The official journal of

INTERNATIONAL FEDERATION OF PIGMENT CELL SOCIETIES · SOCIETY FOR MELANOMA RESEARCH

PIGMENT CELL & MELANOMA

Research
The role of ferroptosis in melanoma

Ronan Talty | Marcus Bosenberg

DOI: 10.1111/pcmr.13009
Volume 35, Issue 1, Pages 18–25

If you wish to order reprints of this article,

please see the guidelines here

EMAIL ALERTS
Receive free email alerts and stay up-to-date on what is published
in Pigment Cell & Melanoma Research – click here

Submit your next paper to PCMR online at http://mc.manuscriptcentral.com/pcmr

Subscribe to PCMR and stay up-to-date with the only journal committed to publishing
basic research in melanoma and pigment cell biology

As a member of the IFPCS or the SMR you automatically get online access to PCMR. Sign up as
a member today at www.ifpcs.org or at www.societymelanomaresarch.org

To take out a personal subscription, please click here

More information about Pigment Cell & Melanoma Research at www.pigment.org
 | |
Received: 2 June 2021    Revised: 2 August 2021    Accepted: 12 August 2021

DOI: 10.1111/pcmr.13009

REVIEW

The role of ferroptosis in melanoma

Ronan Talty1  | Marcus Bosenberg1,2,3

1
Department of Pathology, Yale School of
Medicine, New Haven, CT, USA Abstract
2
Department of Dermatology, Yale School of Melanoma is the deadliest form of skin cancer. Although treatment with targeted
Medicine, New Haven, CT, USA
therapies and immune checkpoint inhibitors has dramatically improved survival in ad-
3
Department of Immunobiology, Yale School
of Medicine, New Haven, CT, USA
vanced melanoma, many patients do not benefit from these therapies or relapse after
an initial period of response. Thus, future outcomes in these categories of melanoma
Correspondence
Dr. Marcus Bosenberg, Department of
patients will depend on the identification of novel therapeutic targets and methods
Dermatology, Yale School of Medicine, P.O. to enhance existing targeted therapy and immunotherapy regimens. Ferroptosis is a
Box 208023, New Haven, CT 06520-­8 023,
USA.
newly identified form of iron-­dependent regulated cell death that is morphologically,
Email: Marcus.bosenberg@yale.edu biochemically, and genetically distinct from apoptosis, autophagy, pyroptosis, and

Funding information
National Cancer Institute, Grant/Award
Number: F30CA254246- ­01A1
necroptosis. Dysregulation of ferroptosis has been linked to the development of sev-
eral forms of cancer. This review examines ferroptosis in the context of melanoma. It
presents an overview of ferroptosis biology, summarizes and interprets the current
literature, and poses several outstanding questions and areas of future direction.

KEYWORDS

cell death, ferroptosis, immunotherapy, iron, melanoma, skin neoplasms

1 | I NTRO D U C TI O N processes including lung fibrosis, ischemia–­

Melanoma is the deadliest form of skin cancer, and it causes an esti-
mated 9,000 deaths in the United States annually (Siegel et al., 2019).
Until recently, once distant metastasis occurred, median survival and
three-­year overall survival dropped to 8 months and less than 10%,
respectively (Wilson et al., 2019). Now, treatment with dual BRAF
and MEK-­
t argeted therapies and α-­PD-­1 and α-­C TLA-­4 immune
checkpoint inhibitors dramatically improves survival in these cases
(Robert et al., 2019; Schachter et al., 2017). Responses to these
treatments, however, remain variable. Some patients do not benefit
at all, and others relapse after an initial period of response. Thus, fu-
ture outcomes in these categories of melanoma patients will depend
on the identification of novel therapeutic targets and methods to
enhance existing targeted and immunotherapy regimens.
Ferroptosis is a newly identified form of iron-­dependent regu-
lated cell death that is morphologically, biochemically, and geneti-
cally distinct from apoptosis, autophagy, pyroptosis, and necroptosis
(Dixon et al., 2012). It is implicated in a growing list of disease
reperfusion injury,
stroke, traumatic brain injury, Alzheimer's disease, and Huntington's
disease. In addition, ineffective ferroptosis is associated with tum-
origenesis, and dysregulation of ferroptosis has been linked to the
development of breast, cervical, colorectal, gastric, hepatocellu-
lar, lung, ovarian, prostate, and renal carcinomas, lymphoma, and
melanoma (Lu et al., 2017). Comprehensive reviews of ferroptosis
have been published recently (Friedmann Angeli et al., 2019; Jiang
et al., 2021; Stockwell & Jiang, 2020). This review examines current
data that support roles for ferroptosis in melanoma pathogenesis,
metastasis, and treatment.
2 | FE R RO P TOS I S OV E RV I E W
The first ferroptosis-­inducing compounds, eradicator of RAS-­and
ST-­
expressing cells (erastin) and Ras Selective Lethal 3 (RSL3),
were identified prior to the first description of ferroptosis itself in
high-­
throughput screens for small molecules that effectively kill

© 2021 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd

|
18     
wileyonlinelibrary.com/journal/pcmr Pigment Cell Melanoma Res. 2022;35:18–25.
TALTY and BOSENBERG |
      19

