John H. Bushweller

Professor, Molecular Physiology and Biological Physics

Primary Location

PO Box 800736

Primary Phone Number

434.243.6409

[email protected]

Website

http://www.bushwellerlab.org/

Bio & Overview

Drug Development Targeting Transcription Drivers in Cancer Dysregulation of gene expression is a hallmark of all cancers. It is critical for self-renewal and chemo-resistance of cancer cells which contributes to the inability to completely eradicate cancer cells, thereby leading to relapse. The specific gene expression program that confers these properties derives from the aberrant activity of specific transcription factors which are drivers of disease. Clearly, the most direct and effective approach to alter this gene expression program is to directly target the activity of the transcription factors driving the disease. There are numerous examples of such transcription factor drivers in cancer such as fusion proteins of RUNX1 and CBFβ in leukemia, fusion proteins of ERG in prostate cancer and Ewing’s sarcoma, ETV-1 in melanoma, other members of the Ets family of transcription factors in a variety of different cancers, etc. Transcription factors have traditionally been viewed as “undruggable” due to the need to target more challenging protein-protein or protein nucleic acid interactions through which these proteins act. There are still relatively few examples of such agents for cancer treatment, with the MDM2-p53 inhibitors being one example of such an agent that has progressed to the clinic. We are exploring several unique approaches to target this important class of proteins for drug development. Illendula A, Pulikkan JA, Zong H, Grembecka J, Xue L, Sen S, Zhou Y, Boulton A, Kuntimaddi A, Gao Y, Rajewski RA, Guzman ML, Castilla LH, Bushweller JH. Chemical biology. A small-molecule inhibitor of the aberrant transcription factor CBFβ-SMMHC delays leukemia in mice. Science. 2015 Feb 13;347(6223):779-84. PubMed PMID: 25678665; PubMed Central PMCID: PMC4423805. Illendula A, Gilmour J, Grembecka J, Tirumala VSS, Boulton A, Kuntimaddi A, Schmidt C, , Wang L, John A. Pulikkan, Hongliang Zong, Mahmut Parlak, Cem Kuscu, Anna Pickin, Yunpeng Zhou, Yan Gao Y, Mishra L, Adli M, Castilla LH, Rajewski RA, Janes KA, Guzman ML, Bonifer C, and Bushweller JH. Small Molecule Inhibitor of CBFB-RUNX Binding for RUNX Transcription Factor Driven Cancers. EBioMedicine. 2016 8: 117-131. Pubmed PMID: 27428424; PubMed Central PMCID: PMC4919611. Pulikkan JA, Hegde M, Belaghzal H, Illendula A, Yu J, Ahmed H, O’Hagan K, Ou J, Muller-Tidow C, Wolfe SA, Zhu LJ, Dekker J, Bushweller JH, Castilla LH. CBFβ-SMMHC inhibition leads to alteration of chromatin dynamics at MYC distal enhancers and abrogation of inv(16) leukemia. Cell. 2018 174(1):172-186.e21. Pubmed PMID: 29958106. Structure/Function Studies of Transcription Factor Drivers in Cancer Our lab is fundamentally interested in understanding, from a structural and biophysical perspective, the functioning of proteins involved in regulating transcription, particularly those involved in the dysregulation associated with the development of cancer. Structural and functional characterization of the native forms of these proteins and their relevant complexes via NMR spectroscopy, X-ray crystallography, and a variety of other techniques provides a baseline of understanding. Subsequent characterization of the oncoprotein forms then provides a detailed understanding of the molecular mechanism of oncogenesis associated with altered forms of these proteins. Such knowledge leads to novel avenues for the design of therapeutic agents to treat the cancers associated with these particular oncoproteins. A long-term focus has been structural studies of a novel transcription factor referred to as the core-binding factor (CBF). This heterodimeric protein is essential for hematopoietic development. Gene translocations associated with the genes coding for the two subunits of CBF produce novel fusion proteins which have been implicated as playing a role in more than 30% of acute leukemias. We have carried out extensive structural and functional studies of the oncoprotein forms of the two subunits of CBF that are associated with leukemia. We have extended these studies to the MLL protein, a key epigenetic regulator that is the target of chromosomal translocations in leukemia which are particularly poor prognosis. Cierpicki T, Risner LE, Grembecka J, Lukasik SM, Popovic R, Omonkowska M, Shultis DD, Zeleznik-Le NJ, Bushweller JH. Structure of the MLL CXXC domain-DNA complex and its functional role in MLL-AF9 leukemia. Nat Struct Mol Biol. 2010 Jan;17(1):62-8. PubMed PMID: 20010842; PubMed Central PMCID: PMC2908503. Kuntimaddi A, Achille NJ, Thorpe J, Lokken AA, Singh R, Hemenway CS, Adli M, Zeleznik-Le NJ, Bushweller JH. Degree of recruitment of DOT1L to MLL-AF9 defines level of H3K79 Di- and tri-methylation on target genes and transformation potential. Cell Rep. 2015 May 5;11(5):808-20. PubMed PMID: 25921540; PubMed Central PMCID: PMC4426023. List of Publications in Pubmed: "https://www.ncbi.nlm.nih.gov/pubmed/?term=Bushweller+JH"

