Focused Ultrasound Regimens that Synergize with Melanoma Immunotherapy

黑色素瘤的免疫疗法(ITX)已导致临床结果的显著改善。然而， 客观应答率仍然很低(10-30%)，因为在许多患者中T细胞未能渗透到转移瘤中。 因此，辅助途径的翻译可以增强T细胞的渗透和/或解除免疫抑制 肿瘤微环境可以极大地扩大表现出持久反应的黑色素瘤患者的数量 去接受免疫治疗。为了这个目标，我们假设黑色素瘤微环境的扰动与 聚焦超声(FUS)，应用于旨在引发热消融(T-FUS)和/或 微泡空化(M-FUS)，可以刺激免疫反应，既有内在的治疗作用，又有 与可翻译的免疫疗法协同作用。事实上，我们的初步研究表明，即使只适用于 一小部分肿瘤体积，T-FUS和M-FUS都与免疫系统合作控制 黑色素瘤。 这项提案由3个具体目标组成，将用来定义固有的和 通过对肿瘤应用不同的FUS能量方案而引发的适应性免疫反应，识别障碍 肿瘤免疫，并确定更有效地将FUS能量方案与 辅助免疫疗法治疗转移性黑色素瘤。特定目标1将首先描述热学和 T-FUS和M-FUS治疗B16F10黑色素瘤过程中产生的机械能沉积。 然后，使用双侧黑色素瘤模型，我们将识别T-FUS和M-FUS“消融部分”，当 联合一线αPD1免疫疗法治疗黑色素瘤，产生最有效的免疫控制 远处的肿瘤。这些消融部分将在提案的其余部分继续进行。具体目标2 将需要使用αPD1确定选定的FUS能量方案对以下离散因素的影响 T细胞在微环境中的激活、扩增和募集所涉及的一系列步骤 经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利或不利的方面。 目标2下的其他研究将评估应用FUS引起的适应性抗性的机制。 在这些选定的能源制度中。这将使我们能够理解补偿的多样性和强度。 对FUS的抗炎反应，这将是优化FUS与 抗肿瘤免疫疗法。最后，具体目标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FU的组合 能量疗法和免疫治疗药物在我们假设的治疗中将产生最有效的控制 播散性肿瘤。我们认为，这里提出的高度系统和定向的方法更有可能 为T细胞浸润性有限的黑色素瘤患者提供成功的临床治疗。

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.