Patient-specific and universal donor blood cell from protein-induced iPS cells

来自蛋白质诱导的 iPS 细胞的患者特异性和通用供体血细胞

• 批准号: 8045716
• 负责人: Kwang-Soo Kim
• 金额: $ 189.64万
• 依托单位: MCLEAN HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-20 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8045716
• 关键词: Address; Asia; Biomedical Research; Blood; Blood Cells; Blood Platelets; Blood typing procedure; Cell Line; Cell Lineage; Cell Therapy; Cells; Clinical; Country; Data; Disease; Embryo; Emergency Situation; Erythrocytes; Fibroblasts; Functional disorder; General Population; Genetic; Human; In Vitro; Life; Location; Low Prevalence; Medical; Medicine; Megakaryocytes; Methods; Patients; Peptides; Preclinical Drug Evaluation; Property; Proteins; Retroviridae; Somatic Cell; Source; Stem cells; Subfamily lentivirinae; System; Techniques; Testing; Tissues; Toxicology; Transfusion; Transgenes; Translational Research; Translations; Vascular blood supply; Viral; Virus; Work; abstracting; base; c-myc Genes; cell type; disease mechanisms study; embryonic stem cell; genetic manipulation; human subject; human tissue; induced pluripotent stem cell; infancy; isoimmunity; meetings; progenitor; protein aminoacid sequence; research study; retroviral transduction; stem; stem cell technology; tumor; vector

DESCRIPTION (provided by applicant): The limited supply of red blood cells (RBC) and platelets is a serious medical issue that can have life-threatening consequences for transfusion-dependent patients, particularly those who develop platelet allo-immunity. The low prevalence of O Rh-negative 'universal donor' blood type in the general population (<8% in Western countries and <0.3% in Asia) further intensifies the consequences of blood shortages for emergency situations where blood supply and/or typing is limited. Based on the pioneering work of Yamanaka and Thomson, human induced pluripotent stem cells (hiPSCs) offer the possibility to generate patient-specific stem cells without destruction of embryos. However, current iPSCs suffer from major drawbacks including multiple viral integrations and remaining transgenes at various chromosomal locations, any of which may cause unpredictable genetic dysfunction and/or tumor formation (Yamanaka, 2009). In this proposal, based on our recent "proof-of-concept" results, we hypothesize that hiPS cells suitable for clinical translation can be generated by direct delivery of reprogramming proteins attached to a cell penetrating peptide. In particular, we will generate fully reprogrammed hiPS cell lines from both healthy and O(-) subjects by direct protein delivery and test whether these cells can be propagated and expanded in vitro indefinitely, thus providing a potentially inexhaustible and donor-less source of blood lineage cells. This proposal will fully optimize the protein reprogramming methods to establish a highly efficient and safe way of reprogramming human tissues without genetic manipulation, and will address whether these hiPS cells can be used as a personalized platelet source and/or universal RBC source. PUBLIC HEALTH RELEVANCE: Although still in its infancy, "induced pluripotent stem cell (iPSC)" technology has the potential to revolutionize biomedical research, disease mechanism studies, and customized cell-based therapies. To explore the potential of iPSC's as a source of universal red blood cells and personalized platelets, we propose to establish and characterize clinically viable iPSC lines from healthy O Rh-negative [O(-)] subjects by direct delivery of reprogramming proteins without the use of viruses or foreign-DNA vectors. Using these protein-induced iPS cells, we will address whether they can be propagated and expanded in vitro indefinitely into blood cell progenitor's lineages that can be used as a donor-less source of universal red blood cells and/or personalized platelets.

描述（由申请人提供）：红细胞（RBC）和血小板的供应有限是一个严重的医学问题，可能对输血依赖患者造成危及生命的后果，特别是那些发生血小板同种免疫的患者。普通人群中O Rh阴性“万能供体”血型的低流行率（西方国家<8%，亚洲<0.3%）进一步加剧了血液供应和/或血型定型有限的紧急情况下血液短缺的后果。基于Yamanaka和Thomson的开创性工作，人类诱导多能干细胞（hiPSC）提供了在不破坏胚胎的情况下产生患者特异性干细胞的可能性。然而，目前的iPSC存在主要缺点，包括多个病毒整合和在不同染色体位置的剩余转基因，其中任何一个都可能导致不可预测的遗传功能障碍和/或肿瘤形成（Yamanaka，2009）。在这个提议中，基于我们最近的“概念验证”结果，我们假设可以通过直接递送附着于细胞穿透肽的重编程蛋白来产生适合于临床翻译的hiPS细胞。特别是，我们将通过直接蛋白质递送从健康和O（-）受试者中产生完全重编程的hiPS细胞系，并测试这些细胞是否可以在体外无限期地繁殖和扩增，从而提供潜在的取之不尽且无供体的血液谱系细胞来源。该提案将充分优化蛋白质重编程方法，以建立一种高效安全的方法来重编程人体组织，而无需遗传操作，并将解决这些hiPS细胞是否可以用作个性化血小板来源和/或通用RBC来源。 公共卫生相关性：虽然仍处于起步阶段，但“诱导多能干细胞（iPSC）”技术有可能彻底改变生物医学研究，疾病机制研究和定制的基于细胞的疗法。为了探索iPSC作为通用红细胞和个性化血小板来源的潜力，我们提出通过直接递送重编程蛋白而不使用病毒或外源DNA载体来建立和表征来自健康O Rh阴性[O（-）]受试者的临床上可行的iPSC系。使用这些蛋白诱导的iPS细胞，我们将解决它们是否可以在体外无限繁殖和扩增成血细胞祖细胞谱系，这些谱系可以用作通用红细胞和/或个性化血小板的无供体来源。

项目成果

Scalable generation of universal platelets from human induced pluripotent stem cells.

• DOI: 10.1016/j.stemcr.2014.09.010
• 发表时间: 2014-11-11
• 期刊: STEM CELL REPORTS
• 影响因子: 5.9
• 作者: Feng, Qiang;Shabrani, Namrata;Thon, Jonathan N.;Huo, Hongguang;Thiel, Austin;Machlus, Kellie R.;Kim, Kyungho;Brooks, Julie;Li, Feng;Luo, Chenmei;Kimbrel, Erin A.;Wang, Jiwu;Kim, Kwang-Soo;Italiano, Joseph;Cho, Jaehyung;Lu, Shi-Jiang;Lanza, Robert
• 通讯作者: Lanza, Robert

Transcription elongation factor Tcea3 regulates the pluripotent differentiation potential of mouse embryonic stem cells via the Lefty1-Nodal-Smad2 pathway.

• DOI: 10.1002/stem.1284
• 发表时间: 2013-02
• 期刊: STEM CELLS
• 影响因子: 5.2
• 作者: Park, Kyung-Soon;Cha, Young;Kim, Chun-Hyung;Ahn, Hee-Jin;Kim, Dohoon;Ko, Sanghyeok;Kim, Kyeoung-Hwa;Chang, Mi-Yoon;Ko, Jong-Hyun;Noh, Yoo-Sun;Han, Yong-Mahn;Kim, Jonghwan;Song, Jihwan;Kim, Jin Young;Tesar, Paul J.;Lanza, Robert;Lee, Kyung-Ah;Kim, Kwang-Soo
• 通讯作者: Kim, Kwang-Soo