2019 RADD Posters and Core Facilities

"Core Facilities" exhibitors (i.e., facilities available for users external to the institution) are indicated by "►" preceding the listing. Brief listing of all exhibitors is below at top of page. Detailed abstracts appear in longer list below the brief list.

 

1. Subhadwip Basu, Rutgers University

Design, Synthesis, Evaluation and X-ray Structure of Small Molecule Inhibitors of the Programmed Cell Death-1 (PD-1)/Programmed Death-Ligand 1 (PD-L1) Protein-Protein Interaction. Presenter contact: subhadwip.basu@rutgers.edu

2. Rajesh Singh, Abzyme Therapeutics

Single domain antibodies as alternative antibodies in the next generation immunotherapy. Presenter contact: Singh@abzymetx.com

►3. Shoreh Miller, Rutgers University

In Vivo Research Services (IVRS), Rutgers Presenter contact: shoreh.miller@rutgers.edu

4. Qian Xie, Princeton University

RaPID discovery of novel macrocyclic peptide inhibitors of S. aureus quorum sensing-driven virulence. Presenter contact: qx10@princeton.edu

5. Wen Kang Chou, Princeton University

Small Molecule Screening for Inhibitors of Escherichia coli Nitric Oxide Defense. Presenter contact: wkchou@princeton.edu      

6. Amelia Dunn, Rockefeller University

Correlation between Beta-Arrestin-2 signaling and rotarod sedation at the kappa opioid receptor. Presenter contact: adunn@rockefeller.edu

7. Youyi Peng, Rutgers University

Computer-Aided Discovery of Sigma 1 Receptor Antagonists as Novel Treatments for Neuropathic Pain. Presenter contact: pengyo@cinj.rutgers.edu

8. Jacob A. Kautzky, Princeton University

Streamlining Drug Discovery through Development of a Novel C-C Bond Formation. Presenter contact: jkautzky@princeton.edu

9. Allen B. Reitz, Fox Chase Chemical Diversity Center, Inc.

Characterization of small-molecule ligands that bind to monomeric TDP-43 directly and prolong life and ameliorate locomotor dysfunction in human WT and mutant TDP-43 Drosophil. Presenter contact: areitz@fc-cdci.com

10. Troy E. Messick, The Wistar Institute

Development of Small Molecule Inhibitors of EBNA1 for the Treatment of Nasopharyngeal Carcinoma. Presenter contact: tmessick@wistar.org

11. Goodwell Nzou, Wake Forest University, currently University of Pennsylvania

Three Dimensional Model of the Blood Brain Barrier for Drug Discovery and Disease Modeling. Presenter contact: nzoug@pennmedicine.upenn.edu     

12. Joseph P. Sheehan, Princeton University

A dual-mechanism antibiotic targets Gram-negative bacteria and avoids drug resistance. Presenter contact: jpsheehan@princeton.edu

13. Doreen Badheka, Rutgers University

Teaching the pharmaceutical research, approval and regulatory processes related to drug discovery. Presenter contact: doreen.badheka@rutgers.edu

14. Kyle S. Saitta, Rutgers University

The Selective Group I Metabotropic Glutamate Receptor Agonist 2-Chloro-5-hydroxyphenylglycine (CHPG) Enhances BDNF and Reverses Demyelination. Presenter contact: kyle.saitta@rutgers.edu

15. Maxim Chudaev, Rutgers University

Engineered EF-Tu and tRNA in FRET-based screening assay for identification of inhibitors of protein synthesis in bacteria. Presenter contact: chudaymv@rutgers.edu

16. Mark C. Siracusa, Rutgers University

Targeting carbonic anhydrase enzymes to prevent mast cell development and mastocytosis. Presenter contact: mark.siracusa@rutgers.edu

17. Salvatore Coniglio, Kean University

Using CCR1 Antagonists to Target Tumor Associated Macrophages. Presenter contact: coniglsa@kean.edu

►18. Research Pathology Services

 Michael Goedken, Rutgers University. Presenter contact: michael.goedken@rutgers.edu

19. Longqin Hu, Rutgers University

L-Cystine Diamides as a Novel Therapy for Cystinuria. Presenter contact: longhu@pharmacy.rutgers.edu

►20. Molecular Design and Synthesis Group Collaborations, Rutgers University Biomedical Research Innovation Cores (RUBRIC).

