Wieslaw Swietnicki, Ph.D.

E-mail: adpqem@r.postjobfree.com

Employment

1.2015-present

Institute of Immunology and Experimental Therapy of Polish Academy of Science ) ;

Position: Scientist/Project Leader

Address: ul. R. Weigla 12, 53-114 Wrocław, PL

2.2012-2015

Wroclaw Research Center EIT+ (currently PORT)

Position: Project Leader

Address: ul. Stablowicka 147, 54-066 Wroclaw, PL

3.2010-2011

Uniformed Health Services University

Position: Scientist

Address: 4301 Jones Bridge Rd., 20-814 Bethesda, MD, USA

4.2009-2010

Edgewood Chemical Biological Center

Position: Project Leader

Address: 5183 Blackhawk Rd., 21010-5424 Aberdeen Proving Ground, MD, USA

5.2000-2008

United States Army Medical Research Institute of Infectious Diseases

Position: Scientist/PI

Address: 1725 Porter Street, 21-702 Ft. Detrick, MD, USA

6.1996-2000

Case Western Reserve University

Position: Postdoctoral Fellow

Address: 10900 Euclid Ave, 44116-497 Cleveland, OH, USA

7.1989-1995

The University of Florida, School of Medicine, Department of Biochemistry and Molecular

Biology

Position: Graduate Assistant

Address: 1200 Newell Drive, 32610-0245 Gainesville, FL, USA

Academic and Research Career

A highly talented Protein Biochemist with vast experience and expertise in difficult protein expression, purification and biophysical characterization, including membrane proteins. Managerial experience (5 years) and proven ability to initiate projects, procure funding and guide multi-team efforts. Very energetic, open-minded and an excellent problem solver

driven by results. Coaching and training people (50+ people).

Early work included analysis of the structure-stability-pathogenicity relationship for human prion protein. The work led to the design of a stable protein incapable of conversion into a pathogenic form, theoretically offering a strategy to prevent prion diseases. Later work was conducted on the type III secretion system in Y. pestis and strategies to overcome antibiotic resistance. As a result, candidate therapeutics were identified and a live genetically-engineered safe vaccine was

constructed. The latest project included work on a virulence system from enteropathogenic E. coli which lead to safe and competitive inhibitors of a major virulence system in the pathogen.

Managerial experience

10 years in total, 2-3 people directly, 5+ indirectly

New lab design for biotechnology (USAMRIID, USA; ECBC, USA; EIT+, PL)

Selection of external service providers (molecular biology, synthetic organic chemistry, animal testing facilities)

Management of project costs and milestones, equipment procurement and maintenance

Identification of new strategies for project improvement

Preparation of progress reports

Management of IP issues, together with the IP Department, of new hits/leads

Development and coaching of the laboratory team

Citizenship: Poland, USA

Core Competencies:

Systems biology (analysis, de novo pathway design and assembly)

Cloning (GeneArt, Gateway, EK-LIC, RE), Site-directed Mutagenesis, Gene Knockout in Bacteria

manipulation (Vector NTI, standard gene tools)

Expression vector construction and Bacterial Recombinant Protein expression (E.coli, Yeast, Insect cells and Human Cells)

Protein Purification (AKTA FPLC/Explorer/Express, BioLogic DuoFlow), Column chromatography, protein purification automation

Antibody purification (monoclonal, polyclonal: protein A, immobilized antigen, regular chromatography)

Antibody-small molecule conjugation for imaging

Protein engineering to improve/remove receptor interactions

Design of stable formulations for novel vaccine candidates (hydrophobic/membrane proteins)

Cell cycle analysis (MUSE), mitochondrial potential interference (MUSE), apoptosis induction (MUSE: annexin V, caspase), MTT, Live/Dead cell count (MUSE)

Dichroism (Spectra Deconvolution, Tm, Cm)

Anisotropy for ligand binding, FRET, TR-FRET

Blotting and Co-immunoprecipitation, SDS-PAGE, ELISA

Labeling of Proteins, Chemical labeling of proteins (chemicals and fluorophores)

Surface Plasmon Resonance (Biacore)

Kinetics (UV/VIS, Fluorescence, FRET, TR-FRET) of proteins and small molecules

cGLP and GMP work (US)

Summary of Relevant Skills:

Synthetic biology (systems analysis, de novo pathway assembly, host’s metabolic system manipulation)

Ligand database preparation and HTS searching (Schroedinger/Sybyl-X + modeling suites)

Modeling (homology modeling, ligand binding, oligomer assembly)

Lead optimization (library design and screening, de novo ligand design)

