Biomarker Identification to Increase Adjunctive Cryoablation and Checkpoint-Inhibitor Immunotherapy Response

1检查点抑制剂（CPI）在许多肿瘤类型中显着改善了预后，但仍存在一些问题。 2、巨大的改进空间。因此，大量的临床前和临床研究旨在 3评估可能增加CPI反应率的联合治疗方案，并确定 4种生物标志物，表明患者是否会受益于免疫治疗或对免疫治疗产生抗性。 5冷冻消融是一种有前途的治疗策略，可能能够显着增加CPI反应。 6级。单个转移灶的冷冻消融导致坏死细胞死亡，使细胞膜破裂， 7次交替冻融循环。肿瘤抗原沿着炎性细胞内 8个内容。这种明显的炎症反应激活树突状细胞吸收释放的肿瘤抗原 9，反过来，激活适应性免疫系统攻击身体其他部位的肿瘤部位。 10我已经开发了一种双植入肿瘤模型来研究连续冷冻消融的全身益处。 11，并发现当加入CPI治疗时，冷冻消融显著增加了完全反应。 图12是未消融肿瘤相对于单独CPI的比率。由于对CPI增加冷冻消融的研究很少， 13仍然是重要的工作，以确定生物标志物，以确定哪些肿瘤将受益于冷冻消融术 14添加为了确定反应生物标志物，我得到了我共同发明的独特调查工具的帮助， 15颗粒酶B PET显像。定位于主动免疫介导的肿瘤杀伤位点，颗粒酶B PET 图16允许我们在肿瘤大小变化之前区分CPI应答者和非应答者。我们可以 17在免疫反应仍然活跃时询问它，促进独特的见解。 18在本提案中，在小鼠肿瘤模型中，我将开发优化的临床前冷冻消融和CPI方案。 19跨多个肿瘤线，反映CPI反应性的范围。在这些模型中，我将使用 20粒酶B PET成像，以帮助确定基因组和非基因组生物标志物的反应，并确定 21对冷冻消融和CPI组合的抵抗机制。同时，我也将领导一个 22项评价冷冻消融术增加转移性尿路上皮中帕博利珠单抗应答能力的临床试验 23例癌。从冷冻消融时获得的患者血液和活检标本中，我建议分析 24这些患者中应答生物标志物的肿瘤和全身免疫微环境。这些数据将 25对于指导未来优化临床方案的开发至关重要， 26以增加多种肿瘤类型的全身免疫力。我将在一个非常好的环境中进行这项研究。 27 MGH的研究环境，由免疫学，免疫肿瘤学和免疫学领域的导师专家指导， 28篇放射学他们的指导将有助于确保这个项目的成功和我的成功 第29章转型为独立研究

1 Checkpoint inhibitors (CPI) have significantly improved outcomes in many tumor types, but there is still 2 immense room for improvement. A tremendous amount of preclinical and clinical research thus is aimed at 3 evaluating combination therapeutic regimens that may increase CPI response rate as well as identifying 4 biomarkers that indicate whether a patient will benefit from or be resistant to immunotherapy. 5 Cryoablation is a promising adjunctive strategy that may be able to significantly increase the CPI response 6 rate. Cryoablation of a single metastatic focus causes necrotic cell death, rupturing cell membranes through 7 alternating freeze and thaw cycles. Tumoral antigens are released along with inflammatory intracellular 8 contents. This markedly inflammatory reaction activates dendritic cells to take up the released tumor antigen 9 which, in turn, activates the adaptive immune system to attack tumor sites elsewhere in the body. 10 I have developed a dual-implanted tumor model to investigate the systemic benefit of adjunctive cryoablation 11 and have found that when added to CPI therapy cryoablation significantly increases the complete response 12 rate of the non-ablated tumor relative to CPI alone. As adding cryoablation to CPI is minimally studied, there 13 remains significant work to identify biomarkers to determine which tumors will benefit most by cryoablation 14 addition. To identify response biomarkers I am helped by a unique investigative tool that I co-invented, 15 granzyme B PET imaging. Localizing to the site of active immune-mediated tumor killing, granzyme B PET 16 allows us to distinguish CPI responders from non-responders prior to changes in tumor size. We can then 17 interrogate the immune-response while it is still active, facilitating unique insights. 18 In this proposal, in a murine tumor model I will develop an optimized preclinical cryoablation and CPI regimen 19 across multiple tumor lines reflective of the range of CPI responsiveness. In these models I will then use 20 granzyme B PET imaging to help identify genomic and non-genomic biomarkers of response and identify 21 mechanisms of resistance to the combination of cryoablation and CPI. I will also simultaneously be leading a 22 clinical trial evaluating the ability of cryoablation to increase pembrolizumab response in metastatic urothelial 23 carcinoma. From patient blood and biopsy specimens obtained at the time of cryoablation I propose to analyze 24 the tumoral and systemic immune microenvironment for response biomarkers in these patients. These data will 25 be essential to guide the development of future optimized clinical protocols evaluating the ability of cryoablation 26 to increase systemic immunity across multiple tumor types. I will conduct this research in the excellent 27 research environment of MGH, guided by mentors expert in the fields of immunology, immuno-oncology, and 28 interventional radiology. Their guidance will help ensure the success of this project and my successful 29 transition to an independent research career.