Ferroptosis 표적화

https://www.sciencedirect.com/science/article/pii/S1535610819301977

Targeting Ferroptosis to Iron Out Cancer

2019

Table 1. Ferroptosis Stimuli

MechanismTarget            Compound/Drug                                          Phenotype and Readout

GPX4 inactivation due to GSH depletion (class I FINs)	system XC−	erastin	lipid ROSa death inhibited by DFO, Fer1, and Trolox
		piperazine erastin	increase in PTGS2 mRNA regression HT1080 xenografts
		imidazole ketone erastin (IKE)	lipid ROSa
		sulfasalazine	death inhibited by DFO, Fer1, and Trolox
		sorafenib	lipid ROSa death inhibited by DFO and Fer1
		glutamate	inhibited by CPX and Fer1
	glutamate-cysteine ligase	buthionine sulfoximine (BSO)	lipid ROSa
	glutathione S-transferase	artesunate	lipid ROSa death inhibited by DFO, Fer1, and Trolox
	unknown	DPI2	death inhibited by DFO and α-Toc
	[Cys] depletion	cyst(e)inase	reduced GSH levels regression of MDA-MB-361, DU145 and PC3 xenografts
	ND dehydrogenase	BAY 87-2243	lipid ROSa death inhibited by Fer1 and α-Toc
GPX4 inactivation/depletion (class II, III FINs)	GPX4	1S,3R-RSL3b,c	lipid ROSa death inhibited by DFO and Fer1 increase in PTGS2 mRNA HT1080 tumor regression
		DPI7/ML162b,DPI10/ML210b, DPI12-13b, DPI17-19b	lipid ROSa
		altretamineb	lipid ROSa
		FIN56b	lipid ROSa death inhibited by DFO and α-Toc
		withaferin Ac	lipid ROSa in neuroblastoma cells death inhibited by DFO, Fer1 and α-Toc
	Squalene synthase	FIN56b	lipid ROSa death inhibited by DFO and α-Toc
	HMG-CoA reductase	fluvastatin, lovastatin acid, simvastatin	lipid ROSa
Iron loading (class IV FINs)		hemoglobin	lipid ROSa death inhibited by Fer1 in organotypic hippocampal slice cultures
		FeCl2	death inhibited by Fer1 in organotypic hippocampal slice cultures
		hemin	death inhibited by DFO and Fer1
		(NH4)2Fe(SO4)2	death inhibited by Fer1
		non-thermal plasma	lipid ROSa death inhibited by DFO
		salinomycin	lipid ROSa death inhibited by Fer1
		amino acid depletion + Cornell dots	lipid ROSa death inhibited by Fer1 and Trolox
		amino acid depletion, cystine deprivation + holo-transferrin	death inhibited by Fer1
	transferrin ↑ ferroportin-1 ↓	lapatinib + siramesine	death inhibited by DFO and Fer1
Iron oxidation (class IV FINs)		FINO2	lipid ROSa death inhibited by Fer1
Increase in LIP by HMOX1↑ (class IV FINs)	KEAP1 inactivation	withaferin A	lipid ROSa in neuroblastoma death inhibited by DFO, Fer1 and α-Toc
	IκBα	BAY 11-7085	death inhibited by Fer1, Lip1 and NAC
Unknown		lanperisone	death inhibited by DFO and Trolox
		artemisinin derivatives	death inhibited by DFO and Fer1
		CIL41, CIL56, CIL69, CIL70, CIL75, and CIL79	lipid ROSa for CIL56 death inhibited by DFO and α-Toc

α-Toc, α-tocopherol; CIL, caspase-3/7-independent lethal; CPX, ciclopirox olamine; DFO, deferoxamine; Fer1, ferrostatin-1; FIN, ferroptosis-inducing compound, GPX4, glutathione peroxidase 4; GSH, glutathione; HMOX1, heme oxygenase-1; IκBα, nuclear factor of κ light-chain polypeptide gene enhancer in B cell inhibitor α; KEAP1, kelch-like ECH-associated protein 1; LIP, labile iron pool; Lip1, liproxstatin-1; PTGS2, prostaglandin-endoperoxide synthase 2, ROS, reactive oxygen species; VDAC, voltage-dependent anion channel.

a

Lipid ROS shown by C11-BODIPY staining.

b

GPX4 inactivation shown using LC-MS-based GPX4 assay (PCOOH).

c

Direct GPX4 binding shown through pull-down.

