RESEARCH FELLOWS

The UW Dermatology Division has been awarded a T32 Training Grant (2009-2014) from the NIH, “Training in Investigative Dermatology.” The mission of this grant is to train the next generation of independent investigators in dermatology and skin biology.

More information about research is available in the Dermatology Research Handbook.

The following trainees have been funded by this grant for their research on either a Skin Biology
Track
or Clinical Investigator Track.

Current Research Fellows
Past Research Fellows
 

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Current Research Fellows

hertz

Angie Hertz (Marson), PhD (10/1/2012 – Present)
Mentor: Steven F. Ziegler, PhD
Steven Ziegler Lab

Dr. Hertz hypothesizes that MMP10 plays a role in moderating Th1 immune reactions in the skin
through control of macrophage polarization. These M2 polarized macrophages are subsequently
able to polarized macrophages are subsequently able to active TREG cells and suppress the Th1 immune response. Therefore, Mmp10–/– mice should exhibit increasedTh1-­‐mediated inflammation as a result of decreased activation of suppressive TREG cells in a contact hypersensitivity model of skin inflammation. Dr. Hertz will use the well-­‐established DNCB–based CHS model in wild-­‐type and Mmp10–/– mice. In this model, which is established in the Parks lab, 6-­‐10 week old mice are sensitized to DNCB by a single application of DNCB on the abdomen. Five days later, the mice are challenged with a second application of DNCB on the back. Tissue sections will be collected from sites of both sensitization and challenge for histology and RNA analysis. The inguinal and axillary skin-­‐draining lymph nodes, as well as the spleen, will be harvested to isolate leukocytes for culture or flow cytometry analysis. Dr. Hertz expects to observe differences in the degree of inflammation in the histological sections, with an increase in macrophages and other lymphocytes. The skin-­‐draining lymph nodes reflect the cutaneous immune environment (Tomura et al. 2008), and the lab expects observe changes in the types of TREG cells present in the draining lymph nodes.

Publications

Hertz AL and Beavo JA. (2011) Cyclic Nucleotides and PDEs in monocyte differentiation. Francis SH, Conti M, Houslay MD (Eds). Handb Exp Pharmacol. 204: 365-90.

Hertz AL, Bender AT, Smith KC, Gilchrist M, Amieux PS, Aderem A, Beavo JA. (2009) Elevated cyclic AMP and PDE4 inhibition induce chemokine expression in human monocyte-derived macrophages. Proc Natl Acad Sci U S A. 106 (51): 21978-83.

Lyssand JS, DeFino MC, Tang XB, Hertz AL, Feller DB, Wacker JL, Adams ME, Hague
C. (2008) Blood pressure is regulated by an alpha1D-adrenergic receptor/dystrophin signalosome. J Biol Chem. 283(27):18792-18800.
 

Marson A, Lawn RM, Mikita T. (2005) Oxidized Low Density Lipoprotein Blocks Lipopolysaccharide-Induced Interferon β Synthesis in Human Macrophages by
Interfering with IRF3 Activation. J Biol Chem.  279: 28781-28788.

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Clay Sontheimer, MD (7/2013-6/2014)
Mentors: Keith Elkon, MD

Elkon lab

Innate immunity in the skin following ultraviolet-B exposure and the mechanisms of
photosensitivity in lupus


Thomas Stoklasek, PhD (7/2013-6/2014)
Mentors: Stephen Zeigler, PhD

Zeigler lab

The Role of Innate Lymphoid Cells in Systemic Sclerosis

 

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Soyeon Lippman, PhD (9/1/2010-10/1/2012)
Mentors: Pradeep Singh, MD & Colin  PhD

Manoil lab
The purpose of Dr. Lippman’s project is to identify cellular functions of Pseudomonas aeruginosa that could be targeted to inhibit the establishment and maintenance of wound infections. A recently developed genome-wide genetic approach (Tn-seq) will be employed to identify P. aeruginosa genes that are required for wound infections and those genes that are required for persistence of the infection even under antimicrobial treatments. To this end, a genome-scale mutant library has been generated encompassing approximately forty individual mutations per gene in P. aeruginosa. Preliminary studies have uncovered an uncharacterized hypothetical membrane protein, PA5528, to be significantly attenuated in its ability to establish an infection in a mouse wound model. Intriguingly, bacterial cells with an inactivated PA5528 gene product also display highly increased sensitivity to the aminoglycoside antibiotic, tobramycin. Combination of powerful global mutant analysis and mouse wound model is expected to yield discovery of novel drug targets for treatment of wound infections. Dr. Lippman was awarded a  NIH F32 grant for continued research. She is currently studying in the School of Global Health at the University of washington.

