Walter S. and Lucienne Driskill
Graduate Training Program in Life Sciences

DGP Northwestern

John Crispino

John Crispino, PhD Associate Professor Medicine Division of Hematology/Oncology Transcriptional regulation of normal and malignant blood cell development Research clusters: Cancer Biology Developmental Systems and Stem Cell Biology Genetics and Genomics E-mail:   j-crispino@northwestern.edu

Research in the Crispino lab is focused on investigating the regulatory mechanisms governing normal and malignant blood cell development, with an emphasis on understanding the growth of erythroid cells (red blood cells) and megakaryocytes (platelet-producing cells). In addition, we are greatly interested in learning how changes in normal essential regulatory molecules lead to human blood diseases, including leukemias, myelodysplastic syndromes (MDS) and myeloproliferative neoplasms (MPNs). Areas of major interest include:

Mechanisms of leukemogenesis in children with Down syndrome
Several years ago, we discovered that the zinc finger transcription factor GATA-1 is mutated in Acute Megakaryoblastic Leukemia (AMKL) in children with Down syndrome (DS). We further found that GATA1 mutations are also present in a pre-leukemia, named transient myeloproliferative disorder, which is relatively common in infants with Down syndrome. Current studies in the lab are focused on characterizing the role of GATA-1 in normal megakaryocyte development and on investigating how GATA1 mutations contribute to leukemia. In related experiments, we are also studying the mechanisms by which trisomy 21 promotes the development of leukemia. Our long-term goal is to unravel the mystery of why children with DS are predisposed to leukemia. In addition to studying mouse models of Down syndrome, we are now harnessing the power of induced pluripotent stem (iPS) cells to define the role of trisomy 21 in leukemia. 

Regulation of megakaryocyte proliferation, polyploidization and terminal maturation:
Megakaryocytes, the precursors to platelets, are one of the few cells types to undergo polyploidization in the normal course of maturation. We are using high throughput methods to identify genes that control the switch between the proliferative and the endomitotic cell cycle. In addition, we are examining the contribution of key transcription factors, such as GATA-1, GATA-2 and Ets proteins in the regulation of megakaryocyte gene expression, terminal differentiation and lineage commitment.

Development of novel therapeutics for human megakaryocytic neoplasms:
In an ongoing collaboration with the Broad Institute, we have identified a set of small molecules that rapidly and robustly induce proliferation arrest and terminal maturation of malignant megakaryocytes, including those that harbor mutations in GATA1, JAK3, or c-MPL, as well as those that express the t(1;22) fusion protein associated with pediatric non-DS AMKL. These small molecules induce features of megakaryocyte differentiation, including extensive polyploidization and expression of markers of terminal differentiation, but they do not promote platelet release. We are currently developing these compounds as potential new therapies for blood disorders, including acute megakaryocytic leukemia and primary myelofibrosis. 

Control of red blood cells development:
Survivin, a member of the inhibitor of apoptosis (IAP) family that also plays an essential role in cytokinesis, is differentially expressed during erythroid versus megakaryocyte development. We recently reported that survivin is required for the maintenance of hematopoietic stem and progenitor cells as well as for red blood cell development. More recently, we have found that survivin is highly expressed in post-mitotic erythroblasts. We are currently studying the contribution of survivin to terminal maturation of erythroid cells, including enucleation.

Selected Publications:
Muntean, AG, Pang, L, Poncz, M, Dowdy, S, Blobel, G, and Crispino, JD. (2007) Cyclin D-Cdk4 is regulated by GATA-1 and required for megakaryocyte growth and polyploidization. Blood 109:5199-5207. 

Leung, CG, Xu, Y, Mularski, B, Liu, H, Gurbuxani, S, and Crispino, JD. (2007) Requirements for Survivin in terminal differentiation of erythroid cells and maintenance of hematopoietic stem and progenitor cells. Journal of Experimental Medicine 204:1603-1611. 

Huang, Z, Richmond, TD, Muntean, A, Barber, DL, Weiss, MJ, and Crispino, JD. (2007) STAT1 promotes megakaryopoiesis downstream of GATA-1. Journal of Clinical Investigation 117:3890-3899. 

Kirsammer, G, Jilani, S, Liu, H, Davis, E, Gurbuxani, S, Le Beau, MM, and Crispino, JD. (2008) Highly penetrant myeloproliferative disease in the Ts65Dn mouse model of Down syndrome. Blood, 111:767-775. 

McCrann, DJ, Yezefski T, Nguyen HG, Liu H, Wen Q, Crispino JD, and Ravid K. (2008) Survivin overexpression in transgenic mice alone does not alter megakaryocyte ploidy nor interfere with erythroid/megakaryocytic lineage development.  Blood 101:4092-4095. 

Malinge, S, Izraeli, S, and Crispino JD. (2009) Insights into the manifestations, outcomes and mechanisms of leukemogenesis in Down syndrome. Blood 113:2619-2628.

Stankiewicz, MJ, and Crispino JD. (2009) ETS2 and ERG promote megakaryopoiesis and synergize with loss of GATA-1 to immortalize hematopoietic progenitor cells. Blood 113:3337-3347.  

Wen, Q, Leung, C, Huang, Z, Small, S, Reddi, AL, Licht, JD and Crispino JD. (2009) Survivin is not required for the endomitotic cell cycle of megakaryocytes. Blood 114:153-156.

Abdel-Wahab, O., Mullally, A, Hedvat, C, Garcia-Maero, G, Patel, J, Malinge S, Yao, J, Kilipavaara, O, Wadleigh, M, Bhat, R, Huberman, K, Thomas, S, Dolgalev, I, Heguy, A, Paietta, E, Le Beau, M, Beran, M, Tallman, M, Kantarjian, H, Ebert, B, Gilliland, DG, Crispino, JD, and Levine, R. (2009) Genetic characteristics of TET1, TET2, and TET3 alterations in myeloid malignancies. Blood 114:144-147.

Bhat, R, Malinge, S, Gamis, AS, Sorrell, AD, Hilden, JM, Paietta, E, Tallman, MS, Ketterling, RP, and Crispino, JD. Mutational analysis of candidate tumor associated genes in acute megakaryoblastic leukemia. Leukemia, in press.

Huang, Z, Dore, LC, Li, Z, Orkin, SH, Feng, G, Lin, S, and Crispino JD. GATA2 reinforces megakaryocyte development in the absence of GATA-1. MCB, in press.