Jan Peters PhD May 2015 CV

J A N P E T E R S , P H D7153 Tulip Trail Dr. ~ Memphis, TN 38133

[email protected] 443.416.8202

Permanent Resident (Green Card) www.linkedin.com/pub/jan-peters/8/a1/908

PR O F I L E

Innovative, results-driven, and fellowship-trained preclinical research scientist and principal investigator with experience in both commercially funded and grant funded research. Background developing and managing experimental assays using in vitro and in vivo techniques in cell/tissue culture models / live animal models. Extensive publication history in peer-reviewed publications (numerous citations) and experience presenting to groups and collaborating on writing (papers, grant-writing, SOPs, etc.).

AREAS OF STRENGTH – Biochemistry, Molecular Biology (DNA- & RNA-based methods), Cell Biology, Computational Biology, Microbiology, Immunology.

ED U C A T I O N

LEIBNIZ UNIVERSITY HANNOVER / MEDICAL SCHOOL HANNOVER, Hannover, Germany 2001-2004PhD – Biochemistry & Microbiology (Doctor of Science, Dr. rer. nat.) Doctoral Thesis – “Host Cell Responses to Intracellular Bacteria: Chlamydial Persistence and Comparative Analysis of Salmonella Typhimurium to Chlamydia during Productive Infection”

LEIBNIZ UNIVERSITY HANNOVER / MEDICAL SCHOOL HANNOVER, Hannover, Germany 1998-2001Diploma – Biochemistry (equivalent to Master of Science / MSc)Master’s Thesis – “Examination of Transcription and Expression of Chlamydial Genes”

LEIBNIZ UNIVERSITY HANNOVER / MEDICAL SCHOOL HANNOVER, Hannover, Germany 1996-1998Pre-Diploma – Biochemistry (equivalent to Bachelor of Science / BSc)

FE L L O W S H I P S

THE UNIVERSITY OF TENNESSEE HEALTH SCIENCE CENTER (UTHSC), Memphis, TN 2008-2011DEPARTMENT OF MICROBIOLOGY, IMMUNOLOGY AND BIOCHEMISTRY Postdoctoral Fellowship – Chlamydia: IFN-, Protein Characterization, Lipids, sRNA AnalysisPostdoctoral Fellowship – Virology / Immunology

UNIVERSITY OF MARYLAND – DEPARTMENT OF MICROBIAL PATHOGENESIS, Baltimore, MD 2005-2008Postdoctoral Fellowship – Chlamydia: Type III Secretion Systems & Effector Proteins

AD D I T I O N A L TR A I N I N G

BioPharma Institute – GMP0: Documentation and Recordkeeping Department of Justice – Select Agents Clearance The Institute for Genomic Research (TIGR) – Prokaryotic Genome Annotation & Analysis Improvision (Germany) – Cell Imaging Workshop ABI – ABI PISM® 7000 Real-Time RT-PCR TrainingEuropean Molecular Biology Organization (EMBO) – DNA Microarrays: Applications and Data Analysis

JAN PETERS PRECLINICAL RESEARCH PAGE 2 OF 8

RE S E A R C H EX P E R I E N C E

THE UNIVERSITY OF TENNESSEE HEALTH SCIENCE CENTER (UTHSC), Memphis, TN 2011-PresentDEPARTMENT OF MICROBIOLOGY, IMMUNOLOGY AND BIOCHEMISTRY

Research Assistant Professor Conduct preclinical research projects funded by grant from Center for Integrated and Translational Genomics. Coordinate with multidisciplinary research teams involved in related research; supervise junior researchers, interns, and technician and work in collaboration with research partners. Oversee data interpretation, reporting, and presentation / publication of research reports – both internally, in multidisciplinary team environment, and externally in international peer-reviewed journals. Write protocols for Institutional Biosafety (IBC) and Animal Care and Use (IACUC) Committees. Conduct and coordinate collaborative grant writing of grant applications for external funding. Teach Molecular Biology classes in Microbial Pathogenesis seminar for graduate students.