tumorigenic cells (Dolma et al., 2003; Yang & Stockwell, 2008).
During their subsequent characterization, both erastin and RSL3
were noted to increase cellular reactive oxygen species (ROS) and
elicit specific morphological responses like mitochondrial shrink-
age in the absence of any features of other forms of cell death
such as chromatin condensation, cytoplasmic swelling, or plasma
membrane rupture. Also, selective modifiers of other forms of cell
death including inhibitors of caspases, calpains and cathepsins, cy-
clophilin D, RIPK1, and autophagy had no effect on the mode of
cell death induced by these compounds (Dolma et al., 2003; Yang &
Stockwell, 2008). On the contrary, a distinct set of small molecules
consisting of deferoxamine (an iron chelator), trolox (a Vitamin E
analog), ebselen (a glutathione peroxidase mimetic), and U0126 (a
MEK inhibitor) protected cells from erastin and RSL3-­induced cell
death. Two novel small molecules, named ferrostatin-­1 and liprox-
statin-­1, also inhibited this form of cell death with extremely high
selectivity (Dixon et al., 2012; Friedmann Angeli et al., 2014).
It is now known that ferroptosis occurs through three major
steps: reduced activity of the cysteine-­
glutamate antiporter
(System Xc-­), inhibition of glutathione peroxidase 4 (GPX4), and iron-­
dependent generation of lipid peroxides. The relationship between
these key processes is illustrated in Figure 1. Briefly, System Xc-­ is a
cell surface antiporter comprised of SLC7A11 and SLC3A2 that im-
ports extracellular cystine in exchange for intracellular glutamate.
Under normal conditions, both of these amino acids are necessary
for the synthesis of reduced glutathione, which is used by GPX4 to
reduce lipid (hydro)peroxides. Inhibition of this antiporter by com-
pounds like erastin, sulfasalazine, or sorafenib leads to decreased
synthesis of reduced glutathione for use by GPX4, while com-
pounds like RSL3 and FIN56 inhibit GPX4 directly. In either case,
lipid peroxides accumulate and trigger cell death by ferroptosis.
The generation of lipid peroxides can occur through enzymatic and
nonenzymatic pathways. Enzymatic lipid peroxidation is performed
mainly by lipoxygenases (LOXs) and NADPH oxidases (NOXs) that
often depend on iron as a cofactor, whereas nonenzymatic lipid
peroxidation occurs through an autoxidation reaction that uses free
iron as a substrate for Fenton-­like chemistry (Conrad & Pratt, 2019).
Consequently, processes that increase the cellular availability of
free iron, such as ferritinophagy mediated by NCOA4, can promote
ferroptosis, and iron chelators like deferoxamine (DFO), which re-
duce free iron, inhibit ferroptosis. Ferrostatin-­1, liproxstatin-­1, and
vitamin E act as radical-­trapping agents (RTAs) to inhibit the gener-
ation of lipid peroxides. A broader list of ferroptosis inducers and
inhibitors is presented in Table 1.
Several other pathways are now recognized to promote ferro-
ptosis resistance. First, ferroptosis suppressor protein 1 (FSP1), re-
named from apoptosis-­inducing mitochondria associated 2 (AIFM2),
conferred protection against ferroptosis in both a CRISPR-­
C as9
loss of function screen with RSL3 treatment and a cDNA overex-
pression screen in GPX4-­deficient cells (Bersuker et al., 2019; Doll

