ENHANCEMENT OF ANTI-TUMOR IMMUNITY BY INHIBITION OF TGF-B SIGNALING IN PATIENS WI

对于转移性黑色素瘤患者的治疗是不够的。转移性黑色素瘤的一个亚型 患者可以在改变淋巴细胞激活的药物作用下进行有意义的肿瘤消退 和/或扩增，例如IL-2、抗CTLA4或IL-2与体外扩增的转移相结合 肿瘤浸润性淋巴细胞(TIL)。然而，接受这些疗法的大多数患者都没有反应或 实现持久效益。在我们的实验室进行并得到其他研究人员确认的临床前研究 提供强有力的证据表明，抑制转化生长因子-β信号转导可以显著增强其抗肿瘤活性。 CD8+细胞毒性T淋巴细胞(CTL)在动物模型中的作用我们建议扩大这些研究，以确定 临床开发联合抑制转化生长因子-β信号转导和IL-2的最佳方法， IL-2+TIL、抗CTLA4或其他相关免疫治疗操作。临床试验的目标是 在这一应用中提出的是提高转移性患者的肿瘤反应率和质量 治疗黑色素瘤，降低这种疾病的发病率和死亡率。 在目标1中，我们建议证实CD8+TGFRII-DNR细胞(CD8+)的抗肿瘤效果 转显性阴性转化生长因子β受体基因的小鼠淋巴细胞 并确定CD8+TGFRII-DNR细胞的抗肿瘤活性是否可以通过加入 IL-2、添加抗CTLA4和/或添加抗CD137。此外，我们建议进一步表征 转化生长因子-β减弱抗肿瘤淋巴细胞反应的机制，特别是为了确定 转化生长因子-β的作用是直接作用于CD8+CTL或通过诱导Treg(T)调节细胞间接发挥作用 它们抑制CD8+和CD4+抗肿瘤淋巴细胞反应)，以确定肿瘤驱动的转化生长因子-1的作用。 β在Treg诱导中的作用以及Treg是否以及如何抑制CD8+CTL介导的肿瘤 拒绝。在目标2中，我们建议创建一种适合使用的携带TGFRII-DNR基因的逆转录病毒载体 在人类临床试验中，为了开发转导和扩增人黑色素瘤来源的TIL的方法， 并对细胞产物进行表征。我们还将使用小鼠黑色素瘤模型来确定最佳 最大限度发挥TGFRII-DNR CTL抗肿瘤作用的必要条件，并将使用 指导临床试验设计的实验。在目标3中，我们建议评估非遗传性 (药理学)结合免疫疗法抑制转化生长因子-β的方法。最后，在目标4中，我们 建议进行概念验证临床试验，其中转化生长因子-β抑制与免疫相结合 心理治疗。我们建议其中一项临床试验将涉及携带TIL的黑色素瘤TIL的过继转移 TGFRII-DNR基因。

Treatment for patients with metastatic melanoma is inadequate. A subset of metastatic melanoma patients can undergo meaningful tumor regression in response to agents that modify lymphocyte activation and/or expansion, for example, IL-2, anti-CTLA4, or IL-2 in combination with transfer of ex vivo expanded tumor-infiltrating lymphocytes (TIL). However, most patients receiving these therapies fail to respond or to achieve lasting benefit. Preclinical studies conducted in our laboratories and confirmed by other investigators provide strong evidence that inhibition of TGF-beta signaling can markedly enhance the anti-tumor activity of CD8+ cytotoxic T-lymphocytes (CTL) in animal models. We propose to extend these studies to determine optimal approaches for clinical development of agents that inhibit TGF-beta signaling in combination with IL-2, IL-2 + TIL, anti-CTLA4, or other related immunotherapeutic manipulations. The goal of the clinical trials proposed in this application is to improve the rate and quality of tumor responses in patients with metastatic melanoma and to reduce the morbidity and mortality from this disease. In Aim 1 we propose to confirm the improved anti-tumor effects of CD8+ TGFRII-DNR cells (CD8+ lymphocytes with the transgene for the dominant negative transforming growth factor beta receptor II) in mouse models and to determine if the anti-tumor activity of CD8+ TGFRII-DNR cells can be improved by addition of IL-2, addition of anti-CTLA4, and/or addition of anti-CD137. Furthermore, we propose to further characterize the mechanisms by which TGF-betaattenuates anti-tumor lymphocyte responses, in particular, to determine if the effects of TGF-beta are directly on CD8+ CTL or indirectly through induction of Treg (T regulatory cells which inhibit CD8+ and CD4+ anti-tumor lymphocyte responses), to determine the role of tumor driven TGF- beta in induction of Treg, and to determine whether and how Treg attenuate CD8+ CTL mediated tumor rejection. In Aim 2, we propose to create a retroviral vector carrying the TGFRII-DNR gene suitable for use in human clinical trials, to develop methods for transduction and expansion of human melanoma-derived TIL, and to characterize the cell product. We will also use mouse melanoma models to determine the optimal conditions necessary to maximize the anti-tumor effects of TGFRII-DNR CTL, and will use the results of the experiments to guide the design of a clinical trial. In Aim 3, we propose to evaluate non-genetic (pharmacologic) approaches to inhibit TGF-beta combined with an immunotherapy. Finally, in Aim 4, we propose to conduct proof of concept clinical trials in which TGF-beta inhibition is combined with an immune therapy. We propose that one of the clinical trials will involve adoptive transfer of melanoma TIL carrying the gene for TGFRII-DNR.