 

 

그림 2 . Ferroptosis 유도의 메커니즘

 

(A) 클래스 I 페로프토시스 유도 화합물(FIN) 은 시스템 X C -의 억제 또는 글루타메이트-시스테인 리가제(GCL)의 억제를 통해 세포내 글루타티온 (GSH)을 고갈시킴으로써 페로프토시스를 유발 합니다.

 

(B) 클래스 II FIN은 GPX4의 직접적인 억제 및 불활성화에 의해 페로프토시스를 유발 합니다.

 

(C) 클래스 III FIN은 GPX4의 간접적인 억제 및 비활성화에 의해 페로프토시스를 유발합니다. 

예를 들어, FIN56 은 콜레스테롤 합성에 관여하는 효소인 스쿠알렌 합성효소 (SQS)에 결합하여 활성화 합니다. 이것은 CoQ 10 및 Sec-tRNA와 같은 일부 대사 산물의 억제를 초래할 수 있으며 GPX4의 관찰된 고갈/불활성화를 설명합니다.

 

(D) 클래스 IV FIN은 직접적인 철 과부하 또는 헴 옥시게나제 1 (HMOX1) 의 과도한 활성을 통해 불안정한 철 풀(LIP)을 증가시켜 페로프토시스를 유발합니다. 

 

Glu, glutamate; BSO, buthionine sulfoximine; GR, glutathione reductase; GS, glutathione synthase; Gly, glycine, GSSG, glutathione disulfide; HMGCR, 3-hydroxy-3-methylglutaryl CoA reductase; WA, withaferin A; SAS, sulfasalazine; FPP, farnesyl pyrophosphate; KEAP1, kelch-like ECH-associated protein 1; NRF2, nuclear factor erythroid 2-related factor 2; DMT1, divalent metal transporter 1.

 

Table 2. Modulators of Ferroptosis

GeneProtein         Function             Modulatory Effect                                      References

