Dissecting mechanisms of immunotherapy resistance in melanoma patients

剖析黑色素瘤患者免疫治疗耐药的机制

• 批准号: 10231195
• 负责人: Benjamin Izar
• 金额: $ 12.46万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-14 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10231195
• 关键词: Advisory Committees; Angiogenesis Pathway; Biological; Biological Response Modifiers; Biopsy; CRISPR/Cas technology; CTLA4 gene; Cancer Biology; Cancer Patient; Caring; Cell Communication; Cell Line; Cells; Clinical; Clinical Trials; Coculture Techniques; Collaborations; Computer Analysis; Dana-Farber Cancer Institute; Development; Disease; Disease remission; Drug resistance; Enrollment; Environment; Evaluation; Formalin; Genetic; Genetic Screening; Genetic Transcription; Goals; Image; Immune; Immune checkpoint inhibitor; Immunobiology; Immunooncology; Immunotherapy; KDR gene; Knock-out; Ligands; Link; Malignant Neoplasms; Mediating; Melanoma Cell; Mentors; Mentorship; Metastatic Melanoma; Methods; Modeling; Molecular; Molecular Analysis; Nivolumab; PD-1/PD-L1; Paraffin Embedding; Patients; Pharmacology; Phosphotransferases; Physicians; Physiological; Procedures; Protein Analysis; Proteins; Research; Research Personnel; Resistance; Role; Sampling; Scientist; Signal Transduction; Site; Skin Cancer; Specimen; T-Lymphocyte; Technology; Testing; Therapeutic; Tissues; Training; Translating; Tumor-Infiltrating Lymphocytes; Validation; Vascular Endothelial Growth Factors; Work; angiogenesis; anti-PD1 therapy; bevacizumab; biomarker discovery; cancer care; cancer pharmacology; checkpoint therapy; clinical application; clinically relevant; cohort; gain of function; genome-wide; improved; ipilimumab; medical schools; meetings; melanoma; multimodality; multiplexed imaging; novel drug combination; novel therapeutics; pembrolizumab; predicting response; prospective; protein expression; receptor; resistance mechanism; response; single-cell RNA sequencing; spatiotemporal; technological innovation; therapy development; therapy resistant; transcriptome; transcriptome sequencing; treatment strategy; tumor; tumor heterogeneity

Project Summary / Abstract Metastatic melanoma is the most aggressive form of skin cancer and a devastating disease. The development of immune checkpoint inhibitors (ICI) represents a major therapeutic improvement in melanoma care, inducing durable responses in a portion of patients. However, the majority of patients has intrinsic or acquired resistance to ICI and derives no benefit from these therapies. The molecular underpinnings of ICI-resistance are poorly understood. The overarching goal of this proposal is to determine mechanisms of ICI-resistance in melanoma using several experimental and technological innovations. We performed single-cell RNA- sequencing (sc-RNA-seq) and multiplexed imaging of ICI-resistant melanoma tumors, and a genome-scale gain-of-function screen in patient-derived melanoma cell lines and their corresponding tumor infiltrating lymphocytes (TILs). Our sc-RNA-seq analysis revealed a strong association between melanoma cell- autonomous expression of angiogenesis pathways with ICI-resistance. In line with this finding, our functional genetic screen identified KDR (also known as VEGFR2) as top hit of putative mediators of immune escape. KDR is a major receptor for vascular endothelial growth factor (VEGF) and induces the expression of angiogenesis pathways that were identified by sc-RNA-seq. Multiplexed imaging confirmed protein expression of KDR at the invasive tumor front, a key site of tumor-immune interactions in melanoma that predicts response and resistance to ICI. Together, these preliminary results highlight the putative role of KDR and downstream angiogenesis pathway expression in ICI-resistance. This proposal builds on these findings with the specific focus to: 1) dissect the mechanisms of KDR-mediated immune escape, 2) validate transcriptional and protein expression of KDR and its downstream angiogenesis pathways in patients undergoing serial biopsies while receiving ICI therapy, and 3) functionally validate mechanisms of ICI-resistance in isogenic patient-derived cell lines. Results of these studies have the potential to guide novel drug combination strategies that could be rapidly translated into clinical application. Dr. Benjamin Izar is mentored by Dr. Kai Wucherpfennig, a physician-scientist and expert in immuno-oncology, and Dr. Aviv Regev, a pioneer in sc- RNA-seq and computational analyses. Dr. Izar has a committed advisory committee comprised of Drs. Stephen Hodi, Keith Flaherty and Peter Sorger, who will provide additional mentorship and collaboration in immuno-oncology, cancer biology and pharmacology. Dr. Izar has developed a 5-year training plan with a detailed outline of activities that will facilitate his development to an independent investigator. Dr. Izar will leverage an exceptional research environment at Dana-Farber Cancer Institute, Harvard Medical School and the Broad, and a richness of scientific meetings, professional development seminars and didactic coursework.

项目摘要/摘要 转移性黑色素瘤是皮肤癌中最具侵袭性的形式，也是一种毁灭性的疾病。最新进展 免疫检查点抑制剂(ICI)的使用代表了黑色素瘤治疗的重大进步，诱导 部分患者出现持久反应。然而，大多数患者有先天或后天的 对ICI具有抵抗力，并且不能从这些疗法中获益。ICI抗性的分子基础 人们对此了解甚少。这项建议的首要目标是确定ICI的抵抗机制。 利用几项实验和技术创新来治疗黑色素瘤。我们进行了单细胞核糖核酸- 耐ICI黑色素瘤肿瘤的测序(sc-rna-seq)和多重成像，以及基因组规模 患者来源的黑色素瘤细胞系及其相应肿瘤浸润性的功能增强筛选 淋巴细胞(TIL)。我们的sc-RNA-seq分析显示，黑色素瘤细胞- 具有ICI抗性的血管生成途径的自主表达。根据这一发现，我们的功能 基因筛查发现KDR(也称为VEGFR2)是推测的免疫逃逸介质中最受欢迎的。 KDR是血管内皮生长因子(VEGF)的主要受体，并诱导血管内皮细胞生长因子 Sc-RNA-seq鉴定的血管生成途径。多路成像证实蛋白质表达 KDR在侵袭性肿瘤前沿的表达，这是黑色素瘤中肿瘤与免疫相互作用的关键部位，可以预测 对ICI的反应和抵抗。总之，这些初步结果突出了KDR和KDR的假定作用 下游血管生成通路在ICI抵抗中的表达。该提案建立在这些调查结果的基础上 其重点在于：1)剖析KDR介导的免疫逃逸机制；2)验证转录 KDR及其下游血管生成通路在系列血管生成中的蛋白表达 接受ICI治疗时的活检，以及3)从功能上验证ICI耐药的机制。 患者来源的细胞系。这些研究的结果有可能指导新的药物组合 可以迅速转化为临床应用的策略。本杰明·伊扎尔博士由凯博士指导 Wucherpfennig是内科科学家和免疫肿瘤学专家，Aviv Regev博士是sc- Rna-seq和计算分析。伊扎尔博士有一个由伊扎尔博士组成的忠诚的顾问委员会。 Stephen Hodi、Keith Flaherty和Peter Sorger，他们将在 免疫肿瘤学、癌症生物学和药理学。伊扎尔博士制定了一项为期5年的培训计划， 有助于他成长为独立调查员的活动的详细大纲。伊扎尔博士会 利用达纳-法伯癌症研究所、哈佛医学院和 以及丰富的科学会议、专业发展研讨会和教学课程。

项目成果

Alpha-fetoprotein (AFP) as tumor marker in a patient with urothelial cancer with exceptional response to anti-PD-1 therapy and an escape lesion mimic.

• DOI: 10.1186/s40425-018-0394-y
• 发表时间: 2018-09-12
• 期刊: Journal for immunotherapy of cancer
• 影响因子: 10.9
• 作者: Melms JC;Thummalapalli R;Shaw K;Ye H;Tsai L;Bhatt RS;Izar B
• 通讯作者: Izar B

Rapid evolution of acute kidney injury after initial infusion of pembrolizumab in a melanoma patient concurrently treated with RAF/MEK inhibitors.

• DOI: 10.1097/cmr.0000000000000646
• 发表时间: 2020-04
• 期刊: Melanoma research
• 影响因子: 2.2
• 作者: Thummalapalli R;Melms JC;Mier J;Izar B
• 通讯作者: Izar B

Characterization of genetics in patients with mucosal melanoma treated with immune checkpoint blockade.

• DOI: 10.1002/cam4.3789
• 发表时间: 2021-04
• 期刊: Cancer medicine
• 影响因子: 4
• 作者: Buchbinder EI;Weirather JL;Manos M;Quattrochi BJ;Sholl LM;Brennick RC;Bowling P;Bailey N;Magarace L;Ott PA;Haq R;Izar B;Giobbie-Hurder A;Hodi FS
• 通讯作者: Hodi FS