Academic Information

Department: Molecular Physiology and Biological Physics

Academic Title: Professor

Primary Education

BA, Chemistry, Dartmouth College, 1984
PhD, Chemistry, University of California, Berkeley, CA, 1990
Postdoc, undefinedEidgenössische Technische Hochschule, Zürich, Switzerland, 1993

Other Appointments

Professor, Chemistry
Pharmacology
Professor, BMBG
Professor, SCBB
Professor, MOMD

Other Affiliations

Program Leader, Cancer Center
Professor, Arts & Sciences
Program Co-Leader, Chemical and Structural Biology Program
MSTP
Professor, BIMS

Research Interests

Drug Development Targeting Transcription Drivers in Cancer; Structure/Function Studies of Transcription Factor Drivers in Cancer

Research Disciplines

Biochemistry
Biophysics
Biophysics & Structural Biology
Cancer Biology
Chemistry
Epigenetics
Molecular Pharmacology
Physiology
Translational Science

Research Opportunities

Biophysics & Structural Biology
Cancer Biology
Chemistry and Physics of Living Systems
Epigenetics
Molecular Pharmacology

PhD Programs

Biophysics
Physiology

Selected Publications

2024

Peramangalam, P. S., Surapally, S., Veltri, A. J., Zheng, S., Burns, R., Zhu, N., . . . Pulikkan, J. A. (2024). N-MYC regulates cell survival via eIF4G1 in inv(16) acute myeloid leukemia. SCIENCE ADVANCES, 10(9). doi:10.1126/sciadv.adh8493

2022

Wray, J. P., Deltcheva, E. M., Boiers, C., Chhetri, J. B., Brown, J., Gagrica, S., . . . Richardson, S. E. (2022). Regulome analysis in B-acute lymphoblastic leukemia exposes Core Binding Factor addiction as a therapeutic vulnerability. NATURE COMMUNICATIONS, 13(1). doi:10.1038/s41467-022-34653-3

Kalmode, H. P., Podsiadly, I., Kabra, A., Boulton, A., Reddy, P., Gao, Y., . . . Bushweller, J. H. (2022). Small-Molecule Inhibitors of the MLL1 CXXC Domain, an Epigenetic Reader of DNA Methylation. ACS MEDICINAL CHEMISTRY LETTERS, 13(8), 1363-1369. doi:10.1021/acsmedchemlett.2c00198

Kim, K. -B., Kabra, A., Kim, D. -W., Xue, Y., Huang, Y., Hou, P. -C., . . . Park, K. -S. (2022). KIX domain determines a selective tumor-promoting role for EP300 and its vulnerability in small cell lung cancer. SCIENCE ADVANCES, 8(7). doi:10.1126/sciadv.abl4618

2021

Yu, L., Myers, G., Ku, C. -J., Schneider, E., Wang, Y., Singh, S. A., . . . Engel, J. D. (2021). An erythroid-to-myeloid cell fate conversion is elicited by LSD1 inactivation. BLOOD, 138(18), 1691-1704. doi:10.1182/blood.2021011682

Kabra, A., & Bushweller, J. (2022). The Intrinsically Disordered Proteins MLLT3 (AF9) and MLLT1 (ENL)-Multimodal Transcriptional Switches With Roles in Normal Hematopoiesis, MLL Fusion Leukemia, and Kidney Cancer. JOURNAL OF MOLECULAR BIOLOGY, 434(1). doi:10.1016/j.jmb.2021.167117

O'Hare, M., Amarnani, D., Whitmore, H. A. B., An, M., Marino, C., Ramos, L., . . . Kim, L. A. (2021). Targeting Runt-Related Transcription Factor 1 Prevents Pulmonary Fibrosis and Reduces Expression of Severe Acute Respiratory Syndrome Coronavirus 2 Host Mediators. AMERICAN JOURNAL OF PATHOLOGY, 191(7), 1193-1208. doi:10.1016/j.ajpath.2021.04.006