F I G U R E 1   An overview of ferroptosis, which is triggered by the toxic accumulation of lipid peroxides. These are generated through
a combination of iron-­mediated Fenton chemistry and the enzymes ALOX, NOX, and LPCAT. System Xc imports cystine into the cell to
support the synthesis of reduced glutathione (GSH). GSH is used by GPX4 to reduce lipid peroxides and prevent their accumulation. The
NRF2, hippo, mevalonate synthesis, and transsulfuration pathways are secondary pathways that can supplement the action of GPX4.
Ferroptosis inducers (red) and inhibitors (green) manipulate various points of these pathways. AA—­arachidonic acid; ACSL4—­acyl-­CoA
synthetase long-­chain family member 4; ALOX—­arachidonate lipoxygenase; Cys—­cysteine; CoQ—­coenzyme Q; FPN—­ferroportin; FSP1—­
ferroptosis suppressor 1; GCL—­glutamate cysteine ligase; Glu—­glutamate; GPX4—­glutathione peroxidase 4; GS—­glutamine synthetase;
GSH—­reduced glutathione; GS-­SG—­oxidized glutathione; LPCAT—­lysophosphatidylcholine acyl transferase; NCOA4—­nuclear receptor
coactivator 4; NOX—­NADPH oxidase; NRF2—­nuclear factor erythroid-­2-­related factor 2; PROM2—­prominin 2; PUFA—­polyunsaturated fatty
acid; RSL3—­R AS selective lethal 3; SLC3A2—­solute carrier family 3 member 2; SLC7A11—­solute carrier family 7 member 11; TAZ—­t afazzin;
TF—­transferrin; YAP—­yes-­associated protein
 |
20       TALTY and BOSENBERG
Mechanism Compounds
Ferroptosis inducers
FSP1-­CoQ10 -­NAD(P)H axis iFSP1
GPX4 inhibition Altretamine, DPIs, FIN56, JKE−1674, ML162, ML210, RSL3
GSH depletion Buthionine sulfoximine (BSO)
Iron overload Lapatinib, Salinomycin, Siramesine
Iron oxidation FINO2
System Xc-­inhibition Cyst(e)inase, DPI2, Erastin, Imidazole ketone erastin, Piperazine
erastin, Sorafenib, Sulfasalazine
Ferroptosis Inhibitors
Iron chelation Ciclopirox, Deferoxamine, Deferiprone
MEK inhibition U0126
Radical trapping a-­tocopherol, Ferrostatin-­1, Liproxstatin-­1
et al., 2019). FSP1 catalyzes the regeneration of CoQ10, which then
traps lipid peroxyl radicals. As another example, the nuclear factor
erythroid-­2-­related factor 2 (NRF2) transcription factor is a master
regulator of the antioxidant response. Many proteins responsible
for regulating iron metabolism (e.g., FTH, FTL, and SLC40A1) and
preventing lipid peroxidation (e.g., SLC7A11, GPX4, and FSP1) have
been identified as NRF2 target genes (Dodson et al., 2019). Other
pathways that regulate aspects of ferroptosis include hippo, meva-
lonate synthesis, transsulfuration, and prominin2-­
exosome path-
ways (Brown et al., 2019; Hayano et al., 2016; Warner et al., 2000;
Wu et al., 2019).
3 |  M E L A N O M A D E D I FFE R E NTI ATI O N
S TAT U S CO R R E L ATE S W ITH FE R RO P TOS I S
S E N S ITI V IT Y
Cellular dedifferentiation is a hallmark of cancer initiation, progres-
sion, and metastasis, and it can also drive targeted therapy resist-
ance (Gupta et al., 2019). Melanoma cells, in particular, maintain a
highly plastic phenotype, possibly due to their neural crest origin,
a transient, stem-­cell-­like embryonic cell population that differenti-
ates into diverse tissue types ranging from neurons and glia to en-
docrine cells and cartilage (Sauka-­Spengler & Bronner-­Fraser, 2008).
In clinical practice, dedifferentiation allows melanomas to develop
resistance to both BRAF and MAPK inhibition associated with down-
regulation of MITF, the master regulator of melanocyte differentia-
tion (Hugo et al., 2015; Müller et al., 2014). Furthermore, decreased
MITF expression has been shown to enhance recruitment of immu-
nosuppressive myeloid immune cells into the tumor microenviron-
ment to facilitate immune escape and tumor growth (Riesenberg
et al., 2015).
In 2018, Tsoi et al., reasoned that dedifferentiation might con-
fer susceptibility to novel therapies despite promoting resistance
to many standard interventions. To examine this, they performed
hierarchical clustering and principal component analysis of expres-
sion profiles from 53 human melanoma cells lines, including paired
TA B L E 1   A summary of common
ferroptosis inducers and inhibitors
organized by their mechanisms
acquired resistance sub-­
lines, established from patient biopsies.
The cell lines clustered into four distinct classes according to their
differentiation status: (1) melanocytic, (2) transitory, 3) neural crest
like, and 4) undifferentiated. Their analysis of these clusters revealed
consistent, specific dedifferentiation sequences that occur in re-
sponse to BRAF inhibition, MAPK inhibition, and immunotherapy and
confer resistance. They paired their clusters with publicly available
data from the Cancer Therapeutics Response Portal (CTRP), filtered
for drugs that exhibited cluster-­
specific sensitivity, and demon-
strated that dedifferentiation status positively correlated with sensi-
tivity to the ferroptosis inducers erastin, RSL3, ML162, and ML210.
After confirming these findings in their own cell lines, they tested
the combined effect of BRAF inhibitors and ferroptosis inducers in
vitro and showed a dramatic decrease in the number of cells persist-
ing after treatment. This effect, they reasoned, likely occurs because
each agent optimally targets melanoma cells at a distinct differentia-
tion status. Finally, they showed that the proinflammatory cytokines
TNF-­α and IFN-­γ induce dedifferentiation and potentiate lethality
upon treatment with ferroptosis inducers. These data suggest that
ferroptosis inducers could potentially promote antitumor immunity
by killing dedifferentiated melanoma cells and preventing their im-
munosuppressive actions. What remains unknown, however, is how
dedifferentiation leads to increased ferroptosis sensitivity. It would
be relevant to examine in subsequent experiments whether TNF-­α
and IFN-­γ elicit downregulation of System Xc-­ or glutathione synthe-
sis enzymes or whether an unrelated mechanism alters ferroptosis
sensitivity.
4 | TH E R A PY- ­R E S I S TA NT M E L A N O M A
C E LL S ACQ U I R E A G PX4 - ­D E PE N D E NT
PE R S I S TE R S TATE
Efforts to understand mechanisms of therapy resistance by cancer
cells have widely focused on genetic mutations that confer drug
resistance such as altered binding sites, compensatory signaling
pathways, and upregulation of efflux pumps. Emerging evidence,
TALTY and BOSENBERG
however, underscores the importance of non-­mutational mecha-
nisms of resistance in cancer cells (Boumahdi & de Sauvage, 2020).
Such changes in cell plasticity can enable them to either wholly resist
initial treatment or maintain a surviving “persister” cell population