TFRC	transferrin receptor	cellular transferrin-iron uptake	knockdown suppresses erastin-induced ferroptosis
			knockdown suppresses ferroptosis induced by amino acid/cystine deprivation
PHKG2	phosphorylase kinase, γ2	activates glycogen phosphorylase to release glucose-1-phosphate from glycogen	iron regulatory function? knockdown suppresses erastin-induced ferroptosis
IREB2	iron response element-binding protein 2	RNA-binding protein that regulates iron levels in the cells by regulating the translation and stability of mRNAs that affect iron homeostasis upon iron depletion	knockdown suppresses ferroptosis induced by erastin or amino acid/cystine deprivation
HSBP1	heat-shock 27-kDa protein 1	activated in heat stress response by heat-shock factor 1 (HSF1)	iron regulatory function? knockdown enhances erastin-induced ferroptosis in vitro and in vivo
HMOX1	heme oxygenase 1	catalyzes the degradation of heme to biliverdin, CO, and Fe2+	inhibition or knockdown suppresses withaferin A-induced ferroptosis
			inhibition or knockout suppresses erastin-induced ferroptosis
			inhibition or knockdown suppresses BAY-induced ferroptosis
CISD1/mitoNEET	CDGSH iron-sulfur domain 1	inhibits mitochondrial iron transport into the matrix	knockdown enhances erastin-induced ferroptosis
NCOA4	nuclear receptor coactivator 4	cargo receptor mediating ferritinophagy	knockdown suppresses ferroptosis induced by amino acid/cystine deprivation
			knockdown suppresses erastin-induced ferroptosis
ACO1	aconitase 1	iron-sulfur protein that converses citrate to isocitrate, controls iron inside cell	knockdown suppresses ferroptosis induced by amino acid/cystine deprivation
FTH1	ferritin heavy chain 1	subunit of major intracellular iron storage protein	expression level controls ferroptosis sensitivity
			knockdown enhances erastin- or sorafenib-induced ferroptosis in hepatocellular carcinoma
STEAP3	six-transmembrane epithelial antigen of prostate 3	metalloreductase converting Fe3+ to Fe2+	upregulated in response to erastin in bone marrow stromal cells
FANCD2	Fanconi anemia complementation group D2	nuclear protein involved in DNA damage repair	iron regulatory function? knockout enhances erastin-induced ferroptosis in bone marrow stromal cells
NFS1	cysteine desulfurase	enzyme involved in synthesizing iron-sulfur clusters using sulfur from cysteine	knockdown activates the iron-starvation response promoting erastin-induced ferroptosis
ACSF2	acyl-CoA synthetase family member 2	regulation of mitochondrial fatty acid metabolism	knockdown suppresses erastin-induced ferroptosis
CS	citrate synthase	regulation of mitochondrial fatty acid metabolism	knockdown suppresses erastin-induced ferroptosis
LPCAT3	lysophosphatidylcholine acyltransferase 3	incorporation of acylated fatty acids into membranes by catalyzing the reacylation of lysophospholipids to phospholipids	identified in haploid cell genetic screen
			knockdown suppresses RSL3-induced ferroptosis
ACSL4	acyl-CoA synthetase long-chain family member 4	converts free fatty acids (preferentially AA) into fatty acyl-CoAs	identified in haploid cell genetic screen
			knockdown suppresses erastin-induced ferroptosis
			inhibition or knockout suppresses RSL3-induced ferroptosis
			inhibition or knockout suppresses erastin-, RSL3-, or GPX4-depletion-induced ferroptosis inhibition suppresses GPX4-depletion-induced damage in vivo
ACSL3	acyl-CoA synthetase long-chain family member 3	converts exogenous monounsaturated fatty acids (MUFAs) into fatty acyl-CoAs	required for MUFA-induced protection from erastin2-induced ferroptosis knockout attenuates MUFA-induced resistance to ferroptosis
ACACA	Acetyl-CoA carboxylase alpha	Converts acetyl-CoA to malonyl-CoA, the rate-limiting step in fatty acid synthesis	identified in haploid cell genetic screen
			inhibition suppresses FIN56-, but not erastin- or RSL3- induced ferroptosis
GPX4	glutathione peroxidase 4	lipid repair enzyme	inhibition or knockout induces ferroptosis
AKR1C	aldo-keto reductase family 1 member C1	regulate the detoxification of oxidative lipid breakdown products	upregulation confers protection against erastin-induced ferroptosis
LOX	lipoxygenase-12/15	catalyzes the dioxygenation of polyunsaturated fatty acids in lipids	inhibition or knockout suppresses GPX4-depletion-induced ferroptosis
	lipoxygenases		knockout protects against imidazole keto erastin (IKE)-, but not RSL3-induced ferroptosis inhibition protects against erastin-induced ferroptosis