Jacques Y. Roberge, Rutgers University. Presenter contact: jr1257@ored.rutgers.edu

21. Ila Nimgaonkar, Princeton University

Isocotoin potently inhibits hepatitis E virus replication through interference with heat shock-protein 90. Presenter contact: inimgaon@princeton.edu

22. Chang Tian, Princeton University

Translation of Nanoparticle Therapeutics from Laboratory Discovery to Clinical Application: Scaled-up production and processing. Presenter contact: ctian@princeton.edu

23. Chester Markwalter, Princeton University

Protein and Peptide Formulations in Highly-Loaded Nanocarriers. Presenter contact: cem3@princeton.edu

24. Makayla Pardo, Drew University

Reactivation of mutant p53 by small molecules for potential cancer therapeutics. Presenter contact: mpardo@drew.edu

25. Julie S. Valastyan and Isabelle R. Taylor, Princeton University

Discovery of PqsE Thioesterase Enzyme Inhibitors for the Bacterial Pathogen Pseudomonas aeruginosa using DNA-Encoded Small Molecule Library Screening. Presenter contacts: juliev@princeton.edu, itaylor@princeton.edu

►26. Cell and Cell Products Fermentation Facility

Arvin Lagda, Rutgers University. Presenter contact: arvin.lagda@waksman.rutgers.edu

►27. Selected Applications by NMR

István Pelczer, Princeton University. Presenter contact: ipelczer@princeton.edu

►28. The Prism Imaging and Analysis Center − An engine for innovation and discovery 

 Nan Yao, Princeton University. Presenter contact: nyao@princeton.edu

►29. The Biological Mass Spectrometry Facility at CABM, Rutgers 

Haiyan Zheng, Rutgers University. Presenter contact: Haiyanz@cabm.rutgers.edu 

►30. Proteomics and Mass Spectrometry Core Facility

Saw Kyin, Princeton University. Presenter contact: skyin@princeton.edu 

►31. Princeton University Flow Cytometry Resource Facility.

Christina J. DeCoste, Princeton University. Presenter contact: cdecoste@princeton.edu 

32. Sandra J. Aedo, Princeton University

Nitrofurantoin tolerance of non-growing Escherichia coli and their re-sensitization.  Presenter contact: saedo@princeton.edu 

33. Shiva V. Patnala, William Paterson University

Application of the Clustering of Ligand Diffusion Coefficient Pairs (CoLD-CoP) Method to Detecting Cooperatively Binding Drug Fragments. Presenter contact: PatnalaV@student.wpunj.edu

34. Allison Murawski and Mark Brynildsen, Princeton University

Ploidy Impacts Persistence to Fluoroquinolones. Presenter contact: allisonm@princeton.edu 

 

 

Abstracts

Poster 1

Design, Synthesis, Evaluation and X-ray Structure of Small Molecule Inhibitors of the Programmed Cell Death-1 (PD-1)/Programmed Death-Ligand 1 (PD-L1) Protein-Protein Interaction

Subhadwip Basu1, Jeffrey Yang1, Bin Xu1, Vladyslav Kholodovych1,2, Katarzyna Magiera-Mularz3, Bogdan Musielak3, Tad A. Holak3, Longqin Hu1,4

 

1 Department of Medicinal Chemistry, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, Piscataway, NJ 08854

2 High Performance and Research Computing, Office of Advanced Research Computing, Rutgers, The State University of New Jersey, Piscataway, NJ 08854

3 Department of Organic Chemistry, Faculty of Chemistry, Jagiellonian University, 30-060 Krakow, Poland

4 The Cancer Institute of New Jersey, New Brunswick, NJ 08901

 

The PD-1/PD-L1 immune checkpoint pathway is a clinically validated drug target engaged in the suppression of tumor immunity. Despite the clinical success of antibodies in patients with advanced cancers, there are currently no FDA-approved small molecule immunomodulators for this immune checkpoint pathway. Small molecules offer an appropriate pharmacokinetic profile for oral administration, increased tumor penetration, and shorter half-life for management of intractable immune-related side effects. Here, we present our design, synthesis, and a preliminary structure-activity relationship study of a series of C2-symmetric inhibitors. Based on structural studies, the C2-symmetric inhibitor LH1307 (2b) directly bound to PD-L1 and induced the formation of a near symmetrically arranged PD-L1 homodimer. LH1307 demonstrated an IC50 of 3 nM in a homogeneous time resolved fluorescence assay. In a cell-based PD-1/PD-L1 blockade assay, LH1306 (2a) and LH1307 were found to be about 8.2- and 2.8-fold more potent in inhibiting the PPI as compared to LH1301 (1a) and LH1305 (1b), respectively. The new C2-symmetric inhibitors could serve as new leads for further optimization into potential immunomodulators for the treatment of cancer.

Presenter contact: subhadwip.basu@rutgers.edu

 


Poster 2

Single domain antibodies as alternative antibodies in the next generation immunotherapy

 

F. Moussa1, C. Campbell1, A. Pandya1, B. Lee1, H. Tran1, R. Singh1

 

1 Abzyme Therapeutics, 321 Jones Boulevard, Royersford, PA 19468

 

Monoclonal antibodies are used for the treatment of a wide range of diseases from cancer to infectious diseases. The discovery of antibodies lacking light chains in camelids has spurred the development of single-domain VHH antibodies for therapeutic and diagnostic applications. The advantages of VHHs include small-size, thermally stable, and capable of recognizing antigenic sites that are normally not accessible to conventional antibodies. The ability of VHH to recognize these recessed antigenic epitopes has been attributed to their smaller size and the ability of their extended CDR3 loop to penetrate into cavities on the antigens. The approval of the first therapeutic VHH directed against the von Willebrand factor for the treatment of acquired thrombotic thrombocytopenic purpura (Caplacizumab by Ablynx), is expected to bolster the rise of these innovative molecules. Using Abzyme’s proprietary SuperLlama/SuperYeast high-throughput approach for VHH antibody discovery, we have successfully isolated numerous VHH antibodies against a few dozen [HH1] therapeutic and diagnostic biomarkers. Here, we present data related to the successful development of an ANTAGONIST and AGONIST VHH antibodies against two therapeutically important targets, respectively. Since these two targets are therapeutically synergistic, we constructed a series of bi-specific bivalents (2+2) and our pre-clinical data showed that they are well-expressed, highly stable, and functionally better than the best-in-class mono-specific bivalents. We are currently looking for pharmaceutical collaborators to further develop and advance these antibodies into clinical trials.