Published results of inhibitor studies with 4-phenyl imidazole (4-PI) in cell culture model

o Identified Acyl-CoA:cholesterol acyltransferase (ACAT) as potential target protein for 4-PIo Demonstrated dependency of C.trachomatis growth on ACAT activity in cell culture model

Handled functional analysis of chlamydial protein CT149 by biochemical assays Used bioinformatics tools to model structure of all proteins of 4 distinct chlamydial species Prepared fluorescent Chlamydia for in vivo and in vitro live imaging

o Cloned of fluorescent marker into Chlamydia muridarum plasmid o Transformed construct into C. muridarum

Developed independent research programs:

o Computational biology approach for functional analysis of chlamydial proteins using bioinformatics tools (e.g. prediction of protein structure, ligand and substrate binding, Autodocking, protein localization)

o Role of lipids in immunity during Chlamydia infection

REGIONAL BIOCONTAINMENT LABORATORY, Memphis TN 2011-present

Scientific CoordinatorDevelop and review standard operation procedures (SOPs) for BSL-3 work. Conduct experimental assay design, testing, and validation, and managed compliance with industry and institutional standards. Supervise technician and develop assays for Janus liquid handling system together with technician. Handle or schedule calibration and maintenance of sensitive BSL-2 and BSL-3 laboratory equipment – AriaII flow cytometer, Luminex reader, Biorad qPCR, Janus robotic liquid transfer system, EnVision multilabel plate reader, BioTek plate reader, DeltaVision fluorescence microscope, Perkin Elmer IVIS Spectrum live imager.

Worked with multiple investigators to develop experiments and projects with RBL-housed equipment and handle troubleshooting of analysis results.

Trained investigators on in vitro work under BSL-3 safety conditions in compliance with SOPs. Aided in RBL-housed studies:

o Isolation of Chlamydia from wild mice tissue (in progress)o In vivo drug study against Chlamydia psittaci in mice (finished)

Executed Luminex and qPCR assays for investigators


THE UNIVERSITY OF TENNESSEE HEALTH SCIENCE CENTER (UTHSC), Memphis, TN 2008-2011DEPARTMENT OF MICROBIOLOGY, IMMUNOLOGY AND BIOCHEMISTRY

Postdoctoral Fellow – Chlamydia: IFN-, Protein Characterization, Lipids, sRNA Functional Analysis 2009-2011Advisors: Gerald Byrne, PhD and Robert Belland, PhDWorking independently, managed, planned and conducted experiments within projects. Mentored rotation students in research. Worked closely with investigators from commercial and academic research.

Published functional analysis of chlamydial protein CT149 o Used bioinformatics tools to analyze sequence / structural homologies and identified

potential signal N-terminal peptide o Demonstrated carboxylic esterase activity of CT149 in hydrolysis assay using HPLCo Showed expression of CT149 in HeLa cells results in decrease of cellular cholesteryl

esters in cell culture modelo Produced mono-specific antibody against CT149 in Balb / c mice for localization studies

Conducted functional analysis of chlamydial protein CT359o Demonstrated biotin transport activity of CT359 in E. coli surrogate system using self-

developed biochemical fluorescence based transport assay Conducted analysis of Chlamydia growth inhibition by inhibitor 4-PI in cell culture model

o Investigated potential eukaryotic protein targets by siRNA knockdown in cell culture system Tested regulatory effect of plasmid coded small RNAs (sRNAs) CHL1.1 and CHL1.2 on gene

expression in E. coli surrogate system o Identified prospective small protein coded within CHL1.1 using bioinformatics toolso Analyzed potential regulatory function of protein

Studied additional host cell processes after Interferon- treatment inducing chlamydial persistence in cell culture system

Analyzed riboswitch activity of chlamydial sRNAs using E. coli surrogate system / Northern blots

Postdoctoral Fellow – Virology / Immunology 2008-2009Advisor: Kui Li, M.D., PhDIn similar role, conducted experimental assay design, testing, and validation, and managed compliance with industry and institutional standards. Mentored rotation students in their research.