that can drive disease recurrence. Specific examples of this plasticity
include epithelial–­mesenchymal transition, neuroendocrine trans-
differentiation, and dedifferentiation-­based phenotypic discussed
previously (Tsoi et al., 2018b). Cell identity conversions, chromatin
remodeling and epigenetic transformation, microenvironment con-
tributions, and the existence of slow-­cycling cells are all proposed
mechanisms for how these plasticity changes might occur in the set-
ting of cancer treatment.
Two studies implicated the central ferroptosis gene, GPX4, as
a potential driver of cell plasticity. The first explored why high-­
mesenchymal cancer cell states confer resistance to multiple
treatment modalities across diverse cancer lineages (Viswanathan
et al., 2017). The group used publicly available gene expression
data sets to evaluate shared characteristics between multiple
high-­
m esenchymal cancer cell lines. They demonstrated that
these cell lines exhibit increased synthesis of polyunsaturated
lipids which serve as substrates for lipoxygenase enzymes, and
this change in lipid production forces a dependence on the en-
zyme GPX4 to prevent the accumulation of lipid peroxides and
ferroptosis execution. This specifically occurred in BRAF-­m utant
melanoma treated with TGFβ to upregulate AXL, downregulate
MITF, and induce targeted therapy and immunotherapy resis-
tance. Importantly, loss of GPX4 induced cell death in vitro in
these therapy-­resistant cells via ferroptosis and greatly impaired
tumor growth in vivo. The second study determined that a similar
dependence on GPX4 for survival occurs in residual populations of
cancer cells that persist after a broad range of drug treatments, in-
cluding A375 melanoma cells treated with vemurafenib (Hangauer
et al., 2017). Once again, they showed that loss of GPX4 induces
death of persister cells via ferroptosis in vitro and prevents tumor
relapse after treatment in vivo. These studies implicate the ferro-
ptosis pathway as a molecular driver of cell plasticity and treat-
ment resistance in the context of melanoma. Notably, it remains
unknown whether immune checkpoint inhibitors also produce a
residual population of GPX4-­d ependent persister cells. Such in-
formation could help inform the use of ferroptosis modulators in
combination with current immunotherapy regimens for patients
with metastatic melanoma.
5 |  M I C RO R N A S R EG U L ATE FE R RO P TOS I S
I N M E L A N O M A BY TA RG E TI N G G LU TA M ATE
M E TA B O LI S M
Long before the first description of ferroptosis, it had been noted
that neurons treated with excess glutamate reduce their cystine
import, become glutathione deficient, accumulate peroxides, and
undergo cell death (Murphy et al., 1989). Glutamate-­induced cy-
totoxicity is now suspected to occur via ferroptosis and can be
|
      21
prevented with iron chelators and other ferroptosis inhibitors.
Glutaminolysis and glutamine metabolism have also been implicated
in ferroptosis regulation in other ways. For example, the glutamate
importer (SLC1A5/SLC28A1), glutaminase (GLS), and glutamic-­
oxaloacetic transaminase-­1 (GOT1) collectively facilitate glutamine
uptake and its metabolism to glutamate and α-­ketoglutarate (α-­KG),
and inactivation of any of these genes protects cells from ferropto-
sis induction (Gao et al., 2015). Additionally, the mutant Huntington
(HTT) allele found in Huntington's disease has been linked to
glutamate-­induced cytotoxicity, glutathione deficiency, disrupted
iron metabolism, and lipid peroxidation (Johnson et al., 2012). This
link has prompted an extensive study of ferroptosis in the con-
text of Huntington's disease and other neurodegenerative condi-
tions like Alzheimer's disease and Parkinson's disease (Masaldan
et al., 2019).
MicroRNAs (miRNAs) have emerged as critical regulators of
diverse biological processes such as cell death, differentiation,
proliferation, and metabolism, and two studies also demonstrate
that miRNAs can regulate ferroptosis by targeting glutamate me-
tabolism. Luo et al., (2018) performed an unbiased screen in A375
and G-­361 melanoma cells transfected with miRNA overexpressing
constructs and treated with erastin and RSL3 to discover miRNAs
that regulate ferroptosis. Overexpression of miR-­137 inhibited lipid
peroxidation, reduced iron accumulation, and rescued cells from
ferroptosis. miR-­137 accomplishes this by suppressing expression
of the SLC1A5 glutamine transporter component, and overexpres-
sion of SLC1A5 negates the ferroptosis protective effect of miR-­137.
Importantly, miR-­137 overexpression in melanoma cells produced
faster tumor growth, larger tumor volumes, and resistance to eras-
tin in vivo. miR-­9 was also identified in this screen and similarly in-
hibited lipid peroxidation, reduced iron accumulation, and rescued
cells from ferroptosis (Zhang et al., 2018). miR-­9, however, exerts no
effect on SLC1A5. Instead, it negatively regulates GOT1 expression,
and overexpression of GOT1 negates the ferroptosis protective ef-
fect of miR-­9. These studies confirm a role for miRNAs in ferroptosis
regulation, implicate glutamine metabolism in melanoma progres-
sion, and identify two potential therapeutic targets that can regu-
late ferroptosis.
6 | S E N S ITI V IT Y TO FE R RO P TOS I S
I N FLU E N C E S TH E M E TA S TATI C P OTE NTI A L
O F M E L A N O M A C E LL S
The role of oxidative stress in suppressing melanoma metastasis
is well documented, and the lymphatic system is the route of me-
tastasis for most melanoma cells (Gal et al., 2015; Lee et al., 2019;
Piskounova et al., 2015). Ubellacker et al., (2020) recently investi-
gated this process and found that lymph specifically protects me-
tastasizing cells from ferroptosis. They obtained sets of efficiently
metastasizing and inefficiently metastasizing patient melanoma
samples as well as YUMM1.7 and YUMM3.3 syngeneic mouse mela-
noma models and tested their ability to form tumors when injected
 |
22       TALTY and BOSENBERG
into mice under different conditions. For all cell lines, the group no-
ticed that fewer cells were necessary to form tumors when injected
into lymph nodes than when injected intravenously. Melanoma cells
in the blood had greater ROS, lower ratios of glutathione to oxidized
glutathione, and increased ferroptosis. Furthermore, pretreatment
of cells with ferroptosis inhibitors increased intravenous metastasis
but not intranodal metastasis. Loss of GPX4, on the other hand, re-
duced the burden of tumors formed from intravenous injections but
