Human embryonic stem (ES) and embryonic germ (EG) cell lines provide unprecedented opportunities to study self-renewal and differentiation of embryonic and adult stem cells, and to develop novel cell-based clinical therapies. Emerging data has revealed that human ES/EG cells are pluripotent like their mouse counterparts, but also distinct in many critical properties of cell proliferation and differentiation. Therefore, it is necessary to study directly human ES/EG cells and their differentiation. We have begun to use human ES/EG cells and their immediate progeny, embryoid body-derived (EBD) cells, for hematopoietic differentiation, building upon our previous studies with mouse ES and EG cells. With the H1 (WA01) human ES (hES) cell line, we have made significant progress in 3 areas: 1) developed a method to generate both lymphoid (B and NK) and myeloid cells from differentiated hES cells; 2) developed a method to efficiently and stably transduce hES cells by lentiviral vectors; 3) developed a culture system to expand hES cells on human marrow stromal cells (hMSCs) in replacing previously required mouse feeder cells. Therefore, we planned a 2nd stage project to improve and elucidate mechanisms of lympho-hematopoietic differentiation from hES cells and non-tumorgeneic EBD cell lines. The overall goal is to produce transplantable lympho-hematopoietic progenitors capable of engrafting in NOD/SCID mice. In addition to using hMSCs as stroma, we will also use lentiviral vectors to express key regulatory genes such as VEGF, Notch as well as HoxB4 in hES cells to further facilitate self-renewal and engraftment of the generated lympho-hematopoietic progenitors. These approaches will allow us to define external/internal signals required for the genesis and self-renewal of human hematopoietic stem cells. Later on, we will directly examine the generation and functionality of antigen-presenting cells (APCs) which express high levels of MHC class II complex and regulate T cells. Using APCs derived from hES and EBD cells (+/- gene modification), we will attempt to inactivate alloreactive T cells and explore strategies ultimately leading to immune tolerance induction. This research project will allow us to better understand early events of human lympho-hematopoiesis and human stem cells. It will also provide a foundation for developing novel ES cell-based therapies that require reconstituting or re-programming patient's blood/immune systems.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL073781-02
Application #
6798184
Study Section
Special Emphasis Panel (ZHL1-CSR-J (M2))
Program Officer
Thomas, John
Project Start
2003-09-01
Project End
2009-08-31
Budget Start
2004-09-01
Budget End
2005-08-31
Support Year
2
Fiscal Year
2004
Total Cost
$408,750
Indirect Cost
Name
Johns Hopkins University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Liu, Yanfeng; Wang, Ying; Gao, Yongxing et al. (2015) Efficient generation of megakaryocytes from human induced pluripotent stem cells using food and drug administration-approved pharmacological reagents. Stem Cells Transl Med 4:309-19
Smith, Cory; Abalde-Atristain, Leire; He, Chaoxia et al. (2015) Efficient and allele-specific genome editing of disease loci in human iPSCs. Mol Ther 23:570-7
Chou, Bin-Kuan; Gu, Haihui; Gao, Yongxing et al. (2015) A facile method to establish human induced pluripotent stem cells from adult blood cells under feeder-free and xeno-free culture conditions: a clinically compliant approach. Stem Cells Transl Med 4:320-32
Huang, Xiaosong; Wang, Ying; Yan, Wei et al. (2015) Production of Gene-Corrected Adult Beta Globin Protein in Human Erythrocytes Differentiated from Patient iPSCs After Genome Editing of the Sickle Point Mutation. Stem Cells 33:1470-9
Huang, Xiaosong; Shah, Siddharth; Wang, Jing et al. (2014) Extensive ex vivo expansion of functional human erythroid precursors established from umbilical cord blood cells by defined factors. Mol Ther 22:451-463
Huang, Kevin; Shen, Yin; Xue, Zhigang et al. (2014) A panel of CpG methylation sites distinguishes human embryonic stem cells and induced pluripotent stem cells. Stem Cell Reports 2:36-43
Cai, Jun; Miao, Xuexia; Li, Yueying et al. (2014) Whole-genome sequencing identifies genetic variances in culture-expanded human mesenchymal stem cells. Stem Cell Reports 3:227-33
Wang, Ying; Cheng, Linzhao; Gerecht, Sharon (2014) Efficient and scalable expansion of human pluripotent stem cells under clinically compliant settings: a view in 2013. Ann Biomed Eng 42:1357-72
Shah, Siddharth; Huang, Xiaosong; Cheng, Linzhao (2014) Concise review: stem cell-based approaches to red blood cell production for transfusion. Stem Cells Transl Med 3:346-55
Smith, Cory; Gore, Athurva; Yan, Wei et al. (2014) Whole-genome sequencing analysis reveals high specificity of CRISPR/Cas9 and TALEN-based genome editing in human iPSCs. Cell Stem Cell 15:12-3

Showing the most recent 10 out of 41 publications