2020

Whitmore, H. A. B., Amarnani, D., O'Hare, M., Delgado-Tirado, S., Gonzalez-Buendia, L., An, M., . . . Kim, L. A. (2021). TNF-α signaling regulates RUNX1 function in endothelial cells. FASEB JOURNAL, 35(2). doi:10.1096/fj.202001668R

Czub, M. P., Boulton, A. M., Rastelli, E. J., Tasker, N. R., Maskrey, T. S., Blanco, I. K., . . . Lazo, J. S. (2020). Structure of the Complex of an Iminopyridinedione Protein Tyrosine Phosphatase 4A3 Phosphatase Inhibitor with Human Serum Albumin. MOLECULAR PHARMACOLOGY, 98(6), 648-657. doi:10.1124/molpharm.120.000131

Schmidt, C. R., Achille, N. J., Kuntimaddi, A., Boulton, A. M., Leach, B. I., Zhang, S., . . . Bushweller, J. H. (2020). BCOR Binding to MLL-AF9 Is Essential for Leukemia via Altered EYA1, SIX, and MYC Activity. BLOOD CANCER DISCOVERY, 1(2), 162-177. doi:10.1158/2643-3230.BCD-20-0036

Rose, J. T., Moskovitz, E., Boyd, J. R., Gordon, J. A., Bouffard, N. A., Fritz, A. J., . . . Stein, G. S. (2020). Inhibition of the RUNX1-CBFβ transcription factor complex compromises mammary epithelial cell identity: a phenotype potentially stabilized by mitotic gene bookmarking.. Oncotarget, 11(26), 2512-2530. doi:10.18632/oncotarget.27637

Bushweller, J. H. (2020). Protein Disulfide Exchange by the Intramembrane Enzymes DsbB, DsbD, and CcdA. JOURNAL OF MOLECULAR BIOLOGY, 432(18), 5091-5103. doi:10.1016/j.jmb.2020.04.008

Fritz, A. J., Hong, D., Boyd, J., Kost, J., Finstaad, K. H., Fitzgerald, M. P., . . . Stein, G. S. (2020). RUNX1 and RUNX2 transcription factors function in opposing roles to regulate breast cancer stem cells. JOURNAL OF CELLULAR PHYSIOLOGY, 235(10), 7261-7272. doi:10.1002/jcp.29625

Czub, M. P., Boulton, A. M., Rastelli, E. J., Tasker, N. R., Maskrey, T. S., Blanco, I. K., . . . Lazo, J. S. (2020). Structure of the Complex of an Iminopyridinedione Protein Tyrosine Phosphatase 4A3 Phosphatase Inhibitor with Human Serum Albumin (vol 98, pg 648, 2020). MOLECULAR PHARMACOLOGY, 98(6), 781. Retrieved from https://www.webofscience.com/

2019

Bushweller, J. H. (2019). Targeting transcription factors in cancer - from undruggable to reality. NATURE REVIEWS CANCER, 19(11), 611-624. doi:10.1038/s41568-019-0196-7

Alegre, F., Ormonde, A. R., Godinez, D. R., Illendula, A., Bushweller, J. H., & Wittenburg, L. A. (2020). The interaction between RUNX2 and core binding factor beta as a potential therapeutic target in canine osteosarcoma. VETERINARY AND COMPARATIVE ONCOLOGY, 18(1), 52-63. doi:10.1111/vco.12526

Mill, C. P., Fiskus, W., DiNardo, C. D., Qian, Y., Raina, K., Rajapakshe, K., . . . Bhalla, K. N. (2019). RUNX1-targeted therapy for AML expressing somatic or germline mutation in RUNX1. BLOOD, 134(1), 59-73. doi:10.1182/blood.2018893982

Zhou, N., Gutierrez-Uzquiza, A., Zheng, X. Y., Chang, R., Vogl, D. T., Garfall, A. L., . . . Busino, L. (2019). RUNX proteins desensitize multiple myeloma to lenalidomide via protecting IKZFs from degradation. LEUKEMIA, 33(8), 2006-2021. doi:10.1038/s41375-019-0403-2

Rose, J., Boyd, J., Gordon, J., Kang, M., Moskovitz, E., Bouffard, N., . . . Zaidi, S. (2019). Mitotic gene bookmarking by RUNX1 contributes to stabilization of the normal mammary epithelial phenotype. doi:10.1101/511410

2018

Pulikkan, J. A., Hegde, M., Ahmad, H. M., Belaghzal, H., Illendula, A., Yu, J., . . . Castilla, L. H. (2018). CBFβ-SMMHC Inhibition Triggers Apoptosis by Disrupting MYC Chromatin Dynamics in Acute Myeloid Leukemia (vol 174, pg 172, 2018). CELL, 174(5), 1325. doi:10.1016/j.cell.2018.08.014

