Asmin Tulpule

Dana-Farber Cancer Institute, Boston, Massachusetts, United States

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Publications (5)51.79 Total impact

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    James M Kelley · Asmin Tulpule · George Q Daley
    Cell cycle (Georgetown, Tex.) 10/2013; 12(22). DOI:10.4161/cc.26698 · 4.57 Impact Factor
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    ABSTRACT: Shwachman-Diamond syndrome (SDS), a rare autosomal-recessive disorder characterized by exocrine pancreatic insufficiency and hematopoietic dysfunction, is caused by mutations in the Shwachman-Bodian-Diamond syndrome (SBDS) gene. We created human pluripotent stem cell models of SDS through knockdown of SBDS in human embryonic stem cells (hESCs) and generation of induced pluripotent stem cell (iPSC) lines from two patients with SDS. SBDS-deficient hESCs and iPSCs manifest deficits in exocrine pancreatic and hematopoietic differentiation in vitro, enhanced apoptosis, and elevated protease levels in culture supernatants, which could be reversed by restoring SBDS protein expression through transgene rescue or by supplementing culture media with protease inhibitors. Protease-mediated autodigestion provides a mechanistic link between the pancreatic and hematopoietic phenotypes in SDS, highlighting the utility of hESCs and iPSCs in obtaining novel insights into human disease.
    Cell stem cell 04/2013; 12(6). DOI:10.1016/j.stem.2013.04.002 · 22.27 Impact Factor
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    ABSTRACT: Fanconi anemia (FA) is a genetically heterogeneous, autosomal recessive disorder characterized by pediatric bone marrow failure and congenital anomalies. The effect of FA gene deficiency on hematopoietic development in utero remains poorly described as mouse models of FA do not develop hematopoietic failure and such studies cannot be performed on patients. We have created a human-specific in vitro system to study early hematopoietic development in FA using a lentiviral RNA interference (RNAi) strategy in human embryonic stem cells (hESCs). We show that knockdown of FANCA and FANCD2 in hESCs leads to a reduction in hematopoietic fates and progenitor numbers that can be rescued by FA gene complementation. Our data indicate that hematopoiesis is impaired in FA from the earliest stages of development, suggesting that deficiencies in embryonic hematopoiesis may underlie the progression to bone marrow failure in FA. This work illustrates how hESCs can provide unique insights into human development and further our understanding of genetic disease.
    Blood 04/2010; 115(17):3453-62. DOI:10.1182/blood-2009-10-246694 · 10.45 Impact Factor
  • Asmin Tulpule · George Q Daley
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    ABSTRACT: Human embryonic stem cells (hESCs) represent a powerful platform to study human development and its dysfunction in human disease. However, certain biological properties have hampered the application of standard gain of function and loss of function tools to these cells. For example, while traditional gene knockouts by homologous recombination (HR) have been reported, the low cloning efficiency of hESCs has made HR a lengthy and laborious undertaking. An alternative method of achieving loss of function is the use of small interfering RNAs (siRNAs) that can be introduced either as pre-synthesized duplexed oligonucleotides or via lentiviral vector. The use of a lentiviral vector to deliver siRNAs has proven to be a rapid and specific way to achieve highly efficient and persistent gene knockdowns in hESCs. In this chapter, we will summarize the key requirements for the successful application of lentiviral RNAi in hESCs.
    Methods in Molecular Biology 02/2009; 482:35-42. DOI:10.1007/978-1-59745-060-7_3 · 1.29 Impact Factor
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    ABSTRACT: Deficiencies in the SBDS gene result in Shwachman-Diamond syndrome (SDS), an inherited bone marrow failure syndrome associated with leukemia predisposition. SBDS encodes a highly conserved protein previously implicated in ribosome biogenesis. Using human primary bone marrow stromal cells (BMSCs), lymphoblasts, and skin fibroblasts, we show that SBDS stabilized the mitotic spindle to prevent genomic instability. SBDS colocalized with the mitotic spindle in control primary BMSCs, lymphoblasts, and skin fibroblasts and bound to purified microtubules. Recombinant SBDS protein stabilized microtubules in vitro. We observed that primary BMSCs and lymphoblasts from SDS patients exhibited an increased incidence of abnormal mitoses. Similarly, depletion of SBDS by siRNA in human skin fibroblasts resulted in increased mitotic abnormalities and aneuploidy that accumulated over time. Treatment of primary BMSCs and lymphoblasts from SDS patients with nocodazole, a microtubule destabilizing agent, led to increased mitotic arrest and apoptosis, consistent with spindle destabilization. Conversely, SDS patient cells were resistant to taxol, a microtubule stabilizing agent. These findings suggest that spindle instability in SDS contributes to bone marrow failure and leukemogenesis.
    Journal of Clinical Investigation 05/2008; 118(4):1511-8. DOI:10.1172/JCI33764 · 13.22 Impact Factor