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

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类 患者可以在对改变淋巴细胞活化的药剂的反应中经历有意义的肿瘤消退 和/或扩增，例如IL-2、抗-CTLA 4或IL-2与转移离体扩增的 肿瘤浸润淋巴细胞（TIL）。然而，大多数接受这些治疗的患者未能响应或 实现持久效益。在我们的实验室进行并经其他研究者确认的临床前研究 提供了强有力的证据，即抑制TGF-β信号传导可以显著增强 动物模型中的CD 8+细胞毒性T淋巴细胞（CTL）。我们建议扩展这些研究，以确定 用于临床开发与IL-2组合抑制TGF-β信号传导的药剂的最佳方法， IL-2 + TIL、抗CTLA 4或其他相关免疫操作。临床试验的目的是 在本申请中提出的是改善转移性肿瘤患者的肿瘤反应的速率和质量， 黑色素瘤，并降低这种疾病的发病率和死亡率。 在目的1中，我们提出证实CD 8 + TGFRII-DNR细胞（CD 8 + TGFRII-DNR细胞）的改善的抗肿瘤作用。 小鼠中具有显性负性转化生长因子β受体II的转基因的淋巴细胞 并确定是否可以通过添加以下物质来改善CD 8 + TGFRII-DNR细胞的抗肿瘤活性： IL-2、添加抗CTLA 4和/或添加抗CD 137。此外，我们建议进一步表征 TGF-β减弱抗肿瘤淋巴细胞反应的机制，特别是，以确定是否 TGF-β的作用直接作用于CD 8 + CTL或通过诱导Treg（T调节细胞）间接作用于CD 8 + CTL 其抑制CD 8+和CD 4+抗肿瘤淋巴细胞应答），以确定肿瘤驱动的TGF-β 1的作用。 β在Treg诱导中的作用，并确定Treg是否以及如何减弱CD 8 + CTL介导的肿瘤 排斥反应在目标2中，我们提出创建适合使用的携带TGFRII-DNR基因的逆转录病毒载体 在人临床试验中，为了开发用于人黑素瘤衍生的TIL的转导和扩增的方法， 并表征细胞产物。我们还将使用小鼠黑色素瘤模型来确定最佳的 本发明涉及使TGFRII-DNR CTL的抗肿瘤作用最大化所必需的条件，并且将使用本发明的结果。 实验来指导临床试验的设计。在目标3中，我们建议评估非遗传性 本发明涉及抑制TGF-β的（药理学）方法与免疫疗法的组合。在目标4中，我们 我建议进行概念验证临床试验，其中TGF-β抑制与免疫抑制相结合， 疗法我们建议其中一项临床试验将涉及过继转移黑色素瘤TIL， 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.