Pulikkan, J. A., Hegde, M., Ahmad, H. M., Belaghzal, H., Illendula, A., Yu, J., . . . Castilla, L. H. (2018). CBFβ-SMMHC Inhibition Triggers Apoptosis by Disrupting MYC Chromatin Dynamics in Acute Myeloid Leukemia. CELL, 174(1), 172-+. doi:10.1016/j.cell.2018.05.048

Carlton, A. L., Illendula, A., Gao, Y., Llaneza, D. C., Boulton, A., Shah, A., . . . Bushweller, J. H. (2018). Small molecule inhibition of the CBFβ/RUNX interaction decreases ovarian cancer growth and migration through alterations in genes related to epithelial-to-mesenchymal transition. GYNECOLOGIC ONCOLOGY, 149(2), 350-360. doi:10.1016/j.ygyno.2018.03.005

Zhou, Y., & Bushweller, J. H. (2018). Solution structure and elevator mechanism of the membrane electron transporter CcdA. NATURE STRUCTURAL & MOLECULAR BIOLOGY, 25(2), 163-+. doi:10.1038/s41594-018-0022-z

2017

Oo, Z. M., Illendula, A., Grembecka, J., Schmidt, C., Zhou, Y., Esain, V., . . . Bushweller, J. H. (2018). A tool compound targeting the core binding factor Runt domain to disrupt binding to CBF in leukemic cells. LEUKEMIA & LYMPHOMA, 59(9), 2188-2200. doi:10.1080/10428194.2017.1410882

Illendula, A., Gilmour, J., Grembecka, J., Tirumala, V. S. S., Boulton, A., Kuntimaddi, A., . . . Bushweller, J. H. (2017). Small Molecule Inhibitor of CBFβ-RUNX Binding for RUNX Transcription Factor Driven Cancers (vol 8, pg 117, 2016). EBIOMEDICINE, 25, 188. doi:10.1016/j.ebiom.2017.10.029

Choi, A., Illendula, A., Pulikkan, J. A., Roderick, J. E., Tesell, J., Yu, J., . . . Kelliher, M. A. (2017). RUNX1 is required for oncogenic Myb and Myc enhancer activity in T-cell acute lymphoblastic leukemia. BLOOD, 130(15), 1722-1733. doi:10.1182/blood-2017-03-775536

Margulieux, K. R., Liebov, B. K., Tirumala, V. S. K. K. S., Singh, A., Bushweller, J. H., Nakamoto, R. K., & Hughes, M. A. (2017). Bacillus anthracis Peptidoglycan Integrity Is Disrupted by the Chemokine CXCL10 through the FtsE/X Complex. FRONTIERS IN MICROBIOLOGY, 8. doi:10.3389/fmicb.2017.00740

Tahirov, T. H., & Bushweller, J. (2017). Structure and Biophysics of CBFβ/RUNX and Its Translocation Products. RUNX PROTEINS IN DEVELOPMENT AND CANCER, 962, 21-31. doi:10.1007/978-981-10-3233-2_2

Castilla, L. H., & Bushweller, J. H. (2017). Molecular Basis and Targeted Inhibition of CBFβ-SMMHC Acute Myeloid Leukemia. RUNX PROTEINS IN DEVELOPMENT AND CANCER, 962, 229-244. doi:10.1007/978-981-10-3233-2_15

2016

Illendula, A., Gilmour, J., Grembecka, J., Tirumala, V. S. S., Boulton, A., Kuntimaddi, A., . . . Bushweller, J. H. (2016). Small Molecule Inhibitor of CBFβ-RUNX Binding for RUNX Transcription Factor Driven Cancers. EBIOMEDICINE, 8, 117-131. doi:10.1016/j.ebiom.2016.04.032

Huang, Y., Thoms, J. A. I., Tursky, M. L., Knezevic, K., Beck, D., Chandrakanthan, V., . . . Pimanda, J. E. (2016). MAPK/ERK2 phosphorylates ERG at serine 283 in leukemic cells and promotes stem cell signatures and cell proliferation. LEUKEMIA, 30(7), 1552-1561. doi:10.1038/leu.2016.55

2015

Xiao, S., Brannon, M. K., Zhao, X., Fread, K. I., Ellena, J. F., Bushweller, J. H., . . . Capelluto, D. G. S. (2015). Tom1 Modulates Binding of Tollip to Phosphatidylinositol 3-Phosphate via a Coupled Folding and Binding Mechanism. STRUCTURE, 23(10), 1910-1920. doi:10.1016/j.str.2015.07.017