 

Presenter contact: Singh@abzymetx.com

 

Poster 3

In Vivo Research Services (IVRS), Rutgers
Shoreh Miller.

 

The mission of the IVRS is to provide centralized high quality, cost-effective research services to both academic and industry clients.  Rutgers has a wealth of experience, capabilities and models that are routinely used to answer critical in vivo research questions. We provide a comprehensive program of animal care including consultation, experimental design, protocol preparation, a duly constituted animal care and use committee, occupational health and laboratory safety, BSL3 in vivo biohazard research, and full-time veterinary care.  Rutgers offers competitive fee-for-service pricing, confidentiality, accuracy, and rapid service. Our team is highly experience and dedicated to humane animal research.

 

Presenter contact: shoreh.miller@rutgers.edu

 


Poster 4

 

RaPID discovery of novel macrocyclic peptide inhibitors of S. aureus quorum sensing-driven virulence

 

Qian Xie1, Aishan Zhao1, Mareike Wiedmann2, Richard Novick3, Hiroaki Suga2, Tom Muir1

 

1 Department of Chemistry, Princeton University, Princeton, NJ 08544

2 Department of Chemistry, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, Japan

3 Skirball Institute, Department of Microbiology, NYU Medical Center, New York, NY 10016

 

Staphylococcus aureus infections present as a serious threat to public health, especially with the rise of antibiotic resistance. In looking for an alternative target for therapeutic intervention of Staph infections, we focused our attention on the agr quorum sensing pathway, as agr activation in S. aureus leads to the production of myriad virulence factors that cause infection symptoms. Specifically, we demonstrated here the discovery of novel macrocyclic peptide inhibitors targeting the receptor histidine kinase AgrC involved in agr signaling, using a combination of the lipid nanodisc reconstitution technology and an mRNA display-based selection system (RaPID system). The peptide hits emerged from the RaPID system showed potent competitive inhibition of AgrC in vitro and effective suppression of Staph virulence in vivo. We envisioned that the same approach could also be extended to other pharmacologically important membrane proteins, thereby creating ample opportunities for drug discovery.

 

Presenter contact: qx10@princeton.edu

 

Poster 5

Small Molecule Screening for Inhibitors of Escherichia coli Nitric Oxide Defense

Wen Kang Chou and Mark P. Brynildsen


Princeton University, 215 Hoyt Laboratory, William Street, Princeton NJ 08540

 

The development of antibiotic resistance poses a serious threat to global health and modern medicine. Antibiotics are used not only to treat infectious diseases but also as a precaution to prevent complications after invasive medical procedures. Over time, bacteria have developed and disseminated defenses against these live-saving drugs, leading to increasing failures of antibiotic treatments, which include last resort treatments. To try to assuage this advancing health crisis, we sought to develop novel anti-infectives by targeting pathogens’ virulence, and specifically their defenses against nitrosative stress. Nitric oxide (NO) is synthesized by phagocytes, which constitute the body’s first line of defense, to eliminate invading bacteria. However, various pathogens have developed defenses against nitric oxide, allowing them to withstand immune attack, proliferate in the host and ultimately cause an illness. We aimed to identify inhibitors of bacterial NO defenses by screening for molecules that impede bacterial growth in the presence of NO, but not its absence. Of more than 8,000 compounds screened, we found 4 hits that hold promising properties. One of them, 2-mercaptobenzothiazole (2-MBT), is especially interesting because it can nearly completely thwart the aerobic NO detoxification in Escherichia coli, while not significantly undermining growth under normal growth conditions. Preliminary results suggest that 2-MBT is a direct inhibitor of Hmp, the enzyme responsible for detoxifying NO under oxygenated conditions in E. coli. Current effort is focusing on elucidating the mechanism of action of 2-MBT, and studying the potency of its analogs to identify chemical components that are essential for its inhibitory effects.

 

Presenter contact: wkchou@princeton.edu

 

Poster 6

 

Correlation between Beta-Arrestin-2 signaling and rotarod sedation at the kappa opioid receptor

 

Amelia Dunn, Brian Reed, Jose Erazo, Ariel Ben-Ezra, Mary Jeanne Kreek

 

The Laboratory of the Biology of Addictive Diseases, Rockefeller University, 1230 York Ave, New York, NY 10065

 

The kappa opioid receptor (KOR) is an important drug target