PEBP1	phosphatidylethanolamine-binding protein 1	protein scaffold	controls substrate specificity of LOX15 knockdown suppresses RSL3-induced ferroptosis
ZEB1	zinc finger E-box-binding homeobox 1	EMT regulator and lipogenic factor	knockout suppresses GPX4-depletion-induced ferroptosis
SQS/FDFT1	squalene synthase/farnesyl-diphosphate farnesyltransferase 1	responsible for synthesis of squalene and involved in cholesterol synthesis	knockdown or inhibition suppresses FIN-56-induced ferroptosis
			knockout or inhibition sensitizes SQLE-deficient anaplastic large cell lymphoma (ALCL) cells to ML162-induced ferroptosis
SQLE	squalene monooxygenase	catalyzes the conversion of squalene to squalene-2,3-epoxide and involved in cholesterol synthesis	overexpression sensitizes SQLE-deficient ALCL cells to ML162-induced ferroptosis
HMGCR	3-hydroxy-3-methyl-glutaryl-coenzyme A reductase	synthesis of mevalonic acid	inhibition enhances FIN-56-induced ferroptosis
FADS2	fatty acid desaturase 2/acyl-CoA 6-desaturase	involved in biosynthesis of highly unsaturated fatty acids, desaturates palmitate to produce the monounsaturated fatty acid sapienate	knockdown decreases sapienate production and suppresses RSL3- induced lipid peroxidation in HUH7 liver cancer cells
NRF2	nuclear factor erythroid 2-related factor 2	key regulator of anti-oxidant response including the expression of system XC−	inhibition or knockdown enhances erastin- or sorafenib-induced ferroptosis in hepatocellular carcinoma in vitro and in vivo
			knockdown enhances artesunate-induced ferroptosis in head and neck cancer in vitro and in vivo
			overexpression confers resistance to erastin- and RSL3-induced ferroptosis in glioma cells, while knockdown enhances ferroptosis
KEAP1	kelch-like ECH- associated protein 1	binds to and regulates NRF2 by keeping its levels at control	overexpression enhances erastin- and RSL3-induced ferroptosis in glioma cells, while knockdown confers resistance to ferroptosis
			knockdown confers resistance to artesunate-induced ferroptosis in head and neck cancer
HMOX1	heme oxygenase 1	catalyzes the degradation of heme to biliverdin, CO, and Fe2+	knockout enhances erastin-induced ferroptosis in proximal tubular cells
			knockout enhances erastin- and sorafenib-induced ferroptosis in hepatocellular carcinoma cells
NQO1	quinone oxidoreductase-1	reduces quinones to hydroquinones	knockdown enhances erastin- and sorafenib-induced ferroptosis in hepatocellular carcinoma cells
SLC7A11	solute carrier family 7 member 11	subunit of system Xc− to import cystine in the cell	inhibition induces ferroptosis
GCLC	glutamate-cysteine ligase catalytic subunit	enzyme involved in GSH synthesis	inhibition induces ferroptosis
CARS	cysteinyl-tRNA synthetase	protein translation	knockdown suppresses erastin-induced ferroptosis through compensatory activation of transsulfuration pathway [Cys]↑
CBS	cystathionine-β-synthase	converts homocysteine to cystathionine	inhibition or knockdown resensitizes erastin-induced ferroptosis in CARS knockdown cells
NOX	NADPH oxidase	superoxide-producing enzymes	inhibition suppresses erastin-induced ferroptosis in non-small cell lung cancer cells
ABCC1/MRP1	multidrug-resistance protein 1	mediates GSH and chemotherapeutics efflux from cells	overexpression sensitizes to ferroptosis induced by erastin2 and cyst(e)inase, RSL3 and ML162
SLC1A5	solute carrier family 1 member 5	amino acid transporter feeding glutaminolysis	inhibition or knockdown suppresses ferroptosis induced by amino acid/cystine deprivation
GLS2	glutaminase 2	converts glutamine to glutamate	inhibition or knockdown suppresses ferroptosis induced by erastin or amino acid/cystine deprivation
GOT1	glutamic-oxaloacetic transaminase 1	involved in synthesis of α-ketoglutarate from glutamate	inhibition suppresses ferroptosis induced by erastin or amino acid/cystine deprivation
microRNA 137		targets and regulates SLC1A5 levels	overexpression suppresses SLC1A5 and confers resistance to erastin- and RSL3-induced ferroptosis, while inhibition sensitizes to ferroptosis
G6PD	glucose-6-phosphate dehydrogenase	involved in pentose phosphate pathway	knockdown suppresses erastin-induced ferroptosis in non-small cell lung cancer cells Calu-1
			knockdown suppresses ferroptosis induced by amino acid/cystine deprivation
PGD	phosphoglycerate dehydrogenase	involved in pentose phosphate pathway	knockdown suppresses erastin-induced ferroptosis in non-small cell lung cancer cells Calu-1