Presentations
Gordon Conference on Drug Resistance, Easton, MA (2012). Discovering bacterial drug targets that avoid antibiotic resistance. Lippman SI, Singh P, Olerud J, Manoil C.

University of Washington, Dept of Genome Sciences, Seattle, WA (2012). Identifying Bacterial
Functions Required for Skin Infection While Saving 5000 Mice. Lippman SI, Singh P, Olerud J, Manoil C.

Systems Biology and the Microbiome Symposium, Institute for Systems Biology, Seattle, WA (2012). Genome-wide Screen for Bacterial Functions Required for Skin Infection. Lippman SI, Singh P,
Olerud J, Manoil C.

University of Washington, Dept of Dermatology, Seattle, WA (2011). Treating Chronic Wound Infections: Finding New Drug Targets of P. aeruginosa Using Genome-wide Tools. Lippman SI, Singh P, Manoil C.

Fred Hutchinson Cancer Research Center, Seattle, WA (2010). How Do Yeast Cells Coordinate
Growth With Nutrient Availability? Lippman SI and Broach JR.

University of Washington, Dept of Genome Sciences, Seattle, WA (2010). Integration of Multiple
External Cues to Regulate Growth in the Budding Yeast. Lippman SI and Broach JR.

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Past Research Fellows

Maryam Ganjehzadeh Rohani (10/1/09-9/30/10)
Mentor: William Parks, PhD

Dr. Rohani worked under the supervision of Dr. Bill Parks, studying the signaling pathways involved in skin re-epithelialization. During wound injury keratinocytes in contact with collagen type I, the ligand for a2b1 integrin, induce collagenase-I (MMP1). Cells expressing MMP1 have migratory phenotype, which suggest involvement of Rho GTPase signaling pathway. Investigating the role of Rho GTPases (Cdc42, RhoA and Rac1) in regulation of MMP1 revealed that Cdc42 has an inhibitory effect, but RhoA induces MMP1 expression. Furthermore, as the previous studies had shown that ERK MAP kinase signaling is necessary for induction of MMP1, Dr.Rohani tested the cross talk between ERK and Rho GTPases and found that they act independently. The in situ Rho GTP affinity assay also suggests an increase in activity of Rho at the wound edges, which is in agreement with the role of RhoA in induction of MMP1.

Dr. Rohani was awarded a NIH NRSA F32 Postdoctoral Fellowship Award in September 2010. In 2014 Maryam accepted a position at Cedars-Sinai, Los Angeles.

Publications
Maryam G. Rohani, Dennis H DiJulio, Jonathan Y An, Beth M Hacker, Beverly A Dale and Whasun O Chung. “PAR1- and PAR2-induced innate immune markers are negatively regulated by PI3K/Akt signaling pathway in oral keratinocytes”, BMC Immunology 2010, 11:53 PMID: 21029417

Maryam G. Rohani,
Peter Chen, William C. Parks “Rac-1 Modulates Expression of MMP-1 in Keratiocytes in Contact with Type I Collagen, Biennial Meeting of the American Society for Matrix
Biology, Charleston, SC, October 2010
Whasun O. Chung, Henrik Dommisch and Maryam G. Rohani, Pattern Recognition Receptors of Oral Epithelia in “Perspectives on Pattern Recognition Receptors”, Monica D. Fournier, Nova publisher, 2011

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Travis Biechele, PhD (6/30/10-6/30/12)
Mentors: Randall Moon, PhD and Andy Chien, MD, PhD

