Tumor-TAMs crosstalk enables bypass of oncogenic KRAS dependency in pancreatic cancer

项目摘要/摘要 这项拟议的研究旨在探索延长癌症靶向治疗疾病控制的新策略。 致癌的KRAS(KRAS*)是胰腺癌的关键驱动因素，而药物抑制KRAS的使用 近年来，化学和其他方法得到了广泛的发展。利用多西环素诱导剂 KrasG12D小鼠胰腺p53条件性基因敲除(IKPC)，我们先前的研究表明 70%的PDAC肿瘤在KRAS*耗尽后0.5-1年内复发。我发现肿瘤之间的串扰 细胞和肿瘤相关巨噬细胞(TAMs)促进PDAC对KRAS*耗竭的抵抗。我的 未发表的数据显示，在iKPC细胞中强制表达HDAC5会导致CCL2和CCL7表达上调 和招募TAMS。S100A8+TAMs表达丰富的转化生长因子β使KRAS*非依赖性肿瘤 成长。我推测hdac5-tams-转化生长因子β轴可能是治疗的一般机制。 PDAC中KRAS*耗尽后的耐药性。在这项提案中，我将评估靶向治疗的潜力 通过功能丧失研究几种人和小鼠PDAC模型中的HDAC5、TAMS和转化生长因子β通路。 具体地说，在目标1中，我将确定HDAC5在KRAS*旁路中的重要性，以及对的调节 内源HDAC5的招募。在目标2中，我将利用多维分析来说明表型 KRAS*耗竭后TAMS的动力学和异质性，并论证了TAMS对 肿瘤可以绕过对KRAS的依赖。在目标3中，我将利用Smad4不足的模型来访问本质 KRAS*旁路中转化生长因子β/Smad4通路的激活，阐明转化生长因子β驱动的分子机制 KRAS*旁路。拟议的研究计划将帮助我在一个 学术/医学研究机构。与此同时，我将继续扩大我在生物信息学方面的研究技能， 免疫学、表观遗传学和翻译研究。在我的职业发展方面，我将致力于提高 我在管理实验室，指导博士后和学生，科学写作和演讲，以及寻找 合作，以及其他。安德森癌症中心和罗纳德·德皮尼奥博士的实验室提供了 为我实现这些目标提供了极好的环境。我还成立了一个特别咨询委员会 由朱利奥·德拉塔博士、拉古·卡鲁里博士和阿尼尔班·迈特拉博士组成。他们不仅会为我提供技术支持 支持我提议的学习，但也帮助我寻求教员职位并作为一名独立人士取得成功 调查员。在K22奖的帮助下，我将有一个良好的开端来实现我的长期目标，那就是 作为实验室负责人，继续胰腺癌的基础和翻译研究，包括但不限于有针对性的研究 治疗阻力，并与其他科学家、医生和 制药公司具有团队精神。

Project Summary/Abstract The proposed study is to explore novel strategies to prolong disease control of cancer targeted therapy. Oncogenic KRAS (KRAS*) is the key driver of pancreatic cancer, and pharmaceutical inhibition of KRAS using chemical and other approaches is extensively developed in recent years. Utilizing a doxycycline inducible KRASG12D mouse model with conditional p53 knockout in pancreas (iKPC), our previous publication indicates that 70% PDAC tumors relapsed 0.5-1 year after KRAS* depletion. I identified that the crosstalk between tumor cells and tumor associated macrophages (TAMs) promotes KRAS* depletion resistance in PDAC. My unpublished data shows that forced expression of HDAC5 in iKPC cells leads to upregulation of Ccl2 and Ccl7 and recruitment of TAMs. S100A8+ TAMs express abundant TGFβ that enables KRAS*-independent tumor growth. I hypothesize that the HDAC5-TAMs-TGFβ axis may serve as a general mechanism for therapy resistance after KRAS* depletion in PDAC. In this proposal, I will evaluate the therapeutic potential of targeting HDAC5, TAMs and TGFβ pathway in several human and mouse PDAC models by loss-of-function study. Specifically, In Aim 1, I will determine the essentiality of HDAC5 in KRAS* bypass, and the regulation of TAM recruitment by endogenous HDAC5. In Aim 2, I will utilize multi-dimensional analysis to illustrate the phenotypic dynamics and heterogeneity of TAMs after KRAS* depletion, and demonstrate the essentiality of TAMs for tumors to bypass KRAS* dependency. In Aim 3, I will utilize SMAD4 deficient models to access the essentiality of TGFβ/SMAD4 pathway activation in KRAS* bypass, and delineate the molecular mechanisms of TGFβ-driven KRAS* bypass. The proposed research program will help me to launch an independent faculty position in an academic/medical research institution. Meanwhile, I will keep expanding my research skills in bioinformatics, immunology, epigenetics and translational studies. In terms of my career development, I will devote to improve my skills on managing lab, mentoring postdocs and students, scientific writing and presentation, and seeking for collaborations, among others. MD Anderson Cancer Center and Dr. Ronald DePinho’s laboratory provide an excellent environment for me to achieve these goals. I have also formed an extraordinary advisory committee composed of Drs. Giulio Draetta, Raghu Kalluri, and Anirban Maitra. They will not only provide me technical support for my proposed study, but also assist me to seek a faculty position and succeed as an independent investigator. With the help of K22 award, I will have a good start to achieve my long-term goals, which are to continue basic and translational studies in pancreatic cancer as a lab head, including but not limited to targeted therapy resistance, and to contribute to developing novel cancer therapies with other scientists, physicians and pharmaceutical companies as a team player.