The Role of the Y Chromosome in Bladder Tumor Development, Growth And Progression

Y 染色体在膀胱肿瘤发生、生长和进展中的作用

• 批准号: 10629079
• 负责人: DAN THEODORESCU
• 金额: $ 54.31万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10629079
• 关键词: Address; Aftercare; Aggressive behavior; Alanine; Aminopeptidase; Animal Model; Antibodies; Automobile Driving; B-Lymphocytes; Biological; Bladder; Bladder Neoplasm; CD8-Positive T-Lymphocytes; Cancer Patient; Catalogs; Cell Line; Cell model; Cells; Cessation of life; Chromatin; Chromosomes; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Complement; Data; Data Set; Development; Diagnosis; Diagnostic; Disease; Exposure to; Functional disorder; Gene Deletion; Gene Expression Profile; Gene Expression Profiling; Gene Structure; Genes; Growth; Immune; Immune Evasion; Immune response; Immune system; Immunity; Immunocompetent; Immunophenotyping; Incidence; Knockout Mice; Knowledge; Libraries; Macrophage; Malignant Neoplasms; Malignant neoplasm of urinary bladder; Mediating; Molecular; Mus; Mutagenesis; Myelogenous; Myeloid Cells; Natural Killer Cells; Neoplasm Metastasis; Nitrosamines; Normal Cell; Operative Surgical Procedures; Outcome; Patient-Focused Outcomes; Patients; Phenotype; Precision therapeutics; Process; Prognosis; Proteins; Proteomics; Risk; Role; Smoking; T-Lymphocyte; Technology; Testing; The Cancer Genome Atlas; Therapeutic; Therapeutic Intervention; Tobacco; Translating; Tumor-associated macrophages; Urothelial Cell; Urothelium; Wild Type Mouse; Woman; X Chromosome; Y Chromosome; anti-PD-1; anti-PD1 therapy; autosome; cancer cell; cancer risk; cancer type; chemical carcinogen; chromosome Y loss; combinatorial; differential expression; druggable target; effective intervention; exhaust; functional genomics; genetic signature; histone demethylase; immune cell infiltrate; immune checkpoint blockade; in vivo; inhibitor; intravesical; loss of function; male; men; mortality risk; mosaic analysis; mutant; neoplastic cell; next generation sequencing; novel; overexpression; paralogous gene; patient prognosis; patient stratification; pharmacologic; prognostic; prognostic indicator; programmed cell death protein 1; recombinase; sex; transcriptome sequencing; treatment response; tumor; tumor growth; tumor progression; tumor-immune system interactions; tumorigenesis

PROJECT 2 – SUMMARY Loss of the Y chromosome (LOY) is associated with increased risk of cancer development and death in men with several cancer types. In bladder cancer (BC), a tobacco related malignancy, LOY is seen in ~30% of tumors, yet the biological relevance of this is unknown. Here we address this Knowledge Gap with the Objective to define the molecular mechanisms responsible for LOY driven growth and progression and have Impact by translating these into effective interventions for bladder and other cancer types. Preliminary Data: We examined TCGA BC RNAseq dataset using an LOY gene signature we developed, and found low signature scores associated with reduced patient survival following surgery. To determine if LOY drives BC growth, we developed lineage related male murine cell lines with (Y+) or w/o (Y-) Y chromosome. Y- tumors grew faster than Y+ tumors in immunocompetent mice and cytometric profiling found the former to have increased immunosuppressive tumor-associated macrophages, CD8+ T cells with exhausted phenotype while latter had increased NK cells. In mice (Rag2-/-;Il2rg-/-) that are devoid of T, B and NK cells, Y+ and Y- tumors grew similarly. Transcriptional profiling found four Y chromosome genes differentially expressed in these tumor types, with chromatin organization genes Uty and Kdm5d being the only ones whose expression stratified outcome in TCGA BC patients. Thus, we genetically deleted these genes in Y+ cells and found enhanced tumor growth only in immunocompetent mice while their overexpression in Y- cells reduced their growth. Hence, we test the Hypothesis that Uty and Kdm5d loss enhance BC growth and progression via creation of an immunosuppressive tumor microenvironment in three Specific Aims. In Aim 1 we determine role of Uty and Kdm5d in tumor development and progression using full and urothelial specific conditional Kdm5d or Uty null mice be exposed to the chemical carcinogen BBN. With Project 3 we test the hypothesis that Uty and its X chromosome paralog Utx (aka, Kdm6a) have independent suppressive effects on BC formation and progression by crossing full and urothelial specific conditional Uty and Utx null mice and exposing them to BBN. In Aim 2 we define how Kdm5d and Uty suppress BC and the role of the immune system in this process using defined LOF functional mutants to define the regions on these proteins responsible for tumor growth suppression. With Project 1, we use spatial proteomics to examine immunological infiltrates in relation to mutant and WT Kdm5d and Uty cancer cell expression and various knockout mice or antibody depletion strategy to define which cellular components of the immune system drive tumor growth differences. In Aim 3 we examine ANPEP, the most upregulated gene in Y- tumors that is also a poor prognostic in BC patients and druggable target, as a candidate autosomal effector gene driving Y- tumor growth. We found Y- tumors are more responsive than Y+ tumors to anti-PD-1 and ANPEP depletion in Y- cells made tumors more sensitive to anti-PD-1. Here we will examine the impact of ANPEP deletion or pharmacologic inhibition on the efficacy of anti-PD-1 immune checkpoint blockade therapy.