QM (MM/QM, HF, LMP2, DFT, TDDFT) of small ligands, alone or in complexes

Professional Experience:

2015-present Institute of Imm. and Exp. Therapy, Wroclaw, Poland

Position Scientist

Responsibilities

Grant writing

Grant guidance

Antibacterial compounds development

2017-2022- design of a novel vaccine against a human pathogen- NCN-funded project

2012-2015 EIT+ Sp. Z o.o., Wroclaw, Poland

Position Project Leader

Responsibilities

Overall guidance of the projects

Coordination of a team effort to meet projects deadlines and objectives

Recruitment of new team members

Initiating new collaborations and project efforts

Presentation of work at internal and external meetings

Computer modeling, ligand database preparation, and HTS searching

Hit-to-lead and Lead optimization

Projects

1. Development of new inhibitors of enteropathogenic E. coli. The project has started in Jan 2012 and finished in 2015. The work led to the development of small molecule inhibitors of a specific bacterial enzyme essential for pathogen’s virulence. Best candidates were capable of blocking the infection in a cell culture model while showing minimal toxicity to mammalian cells. The lead candidate had Ki below 20 M. The strategy used a combination of HTS computational screening, biochemical assays for verification on a recombinant target, organic synthetic chemistry to increase potency and minimize toxicity, and cell culture studies to measure blockage of a biological function in a cell culture infection model.

Toxicity of compounds was measured in a cell culture model by looking at the interference with mitochondrial potential, apoptosis induction (caspase activation, Annexin V expression), cell cycle interference and viability by flow cytometry. The strategy can be used for many other human pathogens with known targets and available structural data. The approach is much more economical than the current commercial strategy based on the co-crystallization of target-inhibitor complexes and offers a time advantage when looking for potential new leads.

2. Development of efflux pump inhibitors for chemotherapy. The project started in March 2014 and led to the development of a hit candidate and identification of a different target. The target was verified on a recombinant protein.

The strategy used a known phenotypic hit and optimized it by exploring available chemical libraries. The best candidate had EC50 below 10 M and was capable of reversing drug resistance in chemotherapy-resistant cell line while showing minimal cellular toxicity.

The strategy used in the project is a part of the cheminformatics suite of tools employed in a commercial pharmaceutical industry. The approach is used when exploring known phenotypic hits and has an advantage over classical methods when looking for lead candidate identification.

Achievements

2 Patent applications submitted to others are in preparation. Details are confidential.

2010-2011 Uniformed Services University, Bethesda, MD

Position Staff

Project

Structure-function studies of selected human proteins.

Responsibilities

Protein expression, purification and characterization of multicomponent protein complexes

of proteins from tissues

Measurement of interactions (co-IP, co-expression, SEC, AUC)

Expression vector design

Achievements

Elucidation of mechanism for the connection between a human skeletal muscle disease

and a cellular degradation pathway

Design of a novel co-expression vector suitable for tandem affinity purification.

Measurement of physical interaction of proteins for the first time using the

designed vector.

2008- 2010 Research and Development Command-Edgewood Chemical Biological Center, Aberdeen Proving Ground, MD, USA

Position Sr. Research Scientist (Principal Investigator)

Project

Identification of small molecule inhibitors of bacterial type III secretion system ATPase.

Responsibilities

Guidance of the project and planning of team effort and individual members

Recruitment of potential collaborators

Securing funding for the future

Guidance of lab members in laboratory procedures if needed

Achievements

Design of engineered ATPase proteins suitable for inhibitor screening. The data were crucial for securing additional funding for equipment.

Identification of several inhibitors of ATPases with low micromolar IC50 in vitro assays and low M EC50 values in bacterial cell culture assays. Data were critical for securing the extension of the project funding into 2011-2012.

2003- 2008 U.S. Army Medical Research Institute of Infectious Diseases, Ft. Detrick, MD

Position Research Scientist

Project

Study of type III secretion system protein-protein interactions.

Responsibilities

Guidance of the project and planning of team effort and individual members potential collaborators

Funding for the future

Guidance of lab members in laboratory procedures if needed

Achievements

Funding for the project and organizing the team

of proof-of-principle of the proposed strategy in an animal model

Project

Investigation of novel therapeutic targets for the blockage of the type III secretion system from Y. pestis.

Responsibilities

Guidance of technicians on the project

Help with protein-protein interaction measurements

Achievements

Identification of a novel approach for delineating protein-protein interactions in an unknown system

Identification of the mechanism of early steps in protein secretion by Y. pestis.