not intranodal or subcutaneous ones. Ultimately, the group identi-
fied oleic acid as a lymph component that directly protects cells from
ferroptosis.
Sato et al. also interrogated the role of ferroptosis in melanoma
pathogenesis and metastasis by generating System Xc-­-­deficient
B16F10 melanoma cells (Sato et al., 2020). Loss of System Xc-­
these cells reduced cystine uptake, cellular glutathione, cell cycle
progression, and proliferation in vitro, tumor spheroid formation
ex vivo, and subcutaneous tumor formation in vivo. Importantly,
none of these changes could be rescued by the ferroptosis inhib-
itor liproxstatin-­1. System Xc-­-­deficient cells also adhered poorly
to lung vascular endothelium in vitro, showed reduced migration
potential by in vitro scratch assays, and formed less metastases
in vivo via tail vein, intrasplenic, IP, and footpad injections. These
studies collectively link ferroptosis susceptibility and metastatic
potential in melanoma. Notably, neither study directly evaluated
the roles of immune cells on metastasis. New evidence suggests
this might be especially important in the context of lymph metas-
tases, and it will be important to investigate this in the future (Song
et al., 2020).
7 |   I M M U N E C H EC K P O I NT I N H I B ITI O N
AC TI VATE S C D 8 + T C E LL S TO E N H A N C E
FE R RO P TOS I S I N T U M O R C E LL S
Inactivation of GPX4 in the brain and kidney led to the first experi-
mental hints that ferroptosis leads to a cellular immune response.
Neuron-­specific GPX4 loss induces strong immunoreactivity of
brain tissue as measured by the degree of reactive gliosis (Seiler
et al., 2008). In the kidney, loss of GPX4 causes release of damage-­
associated molecular patterns (DAMPs), recruitment of mac-
rophages, and massive tubular death and renal failure (Friedmann
Angeli et al., 2014). Given this, Wang et al., (2019) decided to in-
vestigate a potential role for ferroptosis in antitumor immunity
and cancer immunotherapy. First, they showed that treatment of
B16 melanoma xenografts with α-­PD-­1 immune checkpoint inhi-
bition elicits lipid peroxidation and ferroptosis in tumor cells but
not tumor-­infiltrating lymphocytes. They then demonstrated, first
by treatment of an erastin-­resistant cell line and later by coad-
ministration of the ferroptosis inhibitor liproxstatin-­1, that ferrop-
tosis is necessary for the antitumor efficacy of immunotherapy.
Mechanistically, they determined that IFN-­γ secreted by CD8+ T
cells reduces the expression of the System Xc-­ subunits SLC3A2 and
SLC7A11. This impairs cystine uptake by tumor cells, reduces the
synthesis of reduced glutathione, and promotes lipid peroxidation
and ferroptosis. They also evaluated whether expression of System
Xc-­ subunits in human melanoma tissues correlates with immuno-
therapy outcomes. Reduced expression of SLC3A2 correlated with a
strong CD8+ T cell signature, IFN-­γ expression, and improved over-
all patient survival.
More recently, Jiang, Lim, et al., (2021) showed that limiting tu-
moral ferroptosis via upregulation of TYRO3 promotes resistance to
α-­PD-­1/PD-­L1 immune checkpoint inhibition. They performed an
unbiased screen with receptor tyrosine kinase (RTK) antibodies that
identified elevated TYRO3 expression in α-­PD-­1-­resistant cells. They
then found in two publicly available gene expression databases,
Tumor Immune Dysfunction and Exclusion (TIDE) and Prediction
in of Clinical Outcomes from Genomics Profiles (PRECOG), that
higher TYRO3 expression correlated with shorter overall survival
in melanoma patients treated with α-­PD-­1 checkpoint inhibitors.
Overexpression of TYRO3 was sufficient to generate resistance to
α-­PD-­1 in mouse tumors, and knockout of TYRO3 in α-­PD-­1-­resistant
tumors conferred sensitivity to a-­PD-­1. In TYRO3-­overexpressing
cells, genes that block ferroptosis were upregulated (e.g., SLC40A1,
SLC7A11, SLC3A2, and GPX4), whereas genes that enhance ferro-
ptosis were downregulated (e.g., SLC5A1, TFRC). These cells were
also resistant to treatment with erastin in vitro, generated less lipid
peroxides when treated with α-­PD-­1 in vivo, and reduced the M1/
M2-­like macrophage ratio to create a protumor microenvironment.
Inhibition of TYRO3 promoted tumor ferroptosis and sensitized α-­
PD-­1-­resistant tumors to α-­PD-­1 treatment. Figure 2. compares nor-
mal CD8+ T cell–­tumor interaction after PD-­1 checkpoint blockade
with their interaction in the setting of TYRO3 overexpression by
tumor cells.
These reports highlight an exciting, new possibility for the im-
provement of existing immunotherapy strategies by modulating
ferroptosis. However, pathways that promote ferroptosis resistance
in tumors such as TYRO3 upregulation have already emerged, and
several unanswered questions remain. For example, what effect
does tumor cell ferroptosis have on immune cells other than CD8+
T cells and how does ferroptosis shape the overall tumor microenvi-
ronment? Jiang et al. showed that TYRO3-­overexpressing cells alter
macrophage polarization to favor immune suppression and tumor
growth, but it remains unclear whether this also occurs with other
types of ferroptosis-­resistant cells. The relative sensitivities of spe-
cific immune cell populations in the tumor microenvironment to
systemic ferroptosis inducers like RSL3 or imidazole ketone erastin
are also unknown. Retrospective clinical studies that examine the
effects of iron status, thiazolidine use, and statin use on immuno-
therapy outcomes in patients with metastatic melanoma could also
prove insightful. Statins sensitize cells to ferroptosis through their
modulation of mevalonate synthesis, whereas thiazolidines inhibit
the ferroptosis promoting enzyme ACSL4 and reduce lipid perox-
ide production. If ferroptosis is a key component of the antitumor
action of immune checkpoint inhibition, it is possible that targeting
specific ferroptosis regulatory pathways could enhance therapeutic
responses.
TALTY and BOSENBERG
F I G U R E 2   CD8+ T cell–­tumor
interaction after PD-­1 checkpoint
blockade. (a) Under normal conditions,
CD8+ T cells release IFN-­γ which causes
tumor cells to downregulate SLC3A2
and SLC7A11, depletes their cysteine
stores, and triggers ferroptotic cell death.
(b) Overexpression of TYRO3 by tumor
cells causes them to upregulate SLC3A2,
SLC7A11, and GPX4 which confer
resistance to ferroptotic cell death and
checkpoint blockade
8 |  M U LTI PLE PATH WAYS C A N CO N FE R
R E S I S TA N C E TO FE R RO P TOS I S I N
MEL ANOMA