그림 3 . Ferroptosis 조절 개요

 

(A-D) 철 대사 와 트랜스페린, TFRC 및 페리티노파지를 포함한 조절 자는 세포 내 불안정 철 풀(LIP) 수준을 증가시켜 지질 과산화 및 페로프토시스를 조절 합니다.

 

(E) 글루타민과 글루타민 분해는 지방산 합성을 위한 구연산염과 같은 전구체를 공급하여 페로프토시스를 촉진 합니다.

 

(F) Mevalonate 경로의 대사산물인 isopentenyl pyrophosphate(IPP)는 콜레스테롤 생합성, isopentenylation, Sec-tRNA 및 CoQ10 생산에 중요하며 FIN56에 의해 유도되는 ferroptosis를 조절합니다.

 

(G) Transsulfuration pathway 는 glutathione 합성을 위해 cystathionine 에서 cysteine을 공급함으로써 ferroptosis에 대한 감수성에 영향을 미친다 .

 

(H) GPX4 는 지질 과산화수소를 해독하여 페로프토시스의 주요 음성 조절인자입니다 . 

GPX4의 활성에는 GSH와 셀레늄이 필요합니다. 

아실-CoA 합성효소 장쇄 패밀리 구성원 4(ACSL4) 및 리소포스파티딜콜린 아실트랜스퍼라제 3(LPCAT3) 의 활성에 의해 조절되는 PL 에서 에스테르화된 아라키도노일(AA)/아드레노일(AdA)의 양이 페로프토시스 에 대한 민감성을 정의합니다. 

PUFA의 직접적인 산소화 에 의한 LOX 는 독성 PL 하이드로퍼옥사이드를 생성 합니다.

 

TF, transferrin; TFRC, transferrin receptor; STEAP3, six-transmembrane epithelial antigen of prostate 3; NCOA4, nuclear receptor coactivator 4; GLS2, glutaminase 2; GOT1, glutamic-oxaloacetic transaminase 1; FPP, farnesyl pyrophosphate; DMT1, divalent metal transporter 1; SQS, squalene synthase; Met, methionine; Cys, cysteine.

 

Table 3. Regulation of Ferroptosis by Cancer Metabolism

 

GeneProteinFunctionModulatory EffectReferences

TP53	wild-type p53	tumor suppressor	knockout/knockdown increases cystine uptake and suppresses ferroptosis in osteosarcoma U2OS and breast cancer MCF7 cells
			knockdown sensitizes colorectal cancer HCT116 and SW48 cell to ferroptosis
			stabilization with nutlin-3 in mouse embryonic fibroblasts, HT-1080 fibrosarcoma, renal cancer Caki-1, and osteosarcoma U2OS cells suppresses ferroptosis
	mutated p533KR	triple acetylation-defective mutant (K117/161/162) that fails to induce cell-cycle arrest, senescence, and apoptosis	retains the ability to regulate SLC7A11 expression and induce ferroptosis
	Mutated p53S47	nonsynonymous single-nucleotide polymorphism at codon 47 in African-descent populations	impaired ferroptosis induction in p53S47 knockin MEF cells
			intact ferroptosis induction in E1A/Ras-transformed p53S47 knockin cells
	mutated p534KR	quadruple acetylation-defective mutant (K98/117/161/162) that fails to induce cell-cycle arrest, senescence, and apoptosis	impaired ferroptosis induction and loss of tumor-suppressor activity
	mutated p53	missense mutations (R273H, R175H) that impair p53 sequence-specific binding to DNA	accumulated mutant-p53 protein sensitizes esophageal and lung cancer cells to ferroptosis
BAP1	BRCA1-associated protein 1	nuclear deubiquitinating epigenetically regulates gene expression	promotes ferroptosis by repression of SLC7A11 expression
CDKN1A	cyclin-dependent kinase (CDK) inhibitor 1A (p21)	inhibits CDK causing cell-cycle arrest	overexpression confers resistance to erastin2-induced ferroptosis (by retarding GSH depletion?)
SAT1	spermidine/spermine N1-acetyltransferase 1	acetylates spermidine and spermine	transcription target of p53 overexpression leads to ferroptosis upon ROS stress knockout suppresses p53-induced ferroptosis
SOCS1	suppressor of cytokine signaling 1	cytokine-induced negative regulators of cytokine signaling	regulates p53 expression and sensitizes cells to ferroptosis
TP63	ΔN tumor protein 63α	oncogene	orchestrates GSH metabolism overexpression confers resistance to erastin- or RSL3-induced ferroptosis, while knockdown enhances ferroptosis
OTUB1	ovarian tumor (OTU) family member deubiquitinase	directly interacts with SLC7A11 and regulates SLC7A11 stability	suppresses ferroptosis by stabilizing SLC7A11 knockout sensitizes to erastin-induced ferroptosis

*********************************************************