2000-2003 U.S. Army Medical Research Institute of Infectious Diseases,

Ft. Detrick, MD, USA

Position Senior Research Fellow

Project

Elucidation of the mechanism of human immune system inactivation by Spe-C superantigen from S. pyogenes

Responsibilities

Conceptual guidance of the project

Protein production

Measurement of interactions of isolated proteins with recombinant other proteins or cell surface antigens

Docking of ligands and proteins to other proteins

Achievements

Experimental confirmation of a novel mechanism for coupling between MHC class II and T-cell receptor molecules of a recombinant vaccine for S. pyogenes

1996- 2000 Case Western Reserve University, Cleveland, OH

Position Postdoctoral Fellow

Project

Identification of a link between structure and pathogenicity of human prion protein

Responsibilities

Protein expression, purification, and characterization

Achievements

Structural and biophysical verification of aggregation as the cause of formation of an abnormal form of human prion protein

In vivo reconstruction of conditions leading to the formation of aggregated prion protein variants

.

1988- 1989 Mann Testing Laboratories, Mississauga, ON, Canada

Position GC/MS Operator

Responsibilities

GC/MS analysis of molecules

Achievements

Writing of a program for fully automatic sample analysis and equipment validation

1984- 1985 Wroclaw Inorganic Works, Wroclaw, Poland

Position Head of QC laboratory

Responsibilities

Laboratory guidance

Design and implementation of SOPs and regulatory documents for the analytical chemistry laboratory

Achievements

Laboratory performance improvement

Education and Professional Training:

1989-1995 - Ph.D. in Biochemistry and Molecular Biology, University of Gainesville, FL. Thesis: Enzyme mechanisms

and kinetic analysis of a Hepatitis A Virus 3 C protease, a viral protein processing enzyme.

1976-1981 - M.Sc. in Chemistry with conc. in Physical Chemistry, M. Sc., University of Wroclaw, Poland

Honors and Distinctions:

2006 Invited Speaker for Protein Engineering Group, Cambridge, MA

2001-2003 Awarded the National Science Foundation Post-doctoral Fellowship ($50,000/year for 3 years)

2008-2010 Awarded the Defense Threat Reduction Agency Grant ($1 million/3 years) Role: PI

2010 In-house ILIR project ($150K/year) Role: co-PI

2011-2012 Awarded the Defense Threat Reduction Agency Grant ($1 million/2 years) Role: PI

2012-2014 Awarded BioMed grant (1.5 mln PLN/3 years)

Role: PI

2013-2014 Awarded BioMed grant (550K PLN/1 year)

Role: PI

2014 Invited Speaker for In Silico Drug Discovery Conference, Durham, NC

2017-2022 Awarded National Science Foundation grant (1,300K PLN/3 years)

Role: PI

2018 Invited Speaker for GE Healthcare meeting, Serock, PL

2018 Speaker, Institute of Immunology and Experimental Therapy PAS, Wroclaw, PL

2018 Organizer and Speaker, International Conference on Computer-Aided Drug Design, Institute of Immunology and Experimental Therapy PAS, Wroclaw, PL

2019 Organizer and Speaker, 2nd International Conference on Computer-Aided Drug Design, Institute of Immunology and Experimental Therapy PAS, Wroclaw, PL

Publications:

Total number of citations: 2156

Hirsch Index: 19

Publications

[1-32]

1. Swietnicki, W.; Goldeman, W.; Psurski, M.; Nasulewicz-Goldeman, A.; Boguszewska-Czubara, A.; Drab, M.; Sycz, J.; Goszczyński, T.M. Metallacarborane Derivatives Effective against Pseudomonas aeruginosa and Yersinia enterocolitica. Int J Mol Sci 2021, 22, doi:10.3390/ijms22136762.

2. Swietnicki, W.; Caspi, R. Prediction of Selected Biosynthetic Pathways for the Lipopolysaccharide Components in Porphyromonas gingivalis. Pathogens 2021, 10, 374.

3. Swietnicki, W. Secretory System Components as Potential Prophylactic Targets for Bacterial Pathogens. Biomolecules 2021, 11, doi:10.3390/biom11060892.

4. Chatterjee, P.; Sass, G.; Swietnicki, W.; Stevens, D.A. Review of Potential Pseudomonas Weaponry, Relevant to the Pseudomonas-Aspergillus Interplay, for the Mycology Community. J Fungi (Basel) 2020, 6, doi:10.3390/jof6020081.

5. Swietnicki, W.; Czarny, A.; Antkowiak, L.; Zaczynska, E.; Kolodziejczak, M.; Sycz, J.; Stachowicz, L.; Alicka, M.; Marycz, K. Identification of a potent inhibitor of type II secretion system from Pseudomonas aeruginosa. Biochem Biophys Res Commun 2019, 513, 688-693, doi:10.1016/j.bbrc.2019.04.055.

