Targeting immune stroma interactions in pancreatic cancer

胰腺导管腺癌（PDAC）的一个特征是间质纤维化，这是相互作用的结果 肿瘤细胞、免疫抑制细胞和炎症成纤维细胞之间的关系， 细胞（PSC）。间质是抑制免疫和靶向肿瘤微环境的重要屏障 (TME)可以是改善效应免疫细胞接近PDAC的创新手段。这种平移 该提案提出了靶向Hsp 90可以调节TME组成的假设， 通过其对PSC产生细胞因子的影响，并调节T细胞中的信号转导， 抗PD-1免疫疗法的疗效。我们首次证明，Hsp 90抑制剂（Hsp 90 i）限制生长 以及PSC在转录水平上的细胞因子分泌。此外，我们发表的数据表明Hsp 90 i 在体外和体内抑制关键通路和PDAC生长。最后，我们发现Hsp 90 i提高了 鼠PDAC肿瘤模型中的PD-1阻断。这些数据导致了一项由药物启动的Ib/II期临床试验 XL 888（Hsp 90 i）和派姆单抗（抗PD-1）的研究。这项试验的临床费用 因此，该提案将通过独特的相关研究提供附加值。事实上，这部小说 试验将提供成对的活检和血液，以验证临床前模型中关于影响的假设 热休克蛋白90 i的表达。使用试验患者血液的初步数据显示，T细胞的功能反应是 保存。对来自试验患者的分选的干细胞样CD 8 + T细胞的RNA seq和MSigDB分析揭示了 调节与STAT和NF-κ B B信号传导相关的基因，其影响T效应细胞的产生和功能。 本研究的目的有三：1）探讨热休克蛋白90 i对PSC和炎症成纤维细胞的影响 在PDAC TME中。从PDAC患者获得的原代PSC将用于培养和类器官中以进行研究 Hsp 90药理学或遗传学改变如何影响PSC。试验中的成对活检将询问 Hsp 90 i与抗PD-1 Ab组合对TME中PSC或成纤维细胞亚群的影响。2)研究 Hsp 90 i如何改变与PDAC肿瘤相关的免疫调节细胞因子。一个建立了人类和 鼠PDAC细胞系，沿着来自PDAC患者的类器官将用于评价Hsp 90 i的作用 对趋化因子和细胞因子产生的影响，作为影响TME中细胞组成的手段。到 补充这些临床前研究，细胞因子和趋化因子的签名，连同免疫抑制细胞沿着 将在治疗前和治疗后的患者血液和来自临床试验的配对活检标本中进行评估。 3)确定Hsp 90 i如何调节T细胞表型和功能以增强PD-1阻断的功效。我们 将研究Hsp 90 i是否限制NF-κ B B和Jak/STAT信号传导作为增强T细胞对 PD-1封锁。Hsp 90 i的免疫活性将使用Hsp 90 i抗性肿瘤与敏感肿瘤进行确认，并且 小鼠模型中的T细胞耗竭抗体。抗PD-1 Ab +/-Hsp 90 i对关键生物标志物的影响将在 在来自动物研究和临床试验样品的肿瘤浸润淋巴细胞（TIL）和免疫细胞中证实。

One feature of pancreatic ductal adenocarcinoma (PDAC) is the fibrotic stroma, which is a result of interactions between tumor cells, immune suppressive cells and inflammatory fibroblasts, arising from `pancreatic stellate cells' (PSC). The stroma is a significant barrier restraining immunity and targeting the tumor microenvironment (TME) can be an innovative means to improve access of effector immune cells to PDAC. This translational proposal addresses the hypothesis that targeting Hsp90 can modulate the composition of the TME, through its effects on cytokine production by PSC, and regulate signal transduction in T cells to improve the efficacy of anti-PD-1 immunotherapy. We show for the first time, that Hsp90 inhibitors (Hsp90i) limit growth and cytokine secretion at the transcriptional level from PSC. In addition, our published data indicate Hsp90i inhibits key pathways and PDAC growth in vitro and in vivo. Finally, we show Hsp90i improves the efficacy of PD-1 blockade in murine PDAC tumor models. These data led to an investigator-initiated Phase Ib/II clinical trial of XL888 (Hsp90i) and pembrolizumab (anti-PD-1) now accruing at our institution. The clinical costs of this trial are committed, thus this proposal will deliver added value through unique correlative studies. In fact, this novel trial will provide paired biopsies and blood to validate hypotheses from preclinical models regarding the impact of Hsp90i on the TME. Pilot data using blood of patients on trial show the functional response of T cells is preserved. RNA seq and MSigDB analysis of sorted, stem-like CD8+ T cells from patients on trial reveals modulation of genes related to STAT and NF-B signaling that impact T effector cell generation and function. This proposal has three Specific Aims: 1) To evaluate the impact of Hsp90i on PSC and inflammatory fibroblasts in the PDAC TME. Primary PSC obtained from PDAC patients will be used in culture and in organoids to study how pharmacologic or genetic alteration of Hsp90 influences PSC. Paired biopsies from the trial will interrogate the impact of Hsp90i in combination with anti-PD-1 Ab on subsets of PSC or fibroblasts in the TME. 2) To study how Hsp90i can alter immunomodulatory cytokines relevant to PDAC tumors. A panel of established human and murine PDAC cell lines, along with organoids from PDAC patients will be used to evaluate the effect of Hsp90i on chemokine and cytokine production as a means to influence the cellular composition in the TME. To complement these pre-clinical studies, cytokine and chemokine signatures, along with immune suppressor cells will be assessed in patient blood and paired biopsy specimens from the clinical trial before and after treatment. 3) To determine how Hsp90i modulates T cell phenotype and function to enhance efficacy of PD-1 blockade. We will study if Hsp90i limits NF-B and Jak/STAT signalilng as a mechanism to enhance sensitivity of T cells to PD-1 blockade. The immune activity of Hsp90i will be confirmed using Hsp90i-resistant vs. sensitive tumors, and T cell depleting antibodies in murine models. Effects of anti-PD-1 Ab +/- Hsp90i on key biomarkers will be confirmed in tumor infiltrating lymphocytes (TIL) and immune cells from animal studies and clinical trial samples.