The Role of Oncostatin M in the PDAC tumor microenvironment and macroenvironment

项目摘要/摘要 胰腺癌(PDAC)由于治疗难治，5年内死亡率高达90%。 肿瘤的转移性，以及肿瘤引起的频繁、严重的恶病质或无意中的体重减轻。 肿瘤细胞与宿主细胞局部相互作用，以指定高度促促结缔组织生长的微环境，但也诱导 宏观环境的系统性变化，包括脂肪和肌肉的浪费。炎性细胞因子，如 作为IL-6/gp130家族的成员，包括IL-6和LIF，对微环境有贡献 以及PDAC中的宏观环境。IL-6和LIF促进肿瘤炎症和进展，两者都 与恶病质有关。然而，对另一个IL-6细胞因子家族，包括肿瘤抑素M，人们知之甚少 (OSM)。初步研究支持OSM在PDAC中发挥核心作用。表达了OSM及其受体OSMR 在人类和小鼠肿瘤中。OSMR在肿瘤中的高表达与PDAC患者的生存不良有关。 PDAC瘤小鼠血浆中OSM水平升高。OSM在体外增加人OSMR的表达 小鼠肿瘤细胞和基质细胞表现出前馈调节。OSM诱导的压实 肿瘤细胞/成纤维细胞共培养中的肿瘤球体。相反，从主机中删除OSM，但不删除OSMR 减少肿瘤微环境中的胶原沉积。虽然骨骼肌萎缩，但没有受到影响， 从宿主中删除OSM/OSMR会加速心脏损耗和心功能障碍，并总体上减少 对PDAC肿瘤小鼠的活动性。这些数据表明，OSM/OSMR信号调节肿瘤 PDAC的微观环境和宏观环境。在这里，我们将在两个目标上检验以下假设：1) 肿瘤细胞上的OSM信号通过成纤维细胞促进了PDAC的促结缔组织生长表型。2)OSM 心脏中的信号对PDAC恶病质患者的心脏质量和功能具有保护作用。对于AIM 1，OSM信令 将在使用人类和小鼠肿瘤和成纤维细胞的单一培养和联合培养研究中进行操纵； OSM或OSMR基因缺失的小鼠将被原位移植胰腺癌细胞；肿瘤细胞被删除 因为OSMR将被植入野生型小鼠体内；所有这些都将通过细胞和分子分析进行评估。对于AIM 2， OSM和OSMR基因敲除小鼠将被植入PDAC肿瘤细胞，并评估体重变化， 身体成分、力量/功能、心脏功能和器官损耗。肌肉和心脏将被收获 组织形态计量学和分子分析。培训计划将为申请者提供CRISPR方面的技能， 单细胞测序、组学数据解释、组织免疫表型、癌症模型、心脏 功能障碍和口头和书面交流。培训将在印第安纳大学学院进行 医学，一个高度合作和资源丰富的机构，在特蕾莎·齐默斯博士的直接建议下， 著名癌症恶病质研究人员。简而言之，实习生将成长为一名独立调查员 在培训环境中。

Project Summary/Abstract Pancreatic adenocarcinoma (PDAC) carries a 90% 5-year mortality due to the treatment refractory, highly metastatic nature of the tumor and frequent, severe cachexia or unintentional weight loss induced by tumor. Tumor cells interact with host cells locally to specify a highly desmoplastic microenvironment, but also induce systemic changes in the macroenvironment, including wasting of fat and muscle. Inflammatory cytokines, such as members of the Interleukin-6/GP130 family including IL-6 and LIF, contribute to both the microenvironment and macroenvironment in PDAC. IL-6 and LIF promote tumor inflammation and progression, and both have been implicated in cachexia. However, less is known about the other IL-6 family of cytokines, including Oncostatin M (OSM). Preliminary studies support a central role for OSM in PDAC. OSM and its receptor, OSMR, are expressed in human and murine tumors. High tumor expression of OSMR correlates with poor survival for PDAC patients. OSM was elevated in plasma of mice with PDAC tumors. In vitro, OSM increased expression of OSMR in human and murine tumor cells and stromal cells demonstrating feed-forward regulation. OSM induced compaction of tumor spheroids in tumor cell/fibroblast co-cultures. In contrast, deletion of OSM but not OSMR, from hosts decreased collagen deposition in the tumor microenvironment. While skeletal muscle wasting, was unaffected, deletion of OSM/OSMR from the host accelerated cardiac wasting and cardiac dysfunction and reduced overall activity in mice with PDAC tumors. These data demonstrate that OSM/OSMR signaling modulates both the tumor microenvironment and macroenvironment in PDAC. Here we will test the following hypotheses in two Aims: 1) OSM signaling on tumor cells through fibroblasts promotes the desmoplastic phenotype in PDAC. 2) OSM signaling in the heart is protective of cardiac mass and function in PDAC cachexia. For AIM 1, Osm signaling will be manipulated in monoculture and co-cultures studies using human and murine tumor and fibroblast cells; mice deleted for OSM or OSMR will be orthotopically implanted with pancreatic cancer cells; tumor cells deleted for OSMR will be implanted in wildtype mice; all will be evaluated by cell and molecular analysis. For AIM 2, OSM and OSMR knockout mice will be implanted with PDAC tumor cells and evaluated for body weight change, body composition, strength/function, cardiac function, and organ wasting. Muscle and heart will be harvested for histomorphometry and molecular analysis. The training plan will provide the applicant with skills in CRISPR, single cell sequencing, interpretation of omics data, immunophenotyping of tissues, cancer modeling, cardiac dysfunction and oral and written communication. Training will take place at Indiana University School of Medicine, a highly collaborative and resourced institution, under the direct advisement of Dr. Teresa Zimmers, a well-published cancer cachexia researcher. Summarily, the trainee will grow as an independent investigator within the training environment.