Mechanisms for GH action on epithelial cells

GH 对上皮细胞的作用机制

基本信息

批准号：
10578571
负责人：
SHLOMO MELMED
金额：
$ 36.74万
依托单位：
CEDARS-SINAI MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-04-01 至 2028-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10578571
关键词：
3-Dimensional Affect Age Aging Apoptosis Attenuated CRISPR/Cas technology Cell Aging Cell Proliferation Cells Chromosomal Instability Coculture Techniques Colon DNA DNA Damage DNA Repair DNA Repair Pathway DNA-PKcs Data Devices Endocrine Epithelial Cells Epithelium Fibroblasts Gene Expression Genes Genomics Growth Growth Hormone Receptor Homeostasis Human IGF1 gene In Vitro Individual Intestines JAK2 gene Knockout Mice Left Lentivirus Malignant Neoplasms Mediating Metabolic Metabolic dysfunction Metformin Microfluidic Microchips Modeling Mus Mutation Neoplasms Nonhomologous DNA End Joining Oncogenes Organoids Pathway interactions Patients Peripheral Phenotype Phosphoric Monoester Hydrolases Phosphorylation Inhibition Pluripotent Stem Cells Proliferating Publishing Rattus Receptor Signaling Regulation Role Signal Transduction Somatotropin Somatropin Specimen TP53 gene Telomere Shortening Testing Tissue-Specific Gene Expression Tissues Transgenic Mice Transgenic Model Ultraviolet Rays aged antagonist autocrine chemotherapy cohort colon growth design ds-DNA experimental study genomic profiles homologous recombination hormonal signals hormone regulation in vivo induced pluripotent stem cell inhibitor innovation mouse model novel overexpression paracrine pegvisomant pharmacologic receptor repaired response senescence single-cell RNA sequencing stressor tissue degeneration transcriptome sequencing

项目摘要

PROJECT SUMMARY Accumulated damaged DNA, if left unrepaired, may disrupt tissue homeostasis, enabling a milieu favoring tissue degeneration, cancer, and metabolic dysfunction. DNA damage response and repair pathways are attenuated with aging and may also result in cell senescence, with senescence-associated secretory phenotype (SASP) promoting DNA damage and cell proliferation in neighboring cells, or in apoptosis. Non-pituitary growth hormone (npGH), synthesized locally in peripheral tissues, is recognized by the widely expressed GH receptor (GHR), and acts through autocrine/paracrine mechanisms. Our preliminary results show that npGH is induced by DNA damage in aged human colon tissues, in induced pluripotent stem cell (iPSC)-derived human colon 3D organoids, in hypophysectomized rat colon lacking endocrine GH, and in senescent colon cells. In turn, npGH suppresses DNA repair, further reinforcing DNA damage and resulting in chromosomal instability. We propose a novel hypothesis supporting the role of local epithelial GH as an adverse determinant of age-associated DNA damage accumulation and as a marker of epithelial aging. We will assess npGH induction and DNA damage in aged colon organoids, and Colon Intestine-Chip microfluidic devices. We will examine npGH expression in human colon tissue specimens derived from age- determined cohorts, assess spatial genomic profiles in conjunction with cells expressing npGH, and elucidate npGH as a cause of chromosomal instability. We will explore signaling mechanisms for npGH regulation of DNA damage response activity, senescence pathways, and DNA damage repair. Epithelial GHR signaling in vitro will be blocked with the GHR antagonist pegvisomant, a JAK2 inhibitor, and metformin. We will assess npGH actions in aged tissues derived from colon-specific and universal GHR knockout mice, and in GH overexpressing transgenic mice, and also assess npGH paracrine effects on colon epithelial somatic copy number alteration (sCNA) and respective genes affected by sCNA. We will study whether npGH is expressed in senescent cells, and whether it affects adjacent cell DNA damage and senescence. We will test paracrine npGH effects on senescence in Colon Intestine-Chip devices co-cultured with human colon fibroblast npGH transfectants, and will confirm in vitro results in transgenic models overexpressing GH or in GHR knockout mice. We largely focus on validated human colon epithelial models, yet experiments in this proposal are designed to elucidate a universal mechanism whereby local npGH reinforces epithelial age-associated DNA damage leading to tissue degeneration and or neoplasia.

