Amanda C. Poholek, Ph.D.

  • Director of the HSSC@CHP NextGen Sequencing Core Facility
  • Assistant Professor of Pediatrics 

Dr. Poholek did her undergraduate training at Fordham University in Bronx, NY and received her Ph.D. from Yale University in New Haven CT. At Yale, she trained with Dr. Joe Craft where she began her interest in immune-mediated disease and how transcription factors contribute to T cell differentiation and function. To further understand how transcription factors contribute to differentiation at the chromatin level, she did her post-doctoral training at the NIH with Dr. John O’Shea. There she learned how NextGen Sequencing can be used to interrogate chromatin accessibility, epigenetics and transcriptome analysis. She moved to the University of Pittsburgh in 2015 to begin her own lab and became Director of the HSSC@CHP NextGen Sequencing Core Facility in 2017. 

Professional Affiliations/Society Memberships

  • American Association of Immunology

Major Lectureships and Seminars

  • “IL-10 Induces an Autoregulatory Loop in T Cells that Promotes Blimp-1 Restriction of Cell Expansion,” platform presentation, Cold Spring Harbor Laboratory Meeting: Gene Expression and Signaling in the Immune System, Cold Spring Harbor, N.Y., April 2016 

Representative Publications

Poholek AC, Jankovic D, Villarino AV, Petermann F, Hettinga A, Shouval DS, Snapper SB, Kaech SM, Brooks SR, Vahedi G, Sher A, Kanno Y, O'Shea JJ. IL-10 induces a STAT3-dependent autoregulatory loop in Th2 cells that promotes Blimp-1 restriction of cell expansion via antagonism of STAT5 target genes. Science Immunology. Nov 11, Vol.1 Issue 5. 2016. http://immunology.sciencemag.org/content/1/5/eaaf8612.full

Shih HY, Sciume G, Poholek AC, Vahedi G, Hirahara K, Villarino A, Bonelli M, Bosselut R, Kanno Y, Muljo S, O’Shea JJ. Transcriptional and epigenetic networks of helper T cells and innate lymphoid cells. Immunological Reviews. Sept;261(1):23-49. 2014. http://onlinelibrary.wiley.com/doi/10.1111/imr.12208/abstract

Nakayamada S*, Poholek AC*, Lu KT, Takahashi H, Hirahara K, Kato, M, Iwata S, Cannons JL, Schwartzberg PL, Vahedi G, Sun H, Kanno Y, O’Shea JJ. Type I Interferon induces binding of STAT1 to Bcl6: Divergent Roles of STAT-family transcription factors in the TFH cell genetic program. The Journal of Immunology. Mar 1;192(5):2156-66. 2014. http://www.jimmunol.org/content/192/5/2156.long

Vahedi G*, Poholek AC*, Hand TW, Laurence A, Kanno Y, O’Shea JJ, Hirahara K. Helper T Cell Identity and Evolution of Differential Transcriptomes and Epigenomes. Immunological Reviews. Mar;252(1):24-40. 2013. http://onlinelibrary.wiley.com/doi/10.1111/imr.12037/abstract

Poholek AC, Hansen K, Hernandez S, Eto D, Chandele A, Weinstein JS, Dong X, Odegard J, Kaech SM, Dent AL, Crotty S, Craft J. In vivo regulation of Bcl6 and T follicular helper cell development. The Journal of Immunology, 185(1):313-26, 2010. http://www.jimmunol.org/content/185/1/313.long

Johnston R*, Poholek AC*, DiToro D, Yusuf I, Eto D, Barnett B, Dent A, Craft J, and Crotty S. Bcl6 and Blimp-1 are reciprocal and antagonist regulators of T follicular helper cell differentiation. Science. 325(5943):1006-10, 2009 http://science.sciencemag.org/content/325/5943/1006.long

Research Interests

The Poholek Lab is interested in the molecular and epigenetic mechanisms of transcription factor control of T cell differentiation and function. Our goal is to understand how T cells see their environment and translate those signals through epigenetic and transcriptomic changes into unique cellular functions. This is important because T cell dysfunction underlies many human diseases and rationale vaccine design requires an understanding of how T cells function in different contexts. Currently we are interested in how Blimp-1, a transcriptional repressor present in many cell types, functions uniquely in T cells in the context of normal function and in disease. Blimp-1 is associated with constraining T-cell-mediated autoimmune disease and is highly expressed in exhausted T cells present in chronic viral infection and the tumor microenvironment. Our team is working to understand the factors that regulate expression of Blimp-1 in CD4 and CD8 T cells, and to identify the non-coding genetic regulatory elements such as enhancers that are critical for cell-type-specific expression of Blimp-1 in order to understand how Blimp-1 functions in asthma, tumors, and homeostatic function of several tissues.

 

To understand how transcription factors control immune function, we take a multi-disciplinary approach, combining both in vitro and in vivo model systems using several complex genetic tools. Technically, we use multicolor flow cytometry as a single cell quantitative assay of protein expression and NextGen Sequencing technologies such as RNAseq, ChIPseq, and ATACseq to identify epigenomic and transcriptomic changes in T cells.

 

Current projects in the lab:

Identification of enhancers controlling cell-type dependent expression of Blimp-1

Using ChIP-seq to identify enhancers and a genetic reporter of Blimp-1 expression, we are exploring the factors that are required to drive Blimp-1 expression and determining which non-coding enhancers are relevant to control expression of Blimp-1 in different cell types.

 

Determining cell-type specific function of Blimp-1

Using RNA-seq and ChIP-seq, we are exploring how Blimp-1 functions in different immune cell types. Genetic tools allow us to delete Blimp-1 in specific cell types (constitutively or inducibly) or overexpress Blimp-1 in a dose-dependent cell type specific manner.

 

Blimp-1 mediated Th2 cell development in the lung impacting allergic airway inflammation

Using a murine model of allergic asthma, we are exploring the requirement of Blimp-1 in asthma disease.

 

Epigenetic regulation of tumor-infiltrating T cells

Using a new low-cell number ChIPseq assay (CUT&RUN) we are exploring the epigenetics of T cells in murine tumors in collaboration with Dr. Greg Delgoffe at the Hillman Cancer Center.

Division

Center / Institute

Lab