Investigated influence of Sphingosine-1 phosphate phosphatase 2 (SGPP2) on polyI:polyC and Sendai Virus induced IFN-β promoter activity in reporter gene assay and Western blot analysis

Created Tet-regulated Myc-SGPP2 expressing stable HeLa cell line Screened plasmid cDNA library for cytokines with Hepatitis C anti-viral properties using stable

transfected replicon cell line

UNIVERSITY OF MARYLAND – DEPARTMENT OF MICROBIAL PATHOGENESIS, Baltimore, MD 2005-2008

Postdoctoral Fellow – Chlamydia: Type III Secretion Systems & Effector ProteinsAdvisor: Patrik Bavoil, PhD

Analyzed binding of Chlamydia caviae GPIC Type III secretion system (T3SS) effector CopN to eukaryotic Aldolase using pull-down and co-immuno precipitation assays

Established fluorescence-based β-lactamase (-lac) secretion assay using -lac fusion proteins in rabbit EPECs and Salmonella Typhimurium to investigate translocation of putative chlamydial T3SS effector proteins

Created genomic β-lac fusion protein libraries of Chlamydia trachomatis serovar D and L2o Screened library for new chlamydial T3S effector proteinso Identified prospective new T3SS effector candidate from C. trachomatis L2


Facilitated microbiology case-based conference for dentistry studentsMEDICAL SCHOOL HANNOVER, Hannover, Germany 2000-2004DEPARTMENT OF MEDICAL MICROBIOLOGY AND HOSPITAL EPIDEMIOLOGY

PhD Researcher – Chlamydia: Persistence, Host Response, Protein Characterization) 2001-2004Advisor: Andreas Klos, MD

Established 3 models of persistence of C. trachomatis D and C. pneumoniae in in vitro cell culture system using HeLa and Hep-2 cells

Demonstrated differences in gene and protein expression patterns of HeLa cells during infection of C. pneumoniae in 3 different models of persistence compared to productive infection

o Demonstrated alteration of host gene expression in real-time quantitative PCR o Measured alteration of host protein expression in ELISA for important chemokines o Assessed importance of these differences for diseases caused by persisting Chlamydia

Analyzed gene expression of eukaryotic host cells after infection with Salmonella Typhimurium and evaluated these differences in terms of unique host cell responses to Chlamydia

Performed Affymetrix gene arrays of infected HeLa cells to analyze differences in host cell gene expression regulation between intracellular S. Typhimurium wt and SPI-2 T3SS mutant

Investigated cellular effects of C. trachomatis D putative toxin CT166 in HeLa compared to Clostridium difficile Toxin B and Salmonella protein SopE

o Showed phenotypic changes of cytoskeleton after CT166 expression in cell culture model Tested inhibitory effect of various antagonists of C3A-receptor in Ca2+-assay with FURA-2AM

Graduate Researcher – Chlamydia, Chlamydial Gene Regulation 2000-2001Advisor: Andreas Klos, MD

Developed gene expression assay for C3A receptor and chlamydial genes with Taqman probes and SYBR green (Taqman qPCR)

Managed cloning and expression of GFP-fusion proteins of selected chlamydial proteins in HeLa cells after transfection

SE L E C T E D TE C H N I Q U E S

Biochemistry Protein purification from bacteria, animal tissue and cell lines Protein separation by sodium-dodecylsulfate-polyacrylamidgel electrophorese (SDS-PAGE) Western blot analysis In-vitro enzyme activity assays Pull-down assay for protein-protein interaction Recombinant protein expression Lipid extraction Generation of fusion-proteins In-vitro translation High performance liquid chromatography (HPLC)