项目摘要累积受损的DNA（如果未经修复）可能会破坏组织稳态，从而使环境有利组织变性，癌症和代谢功能障碍。 DNA损伤响应和维修途径是随着衰老而减弱，也可能导致细胞衰老，与衰老相关的分泌表型（SASP）促进邻近细胞或凋亡中的DNA损伤和细胞增殖。非垂体增长激素（NPGH）在周围组织中局部合成的激素被广泛表达的GH受体识别（GHR），并通过自分泌/旁分泌机制起作用。我们的初步结果表明NPGH是诱导的通过诱导多能干细胞（IPSC）衍生的人类结肠3D的DNA损伤类器官，缺乏内分泌GH的降型大鼠结肠和衰老结肠细胞。反过来，NPGH 抑制DNA修复，进一步增强DNA损伤并导致染色体不稳定性。我们建议一个支持局部上皮GH作为年龄相关DNA的不利决定因素的新假设损害积累和上皮衰老的标志物。我们将评估老年结肠机体和结肠肠芯片的NPGH诱导和DNA损伤微流体设备。我们将检查来自年龄的人类结肠组织标本中的NPGH表达确定的队列，评估与表达NPGH的细胞结合的空间基因组谱，并阐明 NPGH是染色体不稳定性的原因。我们将探索NPGH调节DNA的信号传导机制损伤反应活动，衰老途径和DNA损伤修复。体外上皮GHR信号传导将用GHR拮抗剂PEGVISOMANT，JAK2抑制剂和二甲双胍阻止。我们将评估NPGH行动在源自结肠特异性和通用GHR敲除小鼠的老年组织中，在GH中过表达转基因小鼠，还评估NPGH旁分泌对结肠上皮躯体拷贝数改变的影响（SCNA）和受SCNA影响的基因。我们将研究NPGH是否在衰老细胞中表达，以及它是否影响相邻的细胞DNA损伤和衰老。我们将测试旁分泌NPGH对衰老的影响在与人类成纤维细胞NPGH转染剂共培养的结肠肠芯片设备中，并将确认体外导致过表达GH或GHR敲除小鼠的转基因模型。我们在很大程度上专注于经过验证的人类结肠上皮模型，但该提案中的实验却是设计的为了阐明一种通用机制，局部NPGH加强了与年龄相关的DNA损伤导致组织变性和或肿瘤。

项目成果

期刊论文数量（13）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

WIP1 is a novel specific target for growth hormone action.

DOI：
10.1016/j.isci.2023.108117
发表时间：
2023-11-17
期刊：
ISCIENCE
影响因子：
5.8
作者：
Apaydin, Tugce;Zonis, Svetlana;Zhou, Cuiqi;Valencia, Christian Wong;Barrett, Robert;Strous, Ger J.;Mol, Jan A.;Chesnokova, Vera;Melmed, Shlomo
通讯作者：
Melmed, Shlomo

Generation of isogenic and homozygous MEN1 mutant cell lines from patient-derived iPSCs using CRISPR/Cas9.

使用 CRISPR/Cas9 从患者来源的 iPSC 中生成等基因和纯合的 MEN1 突变细胞系。

DOI：
10.1016/j.scr.2023.103124
发表时间：
2023
期刊：
Stem cell research
影响因子：
1.2
作者：
Even-Zohar,Naomi;Metin-Armagan,Derya;Ben-Shlomo,Anat;Sareen,Dhruv;Melmed,Shlomo
通讯作者：
Melmed,Shlomo

Excess growth hormone suppresses DNA damage repair in epithelial cells