chemo-attractants in the extracellular
matrix (ECM) and peripheral blood (Son et al.,
2006)
• and local factors, such as hypoxia, cytokine
environment and Toll-like receptors ligands, where
upon arrival these local factors promote MSCs to
express growth factors that accelerate tissue
regeneration (Rustad and Gurtner, 2012).

The Mechanism of Mesenchymal Stromal/Stem Cells (MSCs)

Migration/Homing Toward Injured/Cancer Tissues

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Increasing the efficiency of Tumor Tropism of

MSCs

• low-dose irradiation of tumor can increase the

recruitment of MSCs to the tumor site(s).

• irradiation increases the apoptosis and stimulates the

danger signals. The danger signals thereby induce the
production of inflammatory cytokines such as
PDGF, TNFα, CCR8, and CCR2 within the tumor
microen-vironment, and consequently enhance MSCs
swarming into the tumor location(s)

Genetically Engineered MSCs With

Anticancer Activity

• MSCs genetically modified with interferon β (IFN-β)

were injected into human melanoma mouse
xenotransplantation models which resulted in
decreased tumor growth and increased (2-times)
survival of mice in comparison with controls
(Studeny et al., 2002).

• One of the most promising therapeutic pro-apoptotic

cytokines is tumor necrosis factor (TNF)-related
apoptosis-inducing ligand (TRAIL), which
selectively induces apoptosis in cancer cells.

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Genetically Engineered MSCs With

Anticancer Activity
• IL-12, CX3CL1, INF-β, INF-α, INF-γ, IL-2, hepatocyte growth
factor antagonist NK4, pigment epithelium-derived factor,
TRAIL, and TNF-α have antitumor effect.

• Tumor necrosis factor (TNF)‐related apoptosis‐inducing ligand

(TRAIL) is the ligand for death receptors which are expressed
on the surface of tumor cells. TRAIL can initiate the caspase-
mediated apoptosis leading to inhibition of tumor growth

• MSCs and generally other normal cells are nearly resistant to

TRAIL-induced apoptosis due to very low expression of death
receptors.
• TRAIL-directed death induction can be of great advantages
to the designing of a selective cancer therapy

Genetically Engineered MSCs With

Anticancer Activity

Factor Application MSCS Host Vector Tumor model Result

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Delivery of oncolytic viruses with MSCs

• For instance, Du et al. (2017) used MSCs as a

vector for the delivery of oncolytic herpes simplex
virus (oHSV) [approved by Food and Drug
Administration (FDA) for melanoma treatment] in
human brain melanoma metastasis models in
immunodeficient and immunocompetent mice.
Authors noted that the introduced MSCs-oHSV
migrated to the site of tumor formation and
significantly prolonged the survival of mice.

3-Oncolytic Virus Therapy- Dynamite for Cancer Cells

MSCs Primed With Anticancer Drugs

Mesenchymal stem cells:

• relative resistance to cytostatic and cytotoxic
chemotherapeutic drugs
• migration ability
targeted delivery of therapeutic drugs directly to tumor
sites.

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MSCs Primed With Anticancer Drugs

• mouse bone marrow stromal cells can be a reservoir for doxorubicin
(DOX) which can subsequently be released not only in the form of
DOX metabolites but also in its original form.

• MSCs efficiently absorb and release paclitaxel (PTX) in an active form

(Pascucci et al., 2014), DOX, and gemcitabine (GCB), all having an
inhibitory effect on tongue squamous cell carcinoma (SCC154) cells
growth in vitro (Cocce et al., 2017b).

• Beside chemical drugs in soluble form, MSCs can absorb

nanomaterials containing chemotherapeutic agents. For instance,
MSCs primed with silica nanoparticle-encapsulated DOX promoted a
significant increase in the apoptosis of U251 glioma cells in vivo (Li et
al., 2011).

MSCs as Transporters of Therapeutic

Molecules

• Moreover, MSCs are capable of being reprogrammed

for transporting therapeutic molecules/proteins in the
same manner that they can carry the therapeutic genes.

• This helps to overcome the adverse effects associated

with the direct injection of drugs or other therapeutic
molecules.

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Gene-directed enzyme prodrug therapy

(GDEPT) by MSCs

• At the first stage, the genes encoding the pro- drug activating
enzymes are transferred to the tumor site using MSCs as the cell
vehicles.
• Subsequently, inactive and non-toxic prodrug is injected to the
body.
• Then, pro-drug is catalyzed by the enzymatic cleavage to the
activated form within the tumor environment.
• At the last stage, cytotoxic metabolites derived from injected and
catalyzed pro-drug are released to the tumor microenvironment
causing apoptosis, necrosis and death of the tumor cells

Table 2 | Gene-directed enzyme pro-drug therapy (GDEPT) of cancers using the

various types of mesenchymal stromal/stem cell (MSCs).

BM-MSCs, bone marrow-derived mesenchymal stem cells; AT-MSCs, adipose tissue mesenchymal stem cells;
i.v., intravenous; s.c., subcutaneous; dTRAIL, dodecameric human TRAIL; hASCs, human adipose-derived
stroma and stem cells; NSCs, neural stem cells, CDy::UPRT, fusion yeast cytosine deaminase::uracil
phosphoribosyltransferase gene.

Mesenchymal Stromal/Stem Cells: A New Era in the Cell-Based Targeted Gene Therapy of Cancer, 2017, Front. Immunol

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The Procedures For The Isolation, Culture, Gene Transfer, and In Vivo
Administration of The Mesenchymal Stromal/Stem Cells

MSC-based Approach For Cancer Therapy

Application of Mesenchymal Stem Cells for Therapeutic Agent Delivery in Anti-tumor Treatment, Front. Pharmacol., 20 March 2018
| https://doi.org/10.3389/fphar.2018.00259

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