Clonal drivers of resistance to immune checkpoint blockade in liver malignancies

肝脏恶性肿瘤抵抗免疫检查点阻断的克隆驱动因素

• 批准号: 10549797
• 负责人: Giannicola Genovese
• 金额: $ 18.56万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-12 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10549797
• 关键词: Address; Advanced Malignant Neoplasm; Architecture; Automobile Driving; Bar Codes; Behavior; Biological; CRISPR/Cas technology; Cancer Biology; Cancer Etiology; Cancer Model; Cancer Patient; Cells; Cessation of life; Clinical; Clonal Evolution; Clonal Expansion; Clustered Regularly Interspaced Short Palindromic Repeats; Cytotoxic Chemotherapy; Development; Disease; Disease Progression; Ecosystem; Epigenetic Process; Epithelium; Event; Experimental Models; Fishes; Fluorescence; Funding; Generations; Genetic; Genetically Engineered Mouse; Genomics; Hepatobiliary; Heterogeneity; Human; Human Characteristics; Immune; Immune Evasion; Immune response; Immunocompetent; Immunomodulators; Immunotherapy; Kidney; Knowledge; Libraries; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of liver; Metabolic; Methods; Mission; Molecular; Mus; Nature; Nivolumab; Oncologist; Outcome; Pathologic; Patients; Phase; Population; Recovery; Recurrence; Recurrent disease; Reporter; Research; Research Design; Resistance; Sampling; Signal Transduction; Sorting; Suicide; Technology; Therapeutic; Transplantation; Tumor-Derived; United States National Institutes of Health; Validation; adaptive immune response; cancer cell; cancer therapy; cancer type; checkpoint modulation; chemotherapy; design; experience; human disease; immune checkpoint; immune checkpoint blockade; liver cancer model; mortality; mosaic; multiple omics; new technology; novel; patient subsets; pressure; response; targeted treatment; therapy resistant; tool; tumor; weapons

PROJECT SUMMARY Background and relevance to NIH mission. Immune checkpoint blockade based therapy has quickly become the treatment paradigm for several advanced cancer including liver malignancies providing the clinical oncologist with a formidable weapon to achieve dramatic and durable tumor response. The widespread clinical use of modulators of the immune checkpoint, however, has clearly shown that a subset of patients are intrinsically poorly responsive to such treatments, while others might develop recurrent disease after an initial response. While this phenomenon is widely recognized the mechanisms underlying intrinsic and acquired resistance to immune therapy are still poorly understood, posing an urgent need for the development of novel technological tools to study and predict which clones within a tumor will likely drive recurrence. Research design. To investigate the cancer cell intrinsic mechanisms of adaptation and the tumor clonal dynamics in response to immune-checkpoint blockade in an autochthonous experimental model of cancer we will barcode somatic mosaic GEM models of liver cancer to look into the clonal drivers of resistance to immune- checkpoint blockade leveraging a CRISPR-Cas9 based clonal recovery method. Methods. We have generated somatic mosaic cancer models which faithfully recapitulate the biological behavior, the genomic complexity and functional heterogeneity of human disease. To investigate the clonal response to immune therapy malignant cells will be transduced with a dual reporter/suicide cassette barcoded lentiviral library that enables the precise recovery and expansion of any given barcoded clone by using a CRISPR/Cas9 based “fishing” method. These novel technology will be instrumental in the isolation and characterization of those clones/malignant cell populations that are intrinsically prone to evade the immune response or that stochastically acquire the ability to evade the adaptive immune response in the context of immune competent models of cancer. Genetic, molecular and metabolic characterization of such populations will shed light on the mechanisms driving the evasion from immune checkpoint blockade. Ultimately, we will gain fundamental information about clonal dynamics during disease progression in renal malignancies, in addition, by enabling the recovery of single clones and the generation of clonal avatars, this approach would help understanding the relative contribution of intrinsic cell plasticity vs. stochastic genetic events in driving disease recurrence. The technology would eventually demonstrate broader applications in the field of cancer biology and provide novel knowledge and valuable research tools to tackle different cancer types, particularly those characterized by a poor response to immune checkpoints modulation.

项目摘要 背景和与NIH使命的相关性。基于免疫检查点阻断的治疗已迅速成为 包括肝脏恶性肿瘤在内的几种晚期癌症的治疗模式，为临床肿瘤学家提供了 一个强大的武器来实现戏剧性和持久的肿瘤反应。广泛的临床应用 然而，免疫检查点的调节剂已经清楚地表明，一部分患者本质上是 对此类治疗反应不佳，而其他人在最初反应后可能会出现疾病复发。 虽然这一现象被广泛认识到，但内在和获得性耐药性的机制仍有待进一步研究。 免疫治疗仍然知之甚少，迫切需要开发新的技术， 研究和预测肿瘤内哪些克隆可能导致复发的工具。 研究设计。探讨肿瘤细胞的内在适应机制和肿瘤克隆形成的机制。 在一个本地的癌症实验模型中， 将对肝癌的体细胞嵌合GEM模型进行条形码化，以研究对免疫- 检查点阻断利用基于CRISPR-Cas9的克隆恢复方法。方法.我们已经生成 体细胞嵌合体癌症模型，忠实地概括了生物学行为，基因组复杂性， 人类疾病的功能异质性。探讨恶性肿瘤免疫治疗的克隆反应 细胞将用双报告基因/自杀盒条形码化的慢病毒文库转导， 通过使用基于CRISPR/Cas9的“钓取”方法回收和扩增任何给定的条形码化克隆。这些 新技术将有助于分离和表征这些克隆/恶性细胞 那些天生倾向于逃避免疫反应的人群，或者那些随机获得免疫反应能力的人群， 在癌症的免疫活性模型的背景下逃避适应性免疫应答。遗传的，分子的 这些人群的代谢特征将揭示驱动逃避的机制， 免疫检查点阻断。最终，我们将获得有关克隆动态的基本信息， 此外，通过使单个克隆的恢复和 克隆化身的产生，这种方法将有助于理解内在细胞的相对贡献 可塑性与随机遗传事件驱动疾病复发。这项技术最终 展示了在癌症生物学领域更广泛的应用，并提供了新的知识和有价值的 研究工具，以应对不同类型的癌症，特别是那些以免疫反应差为特征的癌症。 检查点调制