6. Swietnicki, W.; Brzozowska, E. In silico analysis of bacteriophage tail tubular proteins suggests a putative sugar binding site and a catalytic mechanism. J Mol Graph Model 2019, 92, 8-16, doi:10.1016/j.jmgm.2019.07.002.

7. Swietnicki, W.; Czarny, A.; Urbanska, N.; Drab, M. Identification of small molecule compounds active against Staphylococcus aureus and Proteus mirabilis. Biochem Biophys Res Commun 2018, 506, 1047-1051, doi:10.1016/j.bbrc.2018.10.189.

8. Bzdzion, L.; Krezel, H.; Wrzeszcz, K.; Grzegorek, I.; Nowinska, K.; Chodaczek, G.; Swietnicki, W. Design of small molecule inhibitors of type III secretion system ATPase EscN from enteropathogenic Escherichia coli. Acta Biochim Pol 2017, 64, 49-63, doi:10.18388/abp.2016_1265.

9. Milczarek, M.; Filip-Psurska, B.; Swietnicki, W.; Kutner, A.; Wietrzyk, J. Vitamin D analogs combined with 5-fluorouracil in human HT-29 colon cancer treatment. Oncol Rep 2014, 32, 491-504, doi:10.3892/or.2014.3247.

10. Kong, Q.; Mills, J.L.; Kundu, B.; Li, X.; Qing, L.; Surewicz, K.; Cali, I.; Huang, S.; Zheng, M.; Swietnicki, W., et al. Thermodynamic stabilization of the folded domain of prion protein inhibits prion infection in vivo. Cell Rep 2013, 4, 248-254, doi:10.1016/j.celrep.2013.06.030.

11. Sambuughin, N.; Swietnicki, W.; Techtmann, S.; Matrosova, V.; Wallace, T.; Goldfarb, L.; Maynard, E. KBTBD13 interacts with Cullin 3 to form a functional ubiquitin ligase. Biochem Biophys Res Commun 2012, 421, 743-749, doi:10.1016/j.bbrc.2012.04.074.

12. Bozue, J.; Cote, C.K.; Webster, W.; Bassett, A.; Tobery, S.; Little, S.; Swietnicki, W. A Yersinia pestis YscN ATPase mutant functions as a live attenuated vaccine against bubonic plague in mice. FEMS Microbiol Lett 2012, 332, 113-121, doi:10.1111/j.1574-6968.2012.02583.x.

13. Swietnicki, W.; Carmany, D.; Retford, M.; Guelta, M.; Dorsey, R.; Bozue, J.; Lee, M.S.; Olson, M.A. Identification of small-molecule inhibitors of Yersinia pestis Type III secretion system YscN ATPase. PLoS One 2011, 6, e19716, doi:10.1371/journal.pone.0019716.

14. Raab, R.; Swietnicki, W. Yersinia pestis YopD 150-287 fragment is partially unfolded in the native state. Protein Expr Purif 2008, 58, 53-60, doi:10.1016/j.pep.2007.11.001.

15. Swietnicki, W. Folding aggregated proteins into functionally active forms. Curr Opin Biotechnol 2006, 17, 367-372, doi:10.1016/j.copbio.2006.05.011.

16. Hale, M.L.; Swietnicki, W. Model systems to study a superantigen-induced disease: Toxic shock syndrome. Drug Discovery Today: Disease Models 2006, 3, 121-126, doi:https://doi.org/10.1016/j.ddmod.2006.03.015.

17. Swietnicki, W.; Powell, B.S.; Goodin, J. Yersinia pestis Yop secretion protein F: purification, characterization, and protective efficacy against bubonic plague. Protein Expr Purif 2005, 42, 166-172, doi:10.1016/j.pep.2005.02.016.

18. Swietnicki, W.; O'Brien, S.; Holman, K.; Cherry, S.; Brueggemann, E.; Tropea, J.E.; Hines, H.B.; Waugh, D.S.; Ulrich, R.G. Novel protein-protein interactions of the Yersinia pestis type III secretion system elucidated with a matrix analysis by surface plasmon resonance and mass spectrometry. J Biol Chem 2004, 279, 386**-*****, doi:10.1074/jbc.M405217200.

19. Swietnicki, W.; Barnie, A.M.; Dyas, B.K.; Ulrich, R.G. Zinc binding and dimerization of Streptococcus pyogenes pyrogenic exotoxin C are not essential for T-cell stimulation. J Biol Chem 2003, 278, 9885-9895, doi:10.1074/jbc.M206957200.