Given the therapeutic potential of ferroptosis regulators in
melanoma, a better understanding of how resistance to these
compounds can develop will be critical for the development of ef-
fective clinical strategies. Two studies have explored resistance
to the ferroptosis inducer erastin. Although erastin is primarily
thought to induce ferroptosis by inhibiting System Xc-­, it also binds
to the voltage-­dependent anion channels VDAC2 and VDAC3 to
alter the permeability of the outer mitochondrial membrane and
decrease the rate of NADH oxidation. In the first study, the au-
thors observed a dramatic downregulation of VDAC2/3 in A375
melanoma cells just hours after treatment with erastin that con-
tributes to erastin resistance (Yang et al., 2020). They then showed
that the E3 ligase NEDD4 recognizes, binds to, ubiquinates, and
degrades VDAC2/3 under such conditions. Furthermore, deple-
tion of NEDD4 by shRNA blocked erastin-­induced degradation of
VDAC2/3 and increased the sensitivity of melanoma cells to fer-
roptosis. A second study focused on resistance to erastin in four
human melanoma cell lines (A375, CHL-­1, C8161, and SK-­Mel-­5)
(Gagliardi et al., 2019). They showed that most melanoma cell lines
resist ferroptosis execution through the NRF2-­driven upregulation
of aldo–­keto reductases (AKRs), which catalyze the conversion of
aldehydes and ketones to their corresponding alcohols to help re-
duce lipid peroxide accumulation. The relevance of this pathway
in vivo, however, is less clear as they found no associations be-
tween tumor NRF2 or AKR expression and survival in patients with
metastatic melanoma. They did, however, identify SLC7A11 as a
useful marker to predict the susceptibility of metastatic melanoma
cells to ferroptosis. This specifically highlights the need for future
|
      23
preclinical studies to directly examine the role of ferroptosis in
models of melanoma metastasis.
9 | CO N C LU S I O N A N D FU T U R E
D I R EC TI O N S
Although ferroptosis was only recently discovered as a distinct
mechanism of regulated cell death, accumulating evidence sug-
gests that regulation of ferroptosis plays multiple roles in melanoma
progression and therapeutic resistance. Multiple lines of evidence
support a role for ferroptotic pathways regulating the differentia-
tion state of melanoma cells and resistance to particular therapeutic
agents. It is clear that ferroptosis inducers have strong therapeutic
potential in melanoma—­both as single agents to overcome resistance
to conventional chemotherapy or targeted therapy and in combina-
tion with targeted therapies or immune checkpoint inhibitors to aug-
ment antitumor immunity and increase the overall patient response
rates. The continued development of novel ferroptosis inducers
with improved pharmacokinetics such as engineered cyst(e)inase,
imidazole ketone erastin, and JKE-­1674 will further broaden thera-
peutic opportunities (Cramer et al., 2017; Eaton et al., 2020; Zhang
et al., 2019). Several pathways of resistance to ferroptosis inducers
in melanoma, however, have already been identified and will be im-
portant to keep in mind as clinical trials for ferroptosis inducers are
designed. In the next several years, research on ferroptosis in mela-
noma will undoubtedly continue to expand and will likely delineate
the roles of additional potential regulators of ferroptosis, including
iron metabolism, transsulfuration, hippo signaling, and mevalonate
synthesis pathways. How current findings replicate across multiple
melanoma models and how well they translate to human patients
with melanoma represent critical outstanding questions.
 |
24       TALTY and BOSENBERG
AC K N OW L E D G E M E N T S
This work was funded by grants to Marcus Bosenberg and Ronan
Talty (NIH F30CA254246-­01A1) from the National Cancer Institute
of the National Institute of Health. The content is solely the respon-
sibility of the authors and does not necessarily represent the offi-
cial views of funding agencies. The figures in this work were created
with BioRender.
C O N FL I C T S O F I N T E R E S T
The authors declare no conflicts of interest.
ORCID
Ronan Talty  https://orcid.org/0000-0002-8742-3799
REFERENCES
Bersuker, K., Hendricks, J. M., Li, Z., Magtanong, L., Ford, B., Tang, P.
H., Roberts, M. A., Tong, B., Maimone, T. J., Zoncu, R., Bassik, M.
C., Nomura, D. K., Dixon, S. J., & Olzmann, J. A. (2019). The CoQ
oxidoreductase FSP1 acts parallel to GPX4 to inhibit ferroptosis.
Nature, 575, 688–­692. https://doi.org/10.1038/s4158​6 - ­019-­1705-­2
Boumahdi, S., & de Sauvage, F. J. (2020). The great escape: Tumour cell
plasticity in resistance to targeted therapy. Nat. Rev. Drug Discov., 19,
39–­56. https://doi.org/10.1038/s4157​3- ­019- ­0 044-­1
Brown, C. W., Amante, J. J., Chhoy, P., Elaimy, A. L., Liu, H., Zhu, L. J.,
Baer, C. E., Dixon, S. J., & Mercurio, A. M. (2019). Prominin2 drives
ferroptosis resistance by stimulating iron export. Developmental Cell,
51, 575–­586.e4. https://doi.org/10.1016/j.devcel.2019.10.007
Conrad, M., & Pratt, D. A. (2019). The chemical basis of ferroptosis.
Nature Chemical Biology, 15, 1137–­ 1147. https://doi.org/10.1038/
s4158​9-­019-­0 408-­1
Cramer, S. L., Saha, A., Liu, J., Tadi, S., Tiziani, S., Yan, W., Triplett, K.,
Lamb, C., Alters, S. E., Rowlinson, S., Zhang, Y. J., Keating, M. J.,
Huang, P., DiGiovanni, J., Georgiou, G., & Stone, E. (2017). Systemic
depletion of L-­c yst(e)ine with cyst(e)inase increases reactive oxygen
species and suppresses tumor growth. Nature Medicine, 23, 120–­127.
https://doi.org/10.1038/nm.4232
Dixon, S. J., Lemberg, K. M., Lamprecht, M. R., Skouta, R., Zaitsev, E.
M., Gleason, C. E., Patel, D. N., Bauer, A. J., Cantley, A. M., Yang,
W. S., Morrison, B., & Stockwell, B. R. (2012). Ferroptosis: An iron-­
dependent form of nonapoptotic cell death. Cell, 149, 1060–­1072.
https://doi.org/10.1016/j.cell.2012.03.042
Dodson, M., Castro-­Portuguez, R., & Zhang, D. D. (2019). NRF2 plays
a critical role in mitigating lipid peroxidation and ferroptosis. Redox
Biology, 23. https://doi.org/10.1016/j.redox.2019.101107
Doll, S., Freitas, F. P., Shah, R., Aldrovandi, M., da Silva, M. C., Ingold,
I., Grocin, A. G., Xavier da Silva, T. N., Panzilius, E., Scheel, C. H.,
Mourão, A., Buday, K., Sato, M., Wanninger, J., Vignane, T., Mohana,
V., Rehberg, M., Flatley, A., Schepers, A. …Conrad, M. (2019). FSP1 is
a glutathione-­independent ferroptosis suppressor. Nature, 575, 693–­
698. https://doi.org/10.1038/s4158​6-­019-­1707-­0
Dolma, S., Lessnick, S. L., Hahn, W. C., & Stockwell, B. R. (2003).
