Focused Ultrasound Regimens that Synergize with Melanoma Immunotherapy

黑色素瘤的免疫疗法（ITx）已导致临床结局的显著改善。然而，在这方面， 客观反应率仍然很低（10-30%），因为在许多患者中T细胞不能浸润转移灶。 因此，增强T细胞浸润和/或提高免疫抑制性的辅助方法的翻译是可能的。 肿瘤微环境可以极大地扩大黑色素瘤患者的群体， 免疫疗法为了实现这一目标，我们假设黑色素瘤微环境的扰动， 聚焦超声（FUS），应用于旨在引起热消融（T-FUS）的能量方案，和/或 微泡空化（M-FUS）可以刺激免疫应答，其本质上是治疗性的， 与可翻译的免疫疗法协同。事实上，我们的试点研究表明，即使只应用于 肿瘤体积的一小部分，T-FUS和M-FUS都与免疫系统合作， 黑素瘤 该提案由三个具体目标组成，这些目标将有助于确定先天和 通过对肿瘤应用不同的FUS能量方案引起的适应性免疫应答， 肿瘤免疫，并确定治疗方案，更有效地结合联合收割机FUS能量制度， 用于治疗转移性黑素瘤的辅助免疫疗法。具体目标1将首先描述热特性， 在T-FUS和M-FUS应用于B16 F10黑色素瘤期间产生的机械能沉积。 然后，使用双侧黑色素瘤肿瘤模型，我们将确定T-FUS和M-FUS“消融分数”， 结合一线α PD 1免疫疗法治疗黑色素瘤，可产生最有效的免疫控制， 远处肿瘤这些消融分数将在提案的其余部分继续进行。具体目标2 将需要确定所选FUS能量状态与α PD 1对影响离散因素的影响 涉及T细胞活化、扩增和募集到微环境中的顺序步骤 FUS治疗的肿瘤和远处肿瘤。反过来，这将帮助我们定义免疫细胞激活的优化方案 并了解FUS模式对T细胞介导的肿瘤控制有益或有害的方面。 目标2下的其他研究将评估应用FUS引发适应性抗性的机制 在这些选定的能源制度。这将使我们能够了解代偿性的多样性和强度， FUS的抗炎反应，这将是优化FUS与 抗肿瘤免疫治疗。最后，特异性目标3将确定FUS促进抗体的能力 进入肿瘤，并利用来自先前目标的信息来识别和测试所选FUS的组合 能量疗法和免疫抑制药物的治疗，我们假设将产生最有效的控制 扩散性肿瘤我们认为，这里提出的高度系统化和有针对性的方法更有可能 从而为具有有限T细胞浸润的黑色素瘤患者带来成功的临床治疗。

Immunotherapies (ITx) for melanoma have led to marked improvements in clinical outcomes. However, objective response rates remain quite low (10-30%) because T-cells fail to infiltrate metastases in many patients. Thus, the translation of adjunct approaches that enhance T-cell infiltration and/or lift the immunosuppressive tumor microenvironment could vastly expand the population of melanoma patients exhibiting durable responses to immunotherapy. Toward this goal, we hypothesize that perturbation of the melanoma microenvironment with focused ultrasound (FUS), applied in energy regimens designed to elicit thermal ablation (T-FUS) and/or microbubble cavitation (M-FUS), can stimulate immunologic responses that are both intrinsically therapeutic and synergistic with translatable immunotherapies. Indeed, our pilot studies indicate that, even when applied to only a small fraction of tumor volume, both T-FUS and M-FUS cooperate with the immune system to control melanoma. This proposal is comprised of 3 specific aims that will serve to define differences in the innate and adaptive immune responses that are elicited by applying different FUS energy regimes to tumors, identify barriers to tumor immunity, and ascertain treatment protocols that more effectively combine FUS energy regimes with adjunct immunotherapies for treating metastatic melanoma. Specific Aim 1 will first characterize thermal and mechanical energy deposition generated during the application of T-FUS and M-FUS to B16F10 melanoma. Then, using bilateral melanoma tumor models, we will identify T-FUS and M-FUS “ablation fractions” that, when combined with first line αPD1 immunotherapy for melanoma, yield most efficacious immunological control of a distant tumor. These ablation fractions will be carried forward through the rest of the proposal. Specific Aim 2 will entail determining the impact of the selected FUS energy regimes with αPD1 on discrete factors that influence the sequential steps involved in the activation, expansion, and recruitment of T cells to the microenvironments of FUS-treated and distant tumors. In turn, this will help us define optimized protocols for immune cell activation and understand aspects of FUS modalities that are beneficial or detrimental to T cell-mediated tumor control. Additional studies under Aim 2 will assess the mechanisms of adaptive resistance elicited by application of FUS in these selected energy regimes. This will allow us to understand the diversity and intensity of compensatory anti-inflammatory responses to FUS, which will be a critical aspect in optimizing the combination of FUS with anti-tumor immunotherapy. Finally, Specific Aim 3 will both ascertain the ability of FUS to promote antibody entrance to tumor and utilize information from previous aims to identify and test combinations of selected FUS energy regimens and immunotherapeutic drugs in treatments that we hypothesize will yield most effective control of disseminated tumors. We believe the highly systematic and directed approach proposed here is more likely to lead to successful clinical therapies for melanoma patients with limited T cell infiltration.