Transcriptional and Epigenetic Regulation of HSC Generated from iPS and ESC

iPS 和 ESC 生成的 HSC 的转录和表观遗传调控

• 批准号: 7675159
• 负责人: ALAN D FRIEDMAN
• 金额: $ 4.1万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-01 至 2009-08-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7675159
• 关键词: Address; Adult; Autologous; B-Lymphocytes; Blood; Bone Marrow Stem Cell; CD34 gene; Cell Differentiation process; Cell Therapy; Cells; Chromatin; Clinical; Development; Disease; Epigenetic Process; Failure; Fibroblasts; GATA1 gene; Gene Silencing; Generations; Goals; Hematopoietic; Human; Lead; Lymphoid Cell; Malignant - descriptor; Malignant Neoplasms; Marrow; Mediating; Mesenchymal Stem Cells; Methodology; Methods; MicroRNAs; Molecular Genetics; Mus; Neoplasms; Patients; Proto-Oncogenes; Protocols documentation; RUNX1 gene; Regulation; Retroviridae; Role; SV40 T Antigens; Safety; Scientist; Signal Transduction; Somatic Cell; Source; Staging; Stem cells; Syndrome; Time; Variant; c-myc Genes; human embryonic stem cell; induced pluripotent stem cell; notch protein; pluripotency; progenitor; programs; small molecule; stem; stem cell division; vector

DESCRIPTION (provided by applicant): The recent discovery that patient-specific induced pluripotent stem cells (iPS) can be generated from somatic cells using defined factors has provided unprecedented opportunities for generating unlimited sources of autologous, transplantable hematopoietic stem-progenitor cells (HSC). The clinical safety of stem cell-like iPS currently remains in question, however, since current iPS methodologies utilize integrating retroviruses expressing proto-oncogenes (e.g., Myc, Klf4, and SV40 T antigen), and thus produce cells with great potential for malignant transformation. Moreover, the efficient generation of adult, long-term engrafting HSC from pluripotent human embryonic stem cells (hESC) currently remains elusive. In this proposal, we assemble an interactive team of basic and translational scientists to address these major problems. To achieve the goal of safely generating and expanding iPS-HSC for treatment of hematologic disorders, we will focus on key developmental, transcriptional, and epigenetic mechanisms that orchestrate the expansion of de novo HSC from bone marrow (BM) stem cells, hESC, and iPS (derived from patient-specific fibroblasts, mesenchymal stem cells (MSC), CD34+ progenitors, and B-lymphoid cells). We will strive to replace ectopically-expressed defined factors with alternative reprogramming protocols that utilize non- integrating vectors, miRNAs, siRNAs, and chromatin-modifying small molecules. The goal of the following four Collaborative Projects is to generate clinically useful, transplantable iPS-HSC that lack malignant potential. Project I: Generation of HSC from hESC and iPS (PI: Zambidis). This project will: generate engraftable human HSC from iPS-derived hemangioblasts expressing CD143 by manipulating upstream signals (Notch), and regulatory factors (CDX-Hox); generate iPS from adult CD34+ HSC; investigate the role of miRNAs that regulate stem cell renewal. Project II. Specification and Expansion of Definitive HSC (PI: Friedman). This project will: investigate regulation of RUNX1 by upstream signals (Wnt, Notch, Hox) that mediate HSC emergence; use iPS generated from RUNX1(-/-) mice, ESC, and Runx1 variants to delineate mechanisms that mediate Runx1 specification of definitive HSC; investigate whether inhibition of HSC differentiation facilitates iPS-HSC formation/expansion using PU.1-/-, C/EBP1-/-, and GATA1-/- somatic cells. Project III. Mechanistic Role of c-Myc in iPS Generation (PI: Dang). This project will: delineate the role of a Myc-responsive miRNA cluster in induced pluripotency; generate iPS lacking using B cells at various stages of development. Project IV. Epigenetic Determinants of Neoplasia in iPS (PI: Baylin). This project will: examine the key shared aspect of both neoplasia and iPS reprogramming: abnormal epigenetic gene silencing; determine the degree to which this occurs via various iPS protocols. By drawing upon the broad clinical expertise at Johns Hopkins, our aim is to ultimately lead a translational program for safe iPS-HSC therapies, within the overall project time frame.