Molecular Biology DNA-based methods:

o Plasmid and genomic DNA purification from cell culture or tissue o Qualitative, quantitative, and preparative polymerase chain reaction (PCR) o Molecular cloning o Sequencing by Sanger dideoxynucleotide-method o Gel electrophorese mobility shift assay (GEMSA)


o Mutation of target genes by site-directed mutagenesis RNA-based methods:

o Purification of RNA o Reverse transcription (RT) o Conventional RT-PCR and real-time quantitative PCR (qPCR)o Gene expression analysis by Affymetrix gene array o Northern blotso In-vitro transcription

Cell Biology Culture of eukaryotic cells (cell lines and primary cells) Purification of immune-cells Transient and stable Transfection of eukaryotic cells Tetracyclin (Tet)-regulated protein expression in mammalian cells Reporter gene assays Intracellular Ca2+ measurement with FURA-2AM Cell fractionation

Computational Biology Utilization of bioinformatics tools for prediction of:

o Protein structure and structural homology to other proteinso Protein-protein interactiono Protein-ligand / substrate interactiono Protein localizationo Signal sequence predictiono sRNA identification and target prediction

Microbiology Culture and purification of bacteria Diagnostic of bacteria - Serotyping Infection of eukaryotic cells with Gram-negative bacteria

Immunology Enzyme linked immunosorbent assay (ELISA) Immuno-fluorescence assay (IFA) Co-immuno precipitation Flow cytometry basics / fluorescence-activated cell sorting (FACS) TUNEL assay T-cell activation

Microscopy Light microscopy & Electron microscopy Fluorescence microscopy (DeltaVision, confocal, deconvolution)

Software (selection) Web-based prediction software (e.g. SignalP, ExPAsSy tools, InterPro) Vector NTI and Geneious (DNA / protein database and manipulation software) GenMAPP (metabolic pathway software) & GenBank PyRx (Autodocking software) ClustalX and NJplot


Additional – ImageJ, Swiss-PDB viewer, Genome workbench (genome & DNA software), BioeditAW A R D S & HO N O R S

2010 1st Poster Prize in 3rd Annual UTHSC Postdoc Research Day for “Esterase Activity of Chlamydia trachomatis CT149 Gene Product Results in Cholesteryl Ester Hydrolysis When Expressed in HeLa Cells”

2007 AAAS Excellence in Science Award

PR O F E S S I O N A L A S S O C I A T I O N S

Ad Hoc Reviewer – Frontiers in Innate Immunity 2012-presentAmerican Association for Laboratory Animal Science (ALAAS) 2012-presentAdvancing Association for Advancement of Science (AAAS) 2008-presentAmerican Society for Microbiology 2005-presentPrevious: Chair – University of Maryland Biomedical Sciences Postdoc Network, St. Jude’s Institutional

Biosafety Committee (IBC), Society for Biochemistry & Molecular Biology

PU B L I C A T I O N S & PR E S E N T A T I O N S

PEER-REVIEWED MANUSCRIPTS:1. Peters J* and Byrne GI. Chlamydia trachomatis Growth Depends on Eukaryotic Cholesterol

Esterification And Is Affected by Acyl-CoA.Cholesterol Acyltransferase Inhibition, Pathog Dis, 2015 April (published, doi. 10.1093 / femspd / ftv028) *corresponding author

2. Cox JV, Abdelrahman YM, Peters J, Yao J, Naher N, Rock CO, and Belland RJ. Chlamydia trachomatis Utilizes Mammalian CLA1 Lipid Transporter to Acquire Host Phosphatidylcholine Essential for Growth, Cell Microbiol, 2015 Mar (submitted)

3. Abdelsamed H*, Peters J*, and Byrne GI. Genetic variation in Chlamydia trachomatis and their hosts: impact on disease severity and tissue tropism. Future Microbiol, 2013 Sept, 8 (9) 1129-1146; *equal contribution (11 citations)

4. Peters, J, Onguri, V, Nishimoto, SK, Marion TN, and Byrne, GI. Chlamydia trachomatis CT149 protein exhibits esterase activity in vitro and catalyzes cholesteryl ester hydrolysis when expressed in HeLa cells. Microbes and Infection, 2012 Aug, 14 (13) 1196-1204

