GCDH Addiction in Melanoma

黑色素瘤中的 GCDH 成瘾

• 批准号: 10656574
• 负责人: Steven H Olson
• 金额: $ 72.17万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10656574
• 关键词: Address; Advanced Development; Antibodies; Apoptosis; Apoptotic; Attenuated; Azoles; Brain Neoplasms; Breast Cancer Cell; CASP3 gene; Catabolism; Cell Death; Cell Death Signaling Process; Cell Line; Cell Survival; Cessation of life; Chemicals; Chemoresistance; Clinical; Data; Development; Dissociation; Effectiveness; Enzymes; Evaluation; Foundations; Genetic; Genetic Transcription; Growth; Human; Immune; Immunotherapy; In Vitro; Induction of Apoptosis; Knockout Mice; Liver neoplasms; Lysine; Lysine Degradation Pathway; Malignant Neoplasms; Malignant neoplasm of liver; Malignant neoplasm of prostate; Mammary Neoplasms; Maps; Mediating; Melanoma Cell; Mitochondria; Modality; Modeling; Mole the mammal; Molecular Weight; Monitor; Mus; Neoplasm Metastasis; Outcome; Oxidoreductase; Pathway interactions; Patients; Pharmaceutical Chemistry; Phenocopy; Phenotype; Physiological; Process; Production; Prostatic Neoplasms; Proteins; Regulation; Resistance; Resistance development; Sampling; Signal Pathway; Signal Transduction; Site; Specimen; Testing; Transactivation; Transcription Coactivator; Transcriptional Activation; Tumor Cell Line; Xenograft procedure; activating transcription factor 3; addiction; anti-PD-1; cancer cell; gene function; genome-wide analysis; glutaryl coA; in vivo; inhibitor; liquid chromatography mass spectrometry; malignant breast neoplasm; melanocyte; melanoma; mouse model; new therapeutic target; novel; novel therapeutics; prostate cancer cell; response; small molecule; small molecule inhibitor; targeted treatment; therapy resistant; transcription factor; treatment response; tumor; tumor growth; tumor heterogeneity; ubiquitin-protein ligase

Project Summary In this revised R01 application we propose to characterize a novel mechanism underlying the control of apoptosis by the mitochondrial enzyme GCDH, and to extend the development of small molecule GCDH inhibitors to treat melanoma. In our preliminary results we discovered that addiction to GCDH activity is critical for cell survival in melanoma, but not liver, breast or prostate tumor cells, a discovery that was confirmed in patient specimens where inverse correlation between GCDH expression and survival is seen in melanoma. Our studies reveal induction of apoptosis in melanoma cell lines following GCDH inhibition, a phenotype dependent on the upstream DHTKD1 enzyme. Key in mediating GCDH activities in melanoma is NRF2, which is subjected to glutarylation upon inhibition of GCDH. NRF2 glutarylation promotes its stability and transactivation of an apoptotic UPR signaling consisting of ATF4, ATF3, CHOP and CHAC1, the latter being components of the Unfolded Protein Response (UPR). Inhibition of GCDH effectively induces apoptotic UPR signaling in melanoma but not in liver or breast cancer cells, substantiating the selectivity of the pathway as mapped in cell lines and patient tumor samples. These observations provide the foundation for our hypothesis that GCDH-mediated regulation of NRF2-UPR signaling constitutes a novel pathway controlling melanoma cell survival. Our studies will (i) map the NRF2 glutarylation pathway in melanoma, generate antibodies specific to glutarylated NRF2, monitor NRF2 glutarylation in specimens from melanoma patients both responsive and non-responsive to therapy (ii) assess how GCDH impacts tumor development, progression and response to targeted and immuno therapy in genetic mouse models. YUMM1.7, MaRas, B16F10 lines will be modified to express inducible KD of GCDH before their inoculation into syngeneic WT or GCDH KO mouse models which will be monitored prior and following therapy (iii) advance development GCDH inhibitors as novel therapeutic modalities. SBI-0690564 was confirmed in vitro and in cultured melanoma cells, where it phenocopies genetic inactivation of GCDH and inhibits melanoma tumors in mice. We will characterize and further develop a novel class of inhibitors to target GCDH and determine their effectiveness in culture and in vivo using mouse models. Our studies will establish novel paradigm for GCDH signaling as we define mechanisms underlying GCDH control melanoma tumor fate. Understanding the addiction to GCDH in melanoma provides the foundation for the development and evaluation of novel GCDH inhibitors to selectively target these and possibly other select cancers.

项目概要 在这个修订后的 R01 申请中，我们建议描述一种控制细胞凋亡的新机制 通过线粒体酶GCDH，并扩展小分子GCDH抑制剂的开发来治疗 黑色素瘤。在我们的初步结果中，我们发现对 GCDH 活性的成瘾对于细胞存活至关重要 黑色素瘤，但不是肝脏、乳腺癌或前列腺肿瘤细胞，这一发现已在患者样本中得到证实 其中，黑色素瘤中 GCDH 表达与生存率呈负相关。我们的研究表明 GCDH 抑制后诱导黑色素瘤细胞系凋亡，这是一种依赖于上游的表型 DHTKD1 酶。黑色素瘤中介导 GCDH 活性的关键是 NRF2，它会发生戊二酸化 抑制 GCDH 后。 NRF2 戊二酰化促进其稳定性和凋亡 UPR 的反式激活 信号由 ATF4、ATF3、CHOP 和 CHAC1 组成，后者是未折叠蛋白的组成部分 回应（普遍定期审议）。抑制 GCDH 可有效诱导黑色素瘤中的细胞凋亡 UPR 信号传导，但不能诱导肝脏中的细胞凋亡 或乳腺癌细胞，证实了细胞系和患者肿瘤中绘制的通路的选择性 样品。这些观察结果为我们的假设奠定了基础，即 GCDH 介导的调节 NRF2-UPR 信号通路构成了控制黑色素瘤细胞存活的新途径。我们的研究将 (i) 绘制黑色素瘤中 NRF2 戊二酰化途径的图谱，生成戊二酰化 NRF2 特异性抗体，监测 对治疗有反应和无反应的黑色素瘤患者标本中的 NRF2 戊二酰化 (ii) 评估 GCDH 如何影响肿瘤的发展、进展以及对靶向和免疫治疗的反应 遗传小鼠模型。 YUMM1.7、MaRas、B16F10 系将被修改以表达 GCDH 的诱导型 KD 在接种到同基因 WT 或 GCDH KO 小鼠模型之前，将对其进行监测 后续治疗 (iii) 推进 GCDH 抑制剂的开发作为新的治疗方式。 SBI-0690564 是 在体外和培养的黑色素瘤细胞中得到证实，它复制了 GCDH 的基因失活并抑制 小鼠黑色素瘤。我们将表征并进一步开发一类针对 GCDH 的新型抑制剂 并使用小鼠模型确定其在培养和体内的有效性。我们的研究将建立新颖的 GCDH 信号传导的范例，因为我们定义了 GCDH 控制黑色素瘤肿瘤命运的机制。 了解黑色素瘤对 GCDH 的成瘾性为开发和评估奠定了基础 新型 GCDH 抑制剂可选择性地针对这些癌症以及可能的其他选定癌症。