Targeting immune stroma interactions in pancreatic cancer

胰腺导管腺癌的一个特征是间质纤维化，这是相互作用的结果。 肿瘤细胞、免疫抑制细胞和炎性成纤维细胞之间的关系 Cells‘(PSC)。间质是抑制免疫和靶向肿瘤微环境的重要屏障。 (TME)可以作为一种创新手段来改善效应器免疫细胞对PDAC的访问。此翻译 该提案解决了以HSP90为靶点可以调节TME的组成的假设， 通过其对PSC产生细胞因子的影响，调节T细胞的信号转导，提高 抗PD-1免疫治疗的疗效。我们首次证明，Hsp90抑制物(Hsp90i)限制生长 PSC转录水平的细胞因子分泌。此外，我们公布的数据表明，Hsp90i 在体外和体内抑制关键通路和PDAC的生长。最后，我们展示了Hsp90i提高了 PD-1在小鼠PDAC肿瘤模型中的阻断作用这些数据导致了一项由研究者发起的Ib/II期临床试验 XL888(Hsp90i)和pembrolizumab(抗PD-1)目前在我们机构积累。这项试验的临床成本 因此，这项提议将通过独特的相关研究带来附加值。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说 TME上的Hsp90i。使用试验患者血液的试点数据显示，T细胞的功能反应是 保存完好。对试验患者分离的干细胞样CD8+T细胞的RNA序列和MSigDB分析揭示 STAT和NF-κB信号相关基因的调节，影响T效应细胞的生成和功能。 该提案有三个具体目的：1)评估Hsp90i对PSC和炎性成纤维细胞的影响 在PDAC TME中。从PDAC患者获得的原代PSC将用于培养和有机物质研究 Hsp90的药理或基因改变如何影响PSC。配对的试验活组织检查将审问 Hsp90i联合抗PD-1抗体对TME中PSC或成纤维细胞亚群的影响2)学习 Hsp90i如何改变与PDAC肿瘤相关的免疫调节细胞因子。一个由公认的人类和 小鼠PDAC细胞株，以及来自PDAC患者的有机化合物将用于评估Hsp90i的效果 趋化因子和细胞因子的产生作为影响TME细胞组成的手段。至 补充这些临床前研究、细胞因子和趋化因子特征，以及免疫抑制细胞 将在治疗前后在患者血液和临床试验的配对活检样本中进行评估。 3)研究Hsp90i对T细胞表型和功能的调节作用，以提高PD-1阻断的疗效。我们 我将研究Hsp90i是否限制了NF-κB和JAK/STAT信号转导，以此作为增强T细胞对 PD-1封锁。Hsp90i的免疫活性将通过Hsp90i耐药肿瘤与敏感肿瘤的对比来证实，以及 小鼠模型中的T细胞耗竭抗体。抗PD-1抗体+/-Hsp90i对关键生物标志物的影响 在来自动物研究和临床试验样本的肿瘤浸润性淋巴细胞(TIL)和免疫细胞中得到证实。

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-κB 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-κB 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.