5. Klos, A, Thalmann J, Peters J, Gerard H, Hudson A. transcript profile of persistent Chlamydophila (Chlamydia) pneumoniae in vitro depends on means by which persistence is induced. FEMS Microbiol Letters, 2009 Feb; 209 (1).120-126 (26 citations)

6. Peters J, Wilson DP, Myers G, Timms P, Bavoil PM. Type III Secretion à la Chlamydia. Trends Microbiol. 2007 Jun; 15(6).241-51. (137 citations)

7. Peters J, Hess S, Endlich K, Thalmann J, Holzberg D, Kracht M, Schaefer M, Bartling G, and Klos A. Silencing or Permanent Activation: Host-Cell Responses in Models of Persistent Chlamydia pneumoniae Infection. Cell Microbiol, 2005 Aug; 7(8).1099-108. (30 citations)

8. Hess S, Peters J, Bartling G, Rheinheimer C, Hegde P, Magid-Slav M, Tal-Singer R, and Klos A.. More Than Just Innate Immunity: Comparative Analysis of Chlamydophila Pneumoniae and


Chlamydia trachomatis Effects on Host-Cell Gene Regulation. Cell Microbiol. 2003 Nov; 5 (11).785-95 (50 citations)

PUBLISHED ABSTRACTS & PRESENTATIONS:

1. Su Y, Wang X, Jones E, Peters J, Mahdi OS, Zalduondo L, Stabenow J, Miyairi I, Lu L, Williams R, et al. Host genetics and upper genital tract disease in Chlamydia muridarum infected mice: forward genetic approach with translational implications. 13th International Symposium of Human Chlamydial Infections, Pacific Grove, CA, USA 2014 (oral presentation)

2. Peters J, and Byrne, GI. Cholesterol Growth Requirements for Chlamydia trachomatis Serovar D in HeLa Cells Are Dependent on Metabolic Activity by Host and Pathogen. 12th International Symposium on Human Chlamydial Infections, Salzburg, Austria June 2010 (oral presentation)

3. Peters J, Hsia R-c, and Bavoil PM. Late Type III Secreted Effector Protein CopN Targets Key Enzyme of Host Glycolytic Pathway. General Meeting of American Society for Microbiology, Boston, MA June 2008 (poster presentation)

4. Tall, EA, Peters, J., and Bavoil, PM. Identification of Type III Secretion Effectors in Chlamydia trachomatis Serovar D. General Meeting of American Society for Microbiology, Boston, MA June 2008 (poster presentation)

5. Peters J, Tatsuno I, Kaper J, and Bavoil PM. Establishment of Fluorescence-Based -Lactamase System for Identification of Chlamydial Type III Secretion Effector Proteins. General Meeting of American Society for Microbiology, Orlando, FL May 2006 (poster presentation)

6. Peters, J, Hess, S, Endlich, K, Thalmann, J, Bartling, G, and Klos, A. Hide-and-Seek: Host-Cell Responses in Models of Persistent Chlamydophila Pneumoniae Infection. 56. Conference of German Society for Hygiene and Microbiology, Münster, Germany September 2004 (poster presentation)

7. Peters, J, Hess, S, Endlich, K, Thalmann, J, Bartling, G, and Klos, A. Hide-and-Seek: Host-Cell Responses in Models of Persistent Chlamydophila Pneumoniae Infection. 5th Meeting of European Society for Chlamydia Research, Budapest, September 2004 (oral and poster presentation)

8. Endlich, K, Peters, J, Hess, S, Bartling, G, and Klos, A. Host-Cell Responses in Models of Persistent C. pneumoniae Infection; 55. Conference of German Society for Hygiene and Microbiology, Dresden, September 2003; published in International Journal of Medical Microbiology; 293, Suppl. No. 36: MP073 (poster presentation)

Jan Peters PhD May 2015 CV

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