Moon Lab
As the Wnt/ß-catenin signaling pathway is linked to melanoma pathogenesis and to patient survival,
Dr. Biechele and his mentors, Drs. Randy Moon and Andy Chien, conducted a kinome siRNA screen in melanoma cells to expand their understanding of kinases that regulate this pathway, and to illuminate potential therapeutic directions. They found that BRAF/MAPK signaling, which is constitutively activated in many melanomas by the BRAFV600E mutation, negatively regulates Wnt/ß-catenin signaling in human melanoma cells. As inhibitors of BRAFV600E show promise in ongoing clinical trials we investigated whether altering Wnt/ß-catenin signaling might enhance the efficacy of a BRAFV600E inhibitor. Surprisingly, endogenous ß-catenin is required for the BRAFV600E inhibitor to induce apoptosis in melanoma, while activation of Wnt/ß-catenin signaling strongly synergizes with the BRAFV600E inhibitor to decrease tumor growth in vivo and to increase apoptosis in vitro. This synergistic enhancement of apoptosis correlates with a reduction in the steady-state levels of a ß-catenin antagonist, AXIN1. In support of the hypothesis that AXIN1 is a mediator rather than marker
of apoptosis, melanoma cell lines that are resistant to apoptosis after treatment with a BRAFV600E inhibitor become susceptible, and undergo apoptosis, when levels of AXIN1/2 are reduced by siRNA. These findings point to a significant role for Wnt/ß-catenin signaling and AXIN1 in regulating the efficacy of inhibitors of BRAFV600E, and lay a novel foundation for developing potential combination therapies and biomarkers.

In July 2012 Dr. Biechele joined the team at Seattle Genetics to continue his research.

Publications & Presentations

Travis L. Biechele, Rima M. Kulikauskas, Olivia M. Lucero, Rachel A. Toroni, Reyna D. Swift, Richard
G. James, Nick C. Robin, David W. Dawson, Randall T. Moon, and Andy J. Chien. Wnt/ß-catenin signaling and AXIN1 regulate apoptosis mediated by inhibition of BRAFV600E kinase in human melanoma. Science Signaling

December 7, 2011. Works-in-Progress presentation to University of Washington Division of Dermatology Grand Rounds.

June 29-July 3, 2011. Presentation to Wnt Meeting 2011, Los Angeles, CA. Abstract submitted.

September 9, 2010. Works-in-progress presentation to University of Washington Division of Dermatology Grand Rounds.

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zhao

Ge (Alice) Zhao MD/PhD (6/30/10- 6/23/11)
Mentors: John Olerud, MD and Philip Fleckman, MD

Alice worked with Dr. John Olerud developing biofilm challenged chronic wounds in diabetic mice. Chronic wounds are a major clinical problem and research is limited due to lack of a reproducible animal model. Previous work has shown that transferring Pseudomonas biofilm to wounds on
diabetic mice significantly delays wound healing. During the previous year, we have optimized wound management, studied wound healing process at extended time course, evaluated oxygen distribution
in biofilm, and characterized gene expression in mice tissue in response to biofilm challenge. The results showed that this is a reproducible delayed wound model, providing at least a two week window to analyze unhealed wounds, and sharing similar host wound features as in human chronic wounds. We have also collaborated with faculty in the Department of Microbiology (Dr. Pradeep Singh and Dr. Joseph Mougous) to extend the application of this model in studying biofilm in vivo and in studying different pathogens (S. aureus and Acinetobacter). Another research area Dr. Zhao evaluated was anti-infection effect of percutaneous devices. Dr. Zhao and her mentors found that integration of porous polymer material with skin can significantly reduce bacterial invasion in the implants. The results were presented at the Wound Healing Society Annual Meeting (Apr 14-17, 2011) and Society for Investigative Dermatology Annual Meeting (May 4-7, 2011).

Dr. Zhao began a Medicine Internship in June 2011 and entered Dermatology Residency in July 2012 at the University of Washington.

Publications & Presentations

Zhao G, Hochwalt PC, Usui ML, Underwood RA, Singh PK, James GA, Stewart PS, Fleckman P, Olerud JE. Delayed wound healing in diabetic (db/db) mice with Pseudomonas aeruginosa biofilm challenge: a model for the study of chronic wounds. Wound Repair Regen.2010 Aug; Epub ahead of print(0): 0. Cited in PubMed; PMID: 20731798.

Young Investigator Award from the Center for Biofilm Engineering, 2010
Anita Roberts Honorable Mention Award from the Wound Healing Society, 2010
Best Speaker Award from University of Washington Postdoctoral Symposium, 2010.