Kuntimaddi, A., Achille, N. J., Thorpe, J., Lokken, A. A., Singh, R., Hemenway, C. S., . . . Bushweller, J. H. (2015). Degree of Recruitment of DOT1L to MLL-AF9 Defines Level of H3K79 Di- and Tri-methylation on Target Genes and Transformation Potential. CELL REPORTS, 11(5), 808-820. doi:10.1016/j.celrep.2015.04.004

Illendula, A., Pulikkan, J. A., Zong, H., Grembecka, J., Xue, L., Sen, S., . . . Bushweller, J. H. (2015). A small-molecule inhibitor of the aberrant transcription factor CBFβ-SMMHC delays leukemia in mice. SCIENCE, 347(6223), 779-784. doi:10.1126/science.aaa0314

2014

Lokken, A. A., Achille, N. J., Chang, M. -J., Lin, J. J., Kuntimaddi, A., Leach, B. I., . . . Hemenway, C. S. (2014). Importance of a specific amino acid pairing for murine MLL leukemias driven by MLLT1/3 or AFF1/4. LEUKEMIA RESEARCH, 38(11), 1309-1315. doi:10.1016/j.leukres.2014.08.010

2013

Risner, L. E., Kuntimaddi, A., Lokken, A. A., Achille, N. J., Birch, N. W., Schoenfelt, K., . . . Zeleznik-Le, N. J. (2013). Functional Specificity of CpG DNA-binding CXXC Domains in Mixed Lineage Leukemia. JOURNAL OF BIOLOGICAL CHEMISTRY, 288(41), 29901-29910. doi:10.1074/jbc.M113.474858

Regan, M. C., Horanyi, P. S., Pryor, E. E. J., Sarver, J. L., Cafiso, D. S., & Bushweller, J. H. (2013). Structural and dynamic studies of the transcription factor ERG reveal DNA binding is allosterically autoinhibited. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 110(33), 13374-13379. doi:10.1073/pnas.1301726110

2012

Leach, B. I., Kuntimaddi, A., Schmidt, C. R., Cierpicki, T., Johnson, S. A., & Bushweller, J. H. (2013). Leukemia Fusion Target AF9 Is an Intrinsically Disordered Transcriptional Regulator that Recruits Multiple Partners via Coupled Folding and Binding. STRUCTURE, 21(1), 176-183. doi:10.1016/j.str.2012.11.011

BUSHWELLER JOHN H., GREMBECKA JOLANTA., ILLENDULA ANURADHA., & DIXON LAUREN. (2012). INHIBITORS OF INV(16) LEUKEMIA.

2011

Chen, H., Ji, F., Olman, V., Mobley, C. K., Liu, Y., Zhou, Y., . . . Xu, Y. (2011). Optimal Mutation Sites for PRE Data Collection and Membrane Protein Structure Prediction. STRUCTURE, 19(4), 484-495. doi:10.1016/j.str.2011.02.002

2010

Fruh, V., Zhou, Y., Chen, D., Loch, C., AB, E., Grinkova, Y. N., . . . Siegal, G. (2010). Application of Fragment-Based Drug Discovery to Membrane Proteins: Identification of Ligands of the Integral Membrane Enzyme DsbB. CHEMISTRY & BIOLOGY, 17(8), 881-891. doi:10.1016/j.chembiol.2010.06.011

Corpora, T., Roudaia, L., Oo, Z. M., Chen, W., Manuylova, E., Cai, X., . . . Bushweller, J. H. (2010). Structure of the AML1-ETO NHR3-KA(RIIα) Complex and Its Contribution to AML1-ETO Activity. JOURNAL OF MOLECULAR BIOLOGY, 402(3), 560-577. doi:10.1016/j.jmb.2010.08.007

Park, S., Osmers, U., Raman, G., Schwantes, R. H., Diaz, M. O., & Bushweller, J. H. (2010). The PHD3 Domain of MLL Acts as a CYP33-Regulated Switch between MLL-Mediated Activation and Repression. BIOCHEMISTRY, 49(31), 6576-6586. doi:10.1021/bi1009387

Kamikubo, Y., Zhao, L., Wunderlich, M., Corpora, T., Hyde, R. K., Paul, T. A., . . . Liu, P. P. (2010). Accelerated Leukemogenesis by Truncated CBFβ-SMMHC Defective in High-Affinity Binding with RUNX1. CANCER CELL, 17(5), 455-468. doi:10.1016/j.ccr.2010.03.022