项目2 -概要 Y染色体丢失（LOY）与男性癌症发展和死亡风险增加有关 几种癌症类型。在膀胱癌（BC）中，烟草相关的恶性肿瘤，LOY见于约30%的肿瘤中， 但其生物学意义尚不清楚。在这里，我们解决这个知识差距的目标， 定义负责LOY驱动生长和进展的分子机制，并通过以下方式产生影响： 将这些转化为膀胱和其他癌症类型的有效干预措施。初步数据：我们检查了 使用我们开发的LOY基因签名的TCGA BC RNAseq数据集，发现签名得分较低 与手术后患者存活率降低相关。为了确定LOY是否推动BC增长，我们开发了 具有（Y+）或w/o（Y-）Y染色体的谱系相关雄性鼠细胞系。Y-肿瘤比Y+肿瘤生长更快 在免疫功能正常的小鼠中，细胞计数分析发现前者具有增加的免疫抑制作用， 肿瘤相关的巨噬细胞，CD 8 + T细胞具有耗尽的表型，而后者具有增加的NK细胞。在 缺乏T、B和NK细胞的小鼠（Rag 2-/-; Il 2 rg-/-），Y+和Y-肿瘤生长相似。转录谱 发现四个Y染色体基因在这些肿瘤类型中差异表达，染色质组织基因 Uty和Kdm 5d是TCGA BC患者中唯一表达分层的结果。因此我们 在Y+细胞中基因删除了这些基因，并发现只有在免疫活性小鼠中肿瘤生长增强 而它们在Y细胞中的过度表达则降低了它们的生长。因此，我们检验了Uty和Kdm 5d 免疫抑制性肿瘤微环境的产生增强了BC的生长和进展 三个具体目标。在目的1中，我们确定Uty和Kdm 5d在肿瘤发生和进展中的作用 使用完全和尿路上皮特异性条件性Kdm 5d或Uty缺失小鼠暴露于化学致癌物BBN。 在项目3中，我们测试了Uty及其X染色体伴侣Utx（aka，Kdm 6a） 通过完全和尿路上皮特异性交叉对BC形成和进展的独立抑制作用 条件性Uty和Utx缺失小鼠并将它们暴露于BBN。在目标2中，我们定义Kdm 5d和Uty如何抑制 BC和免疫系统在此过程中的作用，使用定义的LOF功能突变体来定义区域 这些负责抑制肿瘤生长的蛋白质。在项目1中，我们使用空间蛋白质组学， 检查与突变型和WT Kdm 5d和Uty癌细胞表达相关的免疫浸润， 各种敲除小鼠或抗体耗竭策略，以确定免疫系统的哪些细胞成分 驱动肿瘤生长差异。在目标3中，我们检测了ANPEP，它是Y型肿瘤中上调最多的基因， 也是BC患者的不良预后和药物靶点，作为驱动Y染色体的候选常染色体效应基因， 肿瘤生长我们发现Y-肿瘤比Y+肿瘤对Y-肿瘤中的抗PD-1和ANPEP消耗更敏感。 细胞使肿瘤对抗PD-1更敏感。在这里，我们将研究ANPEP删除或 抗PD-1免疫检查点阻断疗法的功效的药理学抑制。