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Jessica White, PhD (7/1/10-1/31/12)
Mentor: David Koelle, MD

Almost every known protein-coding ORF of KSHV were PCR-cloned into a flexible “entry” vector by the Haas, Sturzl, Hayward, or Schulz labs and were provided to Dr. White and her mentor, David Koelle, for this research project. Dr. White has completed PCR-cloning the few KSHV ORFs missing from these plasmid collections in to the entry vector, using KSHV strain BCBL-1 DNA or BC-1 DNA. All of the KSHV ORFs have been sub-cloned into the CD4-style custom destination vector and over half have been
sub-cloned the CD8-style expression vector. She has begun expressing KSHV protein by adding these KSHV clones to commercially available E. coli transcription/translation lysate. Dot blots have been performed using the 6-His tags present for verification of protein expression. Cryopreserved PBMC isolated from anticoagulated venous blood collected from 20 HIV-negative, KSHV-infected
(serologically confirmed) persons from the Seattle area have been completed. Serum samples have been separated and further antibody testing is being performed in Dr. Vieira’s lab. Vero cells constitutively infected with recombinant HHV-8 (rKSHV.219) expressing eGFP, created by Dr. Vieira,
will be used for use in enrichment of KSHV-specific CD8 T-cells. Dr. White has optimized the induction lytic viral replication to ensure presentation of both lytic and latent viral proteins to CD8 T-cells; completed flow cytometry training; and developed an ICC protocol to sort for live/CD3/CD8/CD137 (recently activated CD8 T-cells) and the detection of IL-2, TNFα, and IFN-γ cytokines. This will allow for the detection of KSHV-specific memory CD8 T-cells after exposure to dendritic cells presenting apoptotic rKSHV.219 infected vero cell debris antigen. We predict that the CD137-high cells but not the CD137-low cells will be KSHV-specific. To check this, autologous B-cells, immortalized by EBV a convenient way of propagating “self” cells for immunology research, will be infected with recombinant KSHV. The bulk-expanded T-cells (CD137-high or control CD137-low) will be added and their activation detected using ICC. Dr. White has generated autologous B-cells immortalized by EBV for all Seattle study participants collected. Applied for F32 funding in January 2011. Application title: Comprehensive measurement of KSHV-specific CD4 and CD8 T-cell responses in humans

In February 2012 Dr. White joined the Vaccine Adjuvant Formulation Development Group as a Sr. Program Associate, Technology Solutions at PATH. She continues to receive training and gain experience in formulation research and vaccine adjuvant development on a project funded by the Bill and Melinda Gates Foundation, working under Dr. Dexiang Chen.

Presentations
June 2012. Works-in-Progress lecture to University of Washington, Division of Dermatology Grand Rounds.

November 2010. Works-in-Progress presentation to University of Washington, Division of Dermatology
Grand Rounds.

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esparza

Ed Esparza, MD, PhD (7/1/10-6/30/11)
Mentor: David Rawlings, MD

Rawlings lab
Cutaneous Tcell lymphoma (CTCL) is a skin cancer that can spread to other organs and cause significant morbidity and mortality in advanced stages. The clinical and histologic features of CTCL overlap with benign inflammatory skin conditions that creates diagnostic and treatment dilemmas for patients and treating physicians. The goal of Dr. Esparza’s project has been to develop novel
molecular markers of disease, such as altered expression of tumor suppressors like A20 and
aberrant activation of oncogenic pathways like the NF‑kB cascade, which will aid our ability to
accurately distinguish CTCL from non-neoplastic inflammatory conditions and possibly prognosticate the disease course for a given patient. Dr. Esparza was awarded a mentorship award that facilitated collaboration with Drs. Anne Lind and Dongsi Lu at Washington University in dermatopathology. There, he was able to develop novel immunohistochemical tools on formalin-fixed skin biopsies to analyze A20 expression and NF‑kB activation in cutaneous lymphoproliferative disorders. Ongoing characterization of distinctive A20 expression and NF‑kB activation may distinguish between CTCL and benign mimickers. In addition, these distinctive signatures may eventually allow for tailoring therapies if a given condition is dependent on or independent of NF‑kB activation in transforming to lymphoma. Complementing these translational techniques, Dr. Esparza has been characterizing how A20 regulates NF‑kB activation using CTCL cell lines in the laboratory of David Rawlings. Although A20 is expressed in these cell lines, they have observed mutations in the coding sequence of A20 that may lead to functionally inactive protein. They are now characterizing the relevance of these mutations to A20 function in regulating NF‑kB activation and lymphomagenesis. Over the upcoming year, the results from these complementary approaches will be presented as an abstract at the American Society of Dermatopathology meeting on October 2011 and likely a manuscript submission.

In July 2011, Dr. Esparza joined the Dermatology practice at Virginia Mason Medical Center, Seattle.

DermPath Mentorship Award, September 2010 Studied with Drs. Anne Lind and Dongsi Lu at Washington University Department of Dermatopathology.

 

 

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