Identification of genotype-­selective antitumor agents using synthetic
lethal chemical screening in engineered human tumor cells. Cancer
Cell, 3, 285–­296. https://doi.org/10.1016/S1535​-­6108(03)00050​-­3
Eaton, J. K., Furst, L., Ruberto, R. A., Moosmayer, D., Hilpmann, A., Ryan,
M. J., Zimmermann, K., Cai, L. L., Niehues, M., Badock, V., Kramm,
A., Chen, S., Hillig, R. C., Clemons, P. A., Gradl, S., Montagnon, C.,
Lazarski, K. E., Christian, S., Bajrami, B. …Schreiber, S. L. (2020).
Selective covalent targeting of GPX4 using masked nitrile-­ oxide
electrophiles. Nature Chemical Biology, 16, 497–­ 506. https://doi.
org/10.1038/s4158​9-­020-­0501-­5
Friedmann Angeli, J. P., Krysko, D. V., & Conrad, M. (2019). Ferroptosis at
the crossroads of cancer-­acquired drug resistance and immune eva-
sion. Nature Reviews Cancer, 19.
Friedmann Angeli, J. P., Schneider, M., Proneth, B., Tyurina, Y. Y.,
Tyurin, V. A., Hammond, V. J., Herbach, N., Aichler, M., Walch, A.,
Eggenhofer, E., Basavarajappa, D., Rådmark, O., Kobayashi, S., Seibt,
T., Beck, H., Neff, F., Esposito, I., Wanke, R., Förster, H. … Conrad, M.
(2014). Inactivation of the ferroptosis regulator Gpx4 triggers acute
renal failure in mice. Nature Cell Biology, 16, 1180–­1191.
Gagliardi, M., Cotella, D., Santoro, C., Corà, D., Barlev, N. A., Piacentini,
M., & Corazzari, M. (2019). Aldo-­keto reductases protect metastatic
melanoma from ER stress-­ independent ferroptosis. Cell Death &
Disease, 10, 902. https://doi.org/10.1038/s4141​9-­019-­2143-­7
Gal, K. L., Ibrahim, M. X., Wiel, C., Sayin, V. I., Akula, M. K., Karlsson,
C., Dalin, M. G., Akyürek, L. M., Lindahl, P., Nilsson, J. et al (2015).
Antioxidants can increase melanoma metastasis in mice. Science
Translational Medicine, 7, 308re8-­3 08re8.
Gao, M., Monian, P., Quadri, N., Ramasamy, R., & Jiang, X. (2015).
Glutaminolysis and transferrin regulate ferroptosis. Molecular Cell,
59, 298–­3 08. https://doi.org/10.1016/j.molcel.2015.06.011
Gupta, P. B., Pastushenko, I., Skibinski, A., Blanpain, C., & Kuperwasser,
C. (2019). Phenotypic plasticity: Driver of cancer initiation, progres-
sion, and therapy resistance. Cell Stem Cell, 24, 65–­78. https://doi.
org/10.1016/j.stem.2018.11.011
Hangauer, M. J., Viswanathan, V. S., Ryan, M. J., Bole, D., Eaton, J. K.,
Matov, A., Galeas, J., Dhruv, H. D., Berens, M. E., Schreiber, S. L.,
McCormick, F., & McManus, M. T. (2017). Drug-­tolerant persister
cancer cells are vulnerable to GPX4 inhibition. Nature, 551, 247–­250.
https://doi.org/10.1038/natur​e24297
Hayano, M., Yang, W. S., Corn, C. K., Pagano, N. C., & Stockwell, B. R.
(2016). Loss of cysteinyl-­tRNA synthetase (CARS) induces the trans-
sulfuration pathway and inhibits ferroptosis induced by cystine
deprivation. Cell Death and Differentiation, 23, 270–­278. https://doi.
org/10.1038/cdd.2015.93
Hugo, W., Shi, H., Sun, L., Piva, M., Song, C., Kong, X., Moriceau, G., Hong,
A., Dahlman, K. B., Johnson, D. B., Sosman, J. A., Ribas, A., & Lo, R.
S. (2015). Non-­genomic and immune evolution of melanoma acquiring
MAPKi resistance. Cell, 162, 1271–­ 1285. https://doi.org/10.1016/j.
cell.2015.07.061
Jiang, X., Stockwell, B. R., & Conrad, M. (2021). Ferroptosis: Mechanisms,
biology and role in disease. Nature Reviews Molecular Cell Biology, 22,
266–­282. https://doi.org/10.1038/s4158​0 -­020-­0 0324​-­8
Jiang, Z., Lim, S. O., Yan, M., Hsu, J. L., Yao, J., Wei, Y., Chang, S. S.,
Yamaguchi, H., Lee, H. H., Ke, B., Hsu, J.-­M., Chan, L.-­C ., Hortobagyi,
G. N., Yang, L., Lin, C., Yu, D., & Hung, M.-­C . (2021). TYRO3 induces
anti–­PD-­ 1/PD-­L1 therapy resistance by limiting innate immunity
and tumoral ferroptosis. J Clin Invest, 131(8), e139434. https://doi.
org/10.1172/JCI13​9434
Johnson, W. M., Wilson-­ Delfosse, A. L., & Mieyal, J. J. (2012).
Dysregulation of glutathione homeostasis in neurodegenerative dis-
eases. Nutrients, 4, 1399–­1440. https://doi.org/10.3390/nu410​1399
Lee, C.-­K ., Jeong, S.-­H., Jang, C., Bae, H., Kim, Y. H., Park, I., Kim, S.
K., & Koh, G. Y. (2019). Tumor metastasis to lymph nodes requires
YAP-­dependent metabolic adaptation. Science (80-­.). 363, 644–­6 49.
https://doi.org/10.1126/scien​ce.aav0173
Lu, B., Chen, X. B., Ying, M. D., He, Q. J., Cao, J., & Yang, B. (2017). The
role of ferroptosis in cancer development and treatment response.
Front. Pharmacol, 8, 992. https://doi.org/10.3389/fphar.2017.00992
Luo, M., Wu, L., Zhang, K., Wang, H., Zhang, T., Gutierrez, L., O’Connell,
D., Zhang, P., Li, Y., Gao, T., Ren, W., & Yang, Y. (2018). miR-­137 reg-
ulates ferroptosis by targeting glutamine transporter SLC1A5 in
melanoma. Cell Death and Differentiation, 25, 1457–­1472. https://doi.
org/10.1038/s4141​8-­017-­0 053-­8
Masaldan, S., Bush, A. I., Devos, D., Rolland, A. S., & Moreau, C. (2019).
Striking while the iron is hot: Iron metabolism and ferroptosis in
TALTY and BOSENBERG
neurodegeneration. Free Radical Biology and Medicine, 133, 221–­233.
https://doi.org/10.1016/j.freer​adbio​med.2018.09.033
Müller, J., Krijgsman, O., Tsoi, J., Robert, L., Hugo, W., Song, C., Kong, X.,
Possik, P. A., Cornelissen-­Steijger, P. D. M., Foppen, M. H. G., Kemper,
K., Goding, C. R., McDermott, U., Blank, C., Haanen, J., Graeber, T. G.,
Ribas, A., Lo, R. S., & Peeper, D. S. (2014). Low MITF/AXL ratio pre-
dicts early resistance to multiple targeted drugs in melanoma. Nature
Communications, 5. https://doi.org/10.1038/ncomm​s6712
Murphy, T. H., Miyamoto, M., Sastre, A., Schnaar, R. L., & Coyle, J. T.
(1989). Glutamate toxicity in a neuronal cell line involves inhibition of
cystine transport leading to oxidative stress. Neuron, 2, 1547–­1558.
https://doi.org/10.1016/0896-­6273(89)90043​-­3
Piskounova, E., Agathocleous, M., Murphy, M. M., Hu, Z., Huddlestun,
S. E., Zhao, Z., Leitch, A. M., Johnson, T. M., DeBerardinis, R. J., &
Morrison, S. J. (2015). Oxidative stress inhibits distant metasta-
sis by human melanoma cells. Nature, 527, 186–­ 191. https://doi.
org/10.1038/natur​e15726
Riesenberg, S., Groetchen, A., Siddaway, R., Bald, T., Reinhardt, J.,
Smorra, D., Kohlmeyer, J., Renn, M., Phung, B., Aymans, P., Schmidt,
T., Hornung, V., Davidson, I., Goding, C. R., Jönsson, G., Landsberg, J.,
Tüting, T., & Hölzel, M. (2015). MITF and c-­Jun antagonism intercon-
nects melanoma dedifferentiation with pro-­inflammatory cytokine
responsiveness and myeloid cell recruitment. Nature Communications,
6. https://doi.org/10.1038/ncomm​s9755
Robert, C., Grob, J. J., Stroyakovskiy, D., Karaszewska, B., Hauschild,
A., Levchenko, E., Chiarion Sileni, V., Schachter, J., Garbe, C.,
