Abstract

Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired blood disease characterized by chronic hemolysis causing hemoglobinuria, bone marrow (BMF) failure, and a tendency to thrombosis. PNH is due to the expansion of an abnormal done which arises through a somatic mutation in the X- linked PIG-A gene on the active X-chromosome in an early hematopoietic stem cell. The PIG-A mutation abolishes or impairs the biosynthesis of glycosyl phosphatidylinositol (GPI) anchors. Cells belonging to the FNH done are therefore deficient in all proteins that are anchored to the membrane through GPI anchors. PNH is closely related to aplastic anemia (AA), in that the incidence of PNH is increased in patients with AA, and AA is often a dreaded end-stage of PNH. This proposal aims to test the working hypothesis that: (a) PNH is always associated with some degree of BMF; (b) hematopoietic clones with the PNH phenotype (GPI-negative) have a survival and/or growth advantage under conditions that cause or maintain BMF. To test this hypothesis the plan is to create an animal model, in which some or all of hematopoiesis lacks all GPI-linked proteins. For this purpose inactivation of the PIG-A gene in mouse embryonic stem (ES) cells has been already achieved. These 'PIG-A-null' ES cells will now be used in order to study their differentiation in-vitro, and their ability to contribute to mouse development after transfer into blastocysts. Since the phenotype of a fully PIG-A null animal might be severe or not compatible with life, several different approaches will be explored to achieve viable PNH hematopoiesis. (1) Rescue of mutants with impaired hematopoiesis (e.g. W/W) by transfer of PIG-A null ES cells into mutant blastocysts. (2) Transplantation of hematopoietic cells derived from PIG-A-null ES-cell into irradiated recipients. Finally, PIG-A positive and PIG-A null hematopoiesis will be compared within the same animal under normal conditions, as well as under conditions that cause or maintain BMF.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL056778-04
Application #
6043923
Study Section
Hematology Subcommittee 2 (HEM)
Project Start
1996-08-01
Project End
2000-07-31
Budget Start
1999-08-01
Budget End
2000-07-31
Support Year
4
Fiscal Year
1999
Total Cost
Indirect Cost

  Institution

Name
Sloan-Kettering Institute for Cancer Research
Department
Type
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10065

  Publications

Araten, David J; Luzzatto, Lucio (2006) The mutation rate in PIG-A is normal in patients with paroxysmal nocturnal hemoglobinuria (PNH). Blood 108:734-6
Ellis, Nathan A; Kirchhoff, Tomas; Mitra, Nandita et al. (2006) Localization of breast cancer susceptibility loci by genome-wide SNP linkage disequilibrium mapping. Genet Epidemiol 30:48-61
Araten, David J; Thaler, Howard T; Luzzatto, Lucio (2005) High incidence of thrombosis in African-American and Latin-American patients with Paroxysmal Nocturnal Haemoglobinuria. Thromb Haemost 93:88-91
Araten, David J; Golde, David W; Zhang, Rong H et al. (2005) A quantitative measurement of the human somatic mutation rate. Cancer Res 65:8111-7
Mitra, Nandita; Ye, Tian-Zhang; Smith, Alex et al. (2004) Localization of cancer susceptibility genes by genome-wide single-nucleotide polymorphism linkage-disequilibrium mapping. Cancer Res 64:8116-25
Karadimitris, Anastasios; Araten, David J; Luzzatto, Lucio et al. (2003) Severe telomere shortening in patients with paroxysmal nocturnal hemoglobinuria affects both GPI- and GPI+ hematopoiesis. Blood 102:514-6
Bessler, Monica; Rosti, Vittorio; Peng, Yufeng et al. (2002) Glycosylphosphatidylinositol-linked proteins are required for maintenance of a normal peripheral lymphoid compartment but not for lymphocyte development. Eur J Immunol 32:2607-16
Araten, D J; Bessler, M; McKenzie, S et al. (2002) Dynamics of hematopoiesis in paroxysmal nocturnal hemoglobinuria (PNH): no evidence for intrinsic growth advantage of PNH clones. Leukemia 16:2243-8
Karadimitris, A; Li, K; Notaro, R et al. (2001) Association of clonal T-cell large granular lymphocyte disease and paroxysmal nocturnal haemoglobinuria (PNH): further evidence for a pathogenetic link between T cells, aplastic anaemia and PNH. Br J Haematol 115:1010-4
Araten, D J; Swirsky, D; Karadimitris, A et al. (2001) Cytogenetic and morphological abnormalities in paroxysmal nocturnal haemoglobinuria. Br J Haematol 115:360-8

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