Project 1: Evolution of transcriptional programs in lung cancer progression

项目1：肺癌进展中转录程序的进化

• 批准号: 9356947
• 负责人: TYLER E. JACKS
• 金额: $ 42.08万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9356947
• 关键词: Ablation; Address; Affect; Biological Response Modifiers; CRISPR/Cas technology; Cancer Biology; Cancer Etiology; Candidate Disease Gene; Cells; Cessation of life; Collaborations; DNA Sequence Alteration; Data; Development; Diphtheria Toxin; Enzymes; Epigenetic Process; Evolution; Gene Activation; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic; Genetic Engineering; Genetic Transcription; Genetically Engineered Mouse; Genomics; Genotype; HMGA2 gene; Heterogeneity; Human; Immune; Immune Evasion; Immune response; Immunologic Markers; Immunosuppression; Individual; Infiltration; Inflammatory; Investigation; KRAS2 gene; Killer Cells; Laboratories; Lung Adenocarcinoma; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Mediator of activation protein; Messenger RNA; Methods; Modeling; Molecular Profiling; Mutation; Neoplasm Metastasis; Non-Small-Cell Lung Carcinoma; Pathway interactions; Pharmacology; Phenotype; Population; Porcupines; Prevention; Process; Property; Regional Cancer; Series; Signal Pathway; Signal Transduction; Somatic Mutation; TP53 gene; Technology; Testing; Time; United States; Validation; WNT Signaling Pathway; base; cancer cell; cancer genomics; cancer stem cell; cancer type; chemotherapy; cytokine; disorder subtype; gene function; genetic technology; genome editing; immunogenic; improved; innovation; interest; lens; mouse model; mutant; neoplastic cell; new therapeutic target; programs; response; response biomarker; single cell analysis; stem; stem-like cell; targeted treatment; tumor; tumor microenvironment; tumor progression

Project Summary – Project 1 Tumor progression is the product genetic mutations that affect gene function and epigenetic alterations that affect patterns of gene expression and cellular states. Together with the effects of the tumor microenvironment, these changes produce the phenotypes that underlie the biology of cancer. From extensive genomic studies performed over the past decade the genetic landscape of cancer progression is beginning to emerge, but much less is know about how tumors evolve epigenetically. Project 1 is focused on the investigation of tumor evolution through the lens of changes in epigenetic states and gene expression programs. It is anticipated that an improved understanding of these aspects of tumor development will yield new strategies to intervene in this process as well as to treat established tumors. Three Specific Aims will be pursued addressing these questions in the context of well-established genetically-engineered mouse models of human lung adenocarcinoma (LUAD) based on mutations of K-ras (“K”) or the combination of K-ras and p53 (“KP”). Of note, K-ras mutations occur in approximately 30% of human LUAD and targeted therapies do not exist for this subtype of the disease. Preliminary data in support of the Project 1 demonstrates that as tumors progress in the KP model they develop subpopulations of cells that have cancer stem-cell like properties and show markers of response to the Wnt signaling pathway and other cells that markers of Wnt-producing cells that express the enzyme Porcupine (Porcn). It is hypothesized that these Porcn+ cells act as niche cells for the cancer stem-like cells. In Aim 1, a combination of genetic marking, molecular profiling and cell ablation studies will be used to characterized and functionally test these putative niche cells for their importance in tumor development. These studies may identify new targets for the treatment of human lung cancer. Using single cell mRNA sequencing (SCmRNAseq), performed in collaboration with the laboratory of Aviv Regev, the Jacks laboratory has profiled individual cancer cells from K and KP tumors. These data reveal extensive intratumoral and intertumoral transcriptional heterogeneity. Aim 2 of Project 1 will complete the characterization of transcriptional heterogeneity as a function of tumor genotype and tumor progression as well as explore the epigenetic mediators of these effects. Finally, the transcriptional programs of cancer cells are expected to be affected by immune infiltration and anti-tumor immune responses, potentially in a heterogeneous fashion across tumors. These questions will be explored in Aim 3 of Project 1 using SCmRNAseq-based profiling and functional studies of “immunogenic and “nonimmunogenic” tumors. The identification and functional validation of pathways that contribute to immune suppression in this model could reveal new targets of therapy for human lung cancer and other cancer types. In all of the Aims of Project 1, a series of innovative genetic technologies (including lineage tracing, cell marking, cell ablation, and CRISPR/Cas9-mediated genome editing and gene activation) will be used to functionally characterization of genes and pathways identified through profiling.

项目概要 – 项目 1 肿瘤进展是影响基因功能和表观遗传改变的基因突变的产物 影响基因表达和细胞状态的模式以及肿瘤微环境的影响， 通过广泛的基因组研究，这些变化产生了癌症生物学基础的表型。 在过去的十年中，癌症进展的遗传图景开始出现，但是 对于肿瘤如何表观遗传学进化知之甚少。项目 1 的重点是肿瘤的研究。 预计通过表观遗传状态和基因表达程序的变化来进行进化。 更好地了解肿瘤发展的这些方面将产生干预这一问题的新策略 解决这些问题将追求三个具体目标。 在完善的人肺腺癌基因工程小鼠模型的背景下 （LUAD）基于 K-ras（“K”）突变或 K-ras 和 p53 的组合（“KP”）值得注意的是 K-ras 突变。 大约 30% 的人类 LUAD 中发生这种情况，并且针对该亚型不存在靶向治疗 支持项目 1 的初步数据表明，随着肿瘤在 KP 模型中的进展。 他们开发出具有癌症干细胞样特性的细胞亚群，并显示出反应标记 Wnt 信号通路和其他细胞，这些细胞是表达该酶的 Wnt 产生细胞的标记 豪猪（Porcn）被捕获，这些 Porcn+ 细胞充当癌症干细胞样细胞的生态位细胞。 目标 1，将结合基因标记、分子分析和细胞消融研究 对这些假定的利基细胞进行表征并对其在肿瘤发展中的重要性进行功能测试。 研究可能会利用单细胞 mRNA 测序确定治疗人类肺癌的新靶点。 (SCmRNAseq)，与 Aviv Regev 实验室合作进行，Jacks 实验室已分析 来自 K 和 KP 肿瘤的单个癌细胞这些数据揭示了广泛的瘤内和瘤间。 项目 1 的目标 2 将完成转录的表征。 异质性作为肿瘤基因型和肿瘤进展的函数，并探索表观遗传 最后，癌细胞的转录程序预计会受到这些影响的影响。 免疫浸润和抗肿瘤免疫反应，可能以异质的方式跨肿瘤。 这些问题将在项目 1 的目标 3 中使用基于 SCmRNAseq 的分析和功能进行探讨 “免疫原性”和“非免疫原性”肿瘤的研究。 该模型中有助于免疫抑制的途径可能会揭示人类治疗的新靶点 肺癌和其他癌症类型 在项目 1 的所有目标中，一系列创新的基因技术。 （包括谱系追踪、细胞标记、细胞消融以及 CRISPR/Cas9 介导的基因组编辑和基因 激活）将用于对通过分析确定的基因和途径进行功能表征。