20. Derrington, E.; Gabus, C.; Leblanc, P.; Chnaidermann, J.; Grave, L.; Dormont, D.; Swietnicki, W.; Morillas, M.; Marck, D.; Nandi, P., et al. PrPC has nucleic acid chaperoning properties similar to the nucleocapsid protein of HIV-1. C R Biol 2002, 325, 17-23, doi:10.1016/s1631-0691(02)01388-4.

21. Knaus, K.J.; Morillas, M.; Swietnicki, W.; Malone, M.; Surewicz, W.K.; Yee, V.C. Crystal structure of the human prion protein reveals a mechanism for oligomerization. Nat Struct Biol 2001, 8, 770-774, doi:10.1038/nsb0901-770.

22. Gabus, C.; Derrington, E.; Leblanc, P.; Chnaiderman, J.; Dormont, D.; Swietnicki, W.; Morillas, M.; Surewicz, W.K.; Marc, D.; Nandi, P., et al. The prion protein has RNA binding and chaperoning properties characteristic of nucleocapsid protein NCP7 of HIV-1. J Biol Chem 2001, 276, 193**-*****, doi:10.1074/jbc.M009754200.

23. Gabus, C.; Auxilien, S.; Pechoux, C.; Dormont, D.; Swietnicki, W.; Morillas, M.; Surewicz, W.; Nandi, P.; Darlix, J.L. The prion protein has DNA strand transfer properties similar to retroviral nucleocapsid protein. J Mol Biol 2001, 307, 1011-1021, doi:10.1006/jmbi.2001.4544.

24. Zhang, Y.; Swietnicki, W.; Zagorski, M.G.; Surewicz, W.K.; Sonnichsen, F.D. Solution structure of the E200K variant of human prion protein. Implications for the mechanism of pathogenesis in familial prion diseases. J Biol Chem 2000, 275, 336**-*****, doi:10.1074/jbc.C000483200.

25. Swietnicki, W.; Morillas, M.; Chen, S.G.; Gambetti, P.; Surewicz, W.K. Aggregation and fibrillization of the recombinant human prion protein huPrP90-231. Biochemistry 2000, 39, 424-431, doi:10.1021/bi991967m.

26. Li, R.; Liu, T.; Wong, B.S.; Pan, T.; Morillas, M.; Swietnicki, W.; O'Rourke, K.; Gambetti, P.; Surewicz, W.K.; Sy, M.S. Identification of an epitope in the C terminus of normal prion protein whose expression is modulated by binding events in the N terminus. J Mol Biol 2000, 301, 567-573, doi:10.1006/jmbi.2000.3986.

27. Morillas, M.; Swietnicki, W.; Gambetti, P.; Surewicz, W.K. Membrane environment alters the conformational structure of the recombinant human prion protein. J Biol Chem 1999, 274, 368**-*****, doi:10.1074/jbc.274.52.36859.

28. Swietnicki, W.; Petersen, R.B.; Gambetti, P.; Surewicz, W.K. Familial mutations and the thermodynamic stability of the recombinant human prion protein. J Biol Chem 1998, 273, 310**-*****, doi:10.1074/jbc.273.47.31048.

29. Swietnicki, W.; Petersen, R.; Gambetti, P.; Surewicz, W.K. pH-dependent stability and conformation of the recombinant human prion protein PrP(90-231). J Biol Chem 1997, 272, 275**-*****, doi:10.1074/jbc.272.44.27517.

30. Johansson, P.J.; Malone, C.; Swietnicki, W.; Dunn, B.M.; Williams, R.C., Jr. Fv structure of monoclonal antibody II-481 against herpes simplex virus Fc gamma-binding glycoprotein gE contains immunodominant complementarity determining region epitopes that react with human immunoglobulin M rheumatoid factors. J Exp Med 1994, 180, 1873-1888, doi:10.1084/jem.180.5.1873.

31. Dunn, B.M.; Scarborough, P.E.; Davenport, R.; Swietnicki, W. Analysis of proteinase specificity by studies of peptide substrates. The use of UV and fluorescence spectroscopy to quantitate rates of enzymatic cleavage. Methods Mol Biol 1994, 36, 225-243, doi:10.1385/0-89603-274-4:225.

32. Jewell, D.A.; Swietnicki, W.; Dunn, B.M.; Malcolm, B.A. Hepatitis A virus 3C proteinase substrate specificity. Biochemistry 1992, 31, 7862-7869, doi:10.1021/bi00149a017.

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