Dissecting the intrinsic and extrinsic regulators of prostate cancer dormancy in the bonemicroenvironment.

剖析骨微环境中前列腺癌休眠的内在和外在调节因子。

基本信息

批准号：
10743406
负责人：
Mostafa Nasr
金额：
$ 4.12万
依托单位：
UNIVERSITY OF SOUTH FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-11 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10743406
关键词：
ACVR1 gene Ablation Address Aging Agreement American Androgens Apoptosis Apoptotic Arteries BCL-2 Protein BCL1 Oncogene BCL2L11 gene BMP7 gene Benign Bioinformatics Biological Bioluminescence CASP3 gene CDK4 gene Cancer Etiology Cancer Patient Castration Cell Communication Cell Cycle Progression Cell Cycle Regulation Cell Line Cells Cessation of life ChIP-seq Coculture Techniques Cytometry DNA Binding Data Diagnosis Disease model Ectopic Expression Fluorescence-Activated Cell Sorting Gene Expression Profiling Genes Genetic Growth Hemostatic Agents Histologic Homeostasis Human Immunocompetent Impairment In Vitro Knowledge LAPC4 Lesion Limb structure Literature Malignant Neoplasms Malignant neoplasm of prostate Metastatic Neoplasm to the Bone Metastatic Prostate Cancer Methods Modeling Molecular Molecular Target Mus Neoplasm Metastasis Operative Surgical Procedures Parents Pathway Analysis Pathway interactions Patients Phase Phosphorylation Pre-Clinical Model Primary Neoplasm Protein Region Proteins Publishing Recurrent disease Relapse Research Research Proposals Resistance Role Sampling Signal Transduction Site Skeleton Small Interfering RNA Stromal Cells Survival Rate Techniques Technology Testing Tissues Up-Regulation Validation Work Xenograft Model Zinc Fingers antagonist bone cancer cell experimental study genetic approach high dimensionality improved in vitro Model in vivo in vivo evaluation inhibitor insight knock-down men mortality mutant neoplastic cell novel novel therapeutic intervention p38 Mitogen Activated Protein Kinase patient prognosis pharmacologic prevent pro-apoptotic protein programs prostate cancer cell prostate cancer cell line prostate cancer model receptor receptor expression single-cell RNA sequencing skeletal stem cells transcription factor transcriptome transcriptome sequencing treatment strategy tumor

项目摘要

Project Summary / Abstract………………………………………………………………………………..…. Metastatic prostate cancer (PC) typically manifests in the skeleton. The lesions arise from disseminated tumor cells (DTCs) that often persist in a dormant state for months to decades after initial primary tumor treatment. Despite our knowledge that DTCs give rise to incurable bone metastatic PC, there remains gaps in our understanding as to the molecular mechanisms underpinning entry and reawakening from the dormancy program. Insight into those mechanisms could yield new therapeutic approaches that would prevent the metastatic relapse and ultimately death of almost 34,500 American men each year. To address this, we have developed a novel model of PC dormancy in agreement with published markers in the literature in vitro and optimized a surgical technique to study dormancy in vivo. We next performed transcriptomic sequencing on PC cells growing under normal, dormant, or reawakening conditions. Bioinformatic analysis of transcript expression and transcription factor (TF) activity revealed positive regulatory domain zinc finger region protein 16 (PRDM16) to significantly increase during entry into dormancy and decrease during exit across mouse (RM1) and human sensitive / castrate resistance cell lines (LAPC4, 22Rv1). Further, we validated PRDM16 upregulation in dormant cells on the protein level both in vitro and in vivo. Genetic silencing of PRDM16 led to a significant reduction in the ability of PC cells to enter dormancy concomitant with decreased expression of anti-apoptotic proteins including BCL-2 and increased pro-apoptotic proteins such as BIM & Noxa. Preliminary data shows that PRDM16 expression in PC cells can be induced by BMP-7 and conversely decreased by the BMP signaling antagonist, noggin, factors with known critical roles in regulating bone homeostasis. Based on our findings, we hypothesize that BMP7 induces PRDM16 expression in early skeletal DTCs, which is critical for the initiation of the dormancy program. In the F99 phase, we will identify PRDM16 transcriptome in PC dormancy and confirm BMP signaling control on its expression using genetic approaches and validation of findings in pre-clinical models. In K00 phase, we will study how aging contribute to the reawakening of dormant cancer cells in the bone microenvironment. We will identify biological and molecular determinants of cancer dormancy using Cherry-niche technology and comprehensive fluorescence-activated cell sorting (FACS), high dimensional mass cytometry, and single cell RNASeq experiments. We will then validate our findings in pre-clinical models and patient tissue sections. Findings from this work will advance our understanding of the intrinsic and extrinsic factors regulating the cancer dormancy program in skeletal metastasis in the context of aging, which is relevant to most prostate cancer patients.

项目摘要 /摘要………………………………………………………………………………………………………………。转移性前列腺癌（PC）通常在骨骼中表现出来。病变来自传播肿瘤细胞（DTC）通常在初次原发性原发性肿瘤后数十年持续几个月治疗。尽管我们知道DTC会产生无法治愈的骨转移PC，但仍有我们对所基于的分子机制的理解差距，从休眠计划。洞悉这些机制可能会产生新的治疗方法每年将防止转移性继电器并最终死亡34,500名美国男性。为了解决这个问题，我们开发了一种与已发布的新型PC休眠模型文献中的标志物在体外并优化了一种研究体内休眠的手术技术。我们接下来在正常，休眠或重新唤醒下生长的PC细胞上进行了转录组测序状况。转录本表达和转录因子（TF）活性的生物信息学分析显示阳性调节结构域锌指蛋白16（PRDM16）在进入期间显着增加跨小鼠（RM1）和人类敏感 / castrate抗性的出口期间休眠和减少细胞系（LAPC4，22RV1）。此外，我们验证了蛋白质上休眠细胞中的PRDM16上调在体外和体内都水平。 PRDM16的遗传沉默导致能力显着降低 PC细胞与抗凋亡蛋白的表达降低的休眠伴随包括Bcl-2和增加的凋亡蛋白，例如BIM和NOXA。初步数据显示 BMP-7可以诱导PC细胞中的PRDM16表达，而BMP则降低信号拮抗剂Noggin，在调节骨稳态中具有关键作用的因素。基于根据我们的发现，我们假设BMP7会影响早期骨骼DTC中的PRDM16表达，这对于休眠计划的倡议至关重要。在F99阶段，我们将确定PC休眠中的PRDM16转录组并确认BMP 使用遗传方法和临床前的发现验证其表达的信号传导控制其表达型号。在K00阶段，我们将研究衰老如何促进休眠癌细胞的重新训练在骨微环境中。我们将确定生物学和分子确定剂使用樱桃元素技术和全面的荧光激活细胞分类的休眠状态（FACS），高维质量细胞仪和单细胞RNASEQ实验。然后我们将验证我们在临床前模型和患者组织切片中的发现。这项工作的发现将推动我们的了解调查骨骼癌症休眠计划的内在和外在因素在衰老的情况下，转移与大多数前列腺癌患者有关。