David J Shapiro

djshapir@illinois.edu

314d Roger Adams Lab
Office: (217) 333-1788
Lab: (217) 244-0207
Fax: (217) 244-5858

Mail to: Department of Biochemistry
419 Roger Adams Lab.
University of Illinois
600 S. Mathews Avenue
Urbana, IL 61801
Lab Page

David J Shapiro

Professor of Biochemistry

Research Topics

Drug Discovery, Endocrinology, Protein-Nucleic Acid Interactions, Receptor Biochemistry, Regulation of Gene Expression, Signal Transduction

Education

B.S. 1967 Brooklyn College
Ph.D. 1972 Purdue University
Postdoc. 1972-1973 Stanford University Medical School, 1973-1974 Stanford University
1985 - 1986 Guggenheim Fellow, Center for Cancer Research MIT

Identification and use of small molecule biomodulators as novel anticancer drugs and probes for new regulatory pathways

A novel strategy for identifying new regulatory strategies and anticancer drugs

Our approach is a novel way to do molecular biology in which unbiased high throughput screening is used to “interrogate the cell” to identify new pathways. We screen to develop a large database, and use simple follow-on assays and bioinformatics to identify small molecule biomodulators likely to exhibit a previously undescribed mode of action. Using this approach we identified an exceptionally promising first-in-class preclinical anticancer drug and a previously undescribed conserved pathway of hormone action.

A new essential action of hormones: Anticipatory activation of the Unfolded Protein Response (UPR)

The endoplasmic reticulum (EnR) stress sensor, the unfolded protein response (UPR), was known as a pathway that reacts to diverse forms of stress by activating signaling pathways that reduce protein production and increase protein-folding capacity. From our novel screening strategy, we showed that the steroid hormone estrogen bound to estrogen receptor (ER) very rapidly activates a different kind of UPR pathway that is activated in the absence of cell stress and anticipates a future need for increased protein folding capacity. Demonstrating relevance in humans, analysis of bioinformatic data from approximately 1,000 ER positive breast cancers shows that elevated expression of a UPR gene signature at diagnosis is a powerful new prognostic marker tightly correlated with subsequent tumor recurrence, therapy resistance and poor survival. This newly unveiled pathway of hormone action is conserved from insects to humans and between steroid hormones and peptide hormones. An important focus of our research is to understand how the very rapid hormone activation of the anticipatory UPR pathway couples to and authorizes subsequent actions of estrogen, the oncogenic hormone, epidermal growth factor (EGF), and other hormones, and how activation of this pathway protects cancer cells.

CRISPR-generated ER mutations from metastatic breast cancer

Estrogen, acting via ER, drives proliferation of more than 70% of human breast cancers. Recent studies show that therapies that target estrogen and ER often select for ER mutations D538G and Y537S. Using our modifications of CRISPR-Cas9 genome editing protocols, we isolated multiple breast cancer cell lines in which both copies of the wild type ER gene were replaced with ERD538G or ERY537S. This was perhaps the first example of using CRISPR to replace both copies of a wild type gene with a mutant gene in a cancer cell. In our biologically relevant anchorage independent 3-dimensional culture system, the ERD538G and ERY537S cells exhibit robust proliferation without estrogen and are resistant to current therapies. The molecular and biochemical basis for estrogen-independent, drug-resistant, growth of these cancer cells is a focus of current research.

UPR hyperactivation as a novel cancer targeting strategy

Cancer therapies that target estrogen production or binding to ER in breast cancer often select for resistance. Although many ovarian cancers contain ER, these highly lethal tumors do not respond to therapies targeting ER. Using our novel screening and drug development approach, we identified BHPI, a potent first-in-class non-competitive ER biomodulator that kills therapy-resistant ER positive cancer cells in cell culture and induces rapid and dramatic regression of breast cancers in mouse models. Notably, BHPI kills the ERY537S and ERD538G cells that are resistant to current therapies. BHPI is so effective because it works by hyperactivating the anticipatory UPR pathway described above, converting it from protective to toxic. Ovarian cancers are so lethal because they develop resistance to chemotherapy, often through overexpression of hard-to-target drug efflux pumps. Its novel mode of action enables BHPI to reverse multidrug resistance in ER positive ovarian cancer and reduce tumor mass and serum markers to undetectable levels. With our strong team of collaborators we are working to move BHPI forward toward clinical trials.

For background and more details see our Web site at www.life.uiuc.edu/shapiro . This site is accessible through the Lab Page link and in the Department of Biochemistry Web site.

Awards

2016 – Present Eugene Howe Scholar in Biochemistry

Representative Publications

Andruska, N., Zheng, X., Yang, X., Mao, C., Cherian, M., Mahapatra, L., Helferich, W.G. and Shapiro, D.J. (2015) An Estrogen Receptor Inhibitor Activates the Unfolded Protein Response, Blocks Protein Synthesis and Induces Tumor Regression. Proc. Natl. Acad. Sci. USA, 112(15) 4737-4742 (highlighted paper)

Andruska, N., Zheng, X., Yang, X., Helferich, W.G. and Shapiro, D.J. (2015) Anticipatory Estrogen Activation of the Unfolded Protein Response is Linked to Cell Proliferation and Poor Survival in Estrogen Receptor Positive Breast Cancer. Oncogene, 34(29):3760-3769. PMCID: PMC4377305

Zheng, X., Andruska, N., Lambrecht, M., He, S., Parissenti, A., Hergenrother, P., Nelson, E.R. and Shapiro, D.J. (2016) Targeting Multidrug Resistant Ovarian Cancer Through Estrogen Receptor Dependent Hyperactivation of the UPR. Oncotarget, OnLine July 24; doi: 10.18632/oncotarget.10819 (Dept. of Biochemistry, Colin Wraight Award. Outstanding graduate student paper of 2016)

Mao, C., Livezey, M., Kim, J.E., and Shapiro, D.J. (2016) CRISPR-Cas9-generated Cell Lines Expressing Estrogen Receptor Mutations from Metastatic Breast Cancer are Antiestrogen Resistant but are Killed by BHPI. Scientific Reports, OnLine 10/7/2016, doi: 10.1038/srep34753.

Yu, L., Andruska, N., Zheng, X. and Shapiro, D.J. (2016) Anticipatory Activation of the Unfolded Protein Response by Epidermal Growth Factor is Required for Immediate Early Gene Expression and Cell Proliferation. Mol. Cell. Endocrinol., 422:31-41. PMCID: PMC4919024

Shapiro D.J., Livezey, M., Yu, L., Zheng, X. and Andruska, N. (2016) Anticipatory UPR Activation: A Protective Pathway and Target in Cancer. Trends in Endocrinology and Metabolism (Cell Press), 27(10) 731-741. PMCID: PMC5035594

Zheng, X., Andruska, N., Yu, L., Mao, C., Kim, J. E., Livezey, M., Helferich, W. G., and Shapiro, D. J. (2016) Interplay between steroid hormone activation of the unfolded protein response and nuclear receptor action. Steroids, 114:2-6. PMCID: PMC5035163