Bondarenko, I., Gogas, H., Mandalá, M., Haanen, J. B. A. G., Lebbé,
C., Mackiewicz, A., Rutkowski, P., Nathan, P. D., Ribas, A., Davies,
M. A. …Long, G. V. (2019). Five-­year outcomes with dabrafenib plus
trametinib in metastatic melanoma. New England Journal of Medicine,
381, 626–­636. https://doi.org/10.1056/NEJMo​a1904059
Sato, M., Onuma, K., Domon, M., Hasegawa, S., Suzuki, A., Kusumi, R., Hino,
R., Kakihara, N., Kanda, Y., Osaki, M., Hamada, J., Bannai, S., Feederle,
R., Buday, K., Angeli, J. P. F., Proneth, B., Conrad, M., Okada, F., & Sato,
H. (2020). Loss of the cystine/glutamate antiporter in melanoma abro-
gates tumor metastasis and markedly increases survival rates of mice.
International Journal of Cancer. https://doi.org/10.1002/ijc.33262
Sauka-­Spengler, T., & Bronner-­Fraser, M. (2008). A gene regulatory net-
work orchestrates neural crest formation. Nature Reviews Molecular
Cell Biology, 9, 557–­568. https://doi.org/10.1038/nrm2428
Schachter, J., Ribas, A., Long, G. V., Arance, A., Grob, J. J., Mortier, L.,
Daud, A., Carlino, M. S., McNeil, C., Lotem, M., Larkin, J., Lorigan, P.,
Neyns, B., Blank, C., Petrella, T. M., Hamid, O., Zhou, H., Ebbinghaus,
S., Ibrahim, N., & Robert, C. (2017). Pembrolizumab versus ipilimumab
for advanced melanoma: Final overall survival results of a multicentre,
randomised, open-­label phase 3 study (KEYNOTE-­0 06). Lancet, 390,
1853–­1862. https://doi.org/10.1016/S0140​-­6736(17)31601​-­X
Seiler, A., Schneider, M., Förster, H., Roth, S., Wirth, E. K., Culmsee, C.,
Plesnila, N., Kremmer, E., Rådmark, O., Wurst, W., Bornkamm, G. W.,
Schweizer, U., & Conrad, M. (2008). Glutathione peroxidase 4 senses
and translates oxidative stress into 12/15-­lipoxygenase dependent-­
and AIF-­mediated cell death. Cell Metabolism, 8, 237–­248. https://
doi.org/10.1016/j.cmet.2008.07.005
Siegel, R. L., Miller, K. D., & Jemal, A. (2019). Cancer statistics, 2019. CA:
A Cancer Journal for Clinicians, 69, 7–­3 4. https://doi.org/10.3322/
caac.21551
Song, E., Mao, T., Dong, H., Boisserand, L. S. B., Antila, S., Bosenberg,
M., Alitalo, K., Thomas, J. L., & Iwasaki, A. (2020). VEGF-­C-­driven
lymphatic drainage enables immunosurveillance of brain tumours.
Nature, 577, 689–­694. https://doi.org/10.1038/s4158​6-­019-­1912-­x
Stockwell, B. R., & Jiang, X. (2020). The chemistry and biology of ferro-
ptosis. Cell Chem Biol, 27, 365–­375. https://doi.org/10.1016/j.chemb​
iol.2020.03.013
Tsoi, J., Robert, L., Paraiso, K., Galvan, C., Sheu, K. M., Lay, J., Wong, D.
J. L., Atefi, M., Shirazi, R., Wang, X., Braas, D., Grasso, C. S., Palaskas,
N., Ribas, A., & Graeber, T. G. (2018a). Multi-­stage differentiation
|
      25
defines melanoma subtypes with differential vulnerability to drug-­
induced iron-­dependent oxidative stress. Cancer Cell, 33, 890–­904.
e5. https://doi.org/10.1016/j.ccell.2018.03.017
Tsoi, J., Robert, L., Paraiso, K., Galvan, C., Sheu, K. M., Lay, J., Wong, D.
J. L., Atefi, M., Shirazi, R., Wang, X., Braas, D., Grasso, C. S., Palaskas,
N., Ribas, A., & Graeber, T. G. (2018b). Multi-­stage differentiation
defines melanoma subtypes with differential vulnerability to drug-­
induced iron-­dependent oxidative stress. Cancer Cell, 33, 890–­904.
e5. https://doi.org/10.1016/j.ccell.2018.03.017
Ubellacker, J. M., Tasdogan, A., Ramesh, V., Shen, B., Mitchell, E. C.,
Martin-­Sandoval, M. S., Gu, Z., McCormick, M. L., Durham, A. B.,
Spitz, D. R., Zhao, Z., Mathews, T. P., & Morrison, S. J. (2020). Lymph
protects metastasizing melanoma cells from ferroptosis. Nature, 585,
113. https://doi.org/10.1038/s4158​6-­020-­2623-­z
Viswanathan, V. S., Ryan, M. J., Dhruv, H. D., Gill, S., Eichhoff, O. M.,
Seashore-­Ludlow, B., Kaffenberger, S. D., Eaton, J. K., Shimada, K.,
Aguirre, A. J., Viswanathan, S. R., Chattopadhyay, S., Tamayo, P.,
Yang, W. S., Rees, M. G., Chen, S., Boskovic, Z. V., Javaid, S., Huang,
C. …Schreiber, S. L. (2017). Dependency of a therapy-­resistant state
of cancer cells on a lipid peroxidase pathway. Nature, 547, 453–­457.
https://doi.org/10.1038/natur​e23007
Wang, W., Green, M., Choi, J. E., Gijón, M., Kennedy, P. D., Johnson, J.
K., Liao, P., Lang, X., Kryczek, I., Sell, A., Xia, H., Zhou, J., Li, G., Li,
J., Li, W., Wei, S., Vatan, L., Zhang, H., Szeliga, W. … Zou, W. (2019).
CD8+ T cells regulate tumour ferroptosis during cancer immuno-
therapy. Nature, 569, 270–­ 274. https://doi.org/10.1038/s4158​
6-­019-­1170-­y
Warner, G. J., Berry, M. J., Moustafa, M. E., Carlson, B. A., Hatfield, D. L.,
& Faust, J. R. (2000). Inhibition of selenoprotein synthesis by sele-
nocysteine tRNA([Ser]Sec) lacking isopentenyladenosine. Journal of
Biological Chemistry, 275, 28110–­28119. https://doi.org/10.1074/jbc.
M0012​8 0200
Wilson, M. A., Zhong, J., Rosenbaum, B. E., Utter, K., Moran, U.,
Darvishian, F., Polsky, D., Berman, R. S., Shapiro, R. L., Pavlick, A. C.,
& Osman, I. (2019). Impact of initial stage on metastatic melanoma
survival. Melanoma Research, 29, 281–­288. https://doi.org/10.1097/
CMR.00000​0 0000​0 00526
Wu, J., Minikes, A. M., Gao, M., Bian, H., Li, Y., Stockwell, B. R., Chen,
Z. N., & Jiang, X. (2019). Intercellular interaction dictates cancer cell
ferroptosis via NF2–­YAP signalling. Nature, 572, 402–­4 06. https://
doi.org/10.1038/s4158​6-­019-­1426-­6
Yang, W. S., & Stockwell, B. R. (2008). Synthetic lethal screening identi-
fies compounds activating iron-­dependent, nonapoptotic cell death
in oncogenic-­R AS-­harboring cancer cells. Chemistry & Biology, 15,
234–­245. https://doi.org/10.1016/j.chemb​iol.2008.02.010
Yang, Y., Luo, M., Zhang, K., Zhang, J., Gao, T., Connell, D. O., Yao, F.,
Mu, C., Cai, B., Shang, Y. & Chen, W. (2020). Nedd4 ubiquitylates
VDAC2/3 to suppress erastin-­ induced ferroptosis in melanoma.
Nature Communications, 11. https://doi.org/10.1038/s4146​7-­020-­
14324​-­x
Zhang, K., Wu, L., Zhang, P., Luo, M., Du, J., Gao, T., O’Connell, D., Wang,
G., Wang, H., & Yang, Y. (2018). miR-­9 regulates ferroptosis by target-
ing glutamic-­oxaloacetic transaminase GOT1 in melanoma. Molecular
Carcinogenesis, 57, 1566–­1576. https://doi.org/10.1002/mc.22878
Zhang, Y., Tan, H., Daniels, J. D., Zandkarimi, F., Liu, H., Brown, L. M.,
Uchida, K., O’Connor, O. A., & Stockwell, B. R. (2019). Imidazole ke-
tone erastin induces ferroptosis and slows tumor growth in a mouse
lymphoma model. Cell Chem. Biol., 26, 623–­ 633.e9. https://doi.
org/10.1016/j.chemb​iol.2019.01.008
How to cite this article: Talty, R., & Bosenberg, M. (2022).
The role of ferroptosis in melanoma. Pigment Cell & Melanoma
Research, 35, 18–­25. https://doi.org/10.1111/pcmr.13009