描述（由申请人提供）：最近发现，使用定义的因子可以从体细胞中生成患者特异性诱导的多能干细胞（IP）（IPS），为生成自体，可移植的造血干扰素细胞的无限来源提供了前所未有的机会。但是，由于目前的IPS方法学利用了表达原始癌基因的逆转录病毒（例如MYC，KLF4和SV40 T抗原），因此类似干细胞样IPS的临床安全性仍然存在，因此，产生具有恶性转化潜力的细胞。此外，目前难以捉摸的是，从多能人类胚胎干细胞（HESC）中雕刻出长期的长期HSC的有效产生。在此提案中，我们组装了一个基本和翻译科学家的互动团队，以解决这些主要问题。 To achieve the goal of safely generating and expanding iPS-HSC for treatment of hematologic disorders, we will focus on key developmental, transcriptional, and epigenetic mechanisms that orchestrate the expansion of de novo HSC from bone marrow (BM) stem cells, hESC, and iPS (derived from patient-specific fibroblasts, mesenchymal stem cells (MSC), CD34+ progenitors, and B淋巴细胞）。我们将努力用异位表达的定义因子代替替代的重编程协议，这些方案利用非整合矢量，miRNA，siRNA和染色质修饰小分子。以下四个协作项目的目标是产生缺乏恶性潜力的临床上有用的，可移植的IPS-HSC。 项目I：HESC和IPS（PI：Zambidis）的HSC产生。该项目将：通过操纵上游信号（Notch）和调节因素（CDX-HOX）来产生来自IPS来源的血管细胞的植入人HSC；从成人CD34+ HSC产生IP；研究调节干细胞更新的miRNA的作用。 项目II。确定HSC（PI：Friedman）的规范和扩展。该项目将：调查介导HSC出现的上游信号（Wnt，Notch，HOX）对RUNX1的调节；使用Runx1（ - / - ）小鼠，ESC和Runx1变体生成的IP，以描绘介导确定HSC的Runx1规范的机制；研究HSC分化的抑制是否使用PU.1 - / - ，C/EBP1 - / - 和GATA1 - / - 体细胞促进IPS-HSC的形成/扩展。 项目三。 C-MYC在IPS生成中的机械作用（PI：DANG）。该项目将：描述MYC响应性miRNA群集在诱导多能性中的作用；在开发的各个阶段都会生成缺乏使用B细胞的IP。 项目IV。 IPS（PI：Baylin）中肿瘤的表观遗传决定因素。该项目将：检查肿瘤和IPS重编程的关键共同方面：异常表观遗传基因沉默；确定通过各种IPS协议发生的程度。通过借鉴约翰·霍普金斯（Johns Hopkins）的广泛临床专业知识，我们的目标是在整个项目时间范围内领导安全IPS-HSC疗法的转化计划。

项目成果

Running the full human developmental clock in interspecies chimeras using alternative human stem cells with expanded embryonic potential.

• DOI: 10.1038/s41536-021-00135-1
• 发表时间: 2021-05-17
• 期刊: NPJ Regenerative medicine
• 影响因子: 7.2
• 作者: Thomas J;Zimmerlin L;Huo JS;Considine M;Cope L;Zambidis ET
• 通讯作者: Zambidis ET

HMGA1 reprograms somatic cells into pluripotent stem cells by inducing stem cell transcriptional networks.

• DOI: 10.1371/journal.pone.0048533
• 发表时间: 2012
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Shah SN;Kerr C;Cope L;Zambidis E;Liu C;Hillion J;Belton A;Huso DL;Resar LM
• 通讯作者: Resar LM

Integrated Genomic Analysis of Diverse Induced Pluripotent Stem Cells from the Progenitor Cell Biology Consortium.

• DOI: 10.1016/j.stemcr.2016.05.006
• 发表时间: 2016-07-12
• 期刊: Stem cell reports
• 影响因子: 5.9
• 作者: Salomonis N;Dexheimer PJ;Omberg L;Schroll R;Bush S;Huo J;Schriml L;Ho Sui S;Keddache M;Mayhew C;Shanmukhappa SK;Wells J;Daily K;Hubler S;Wang Y;Zambidis E;Margolin A;Hide W;Hatzopoulos AK;Malik P;Cancelas JA;Aronow BJ;Lutzko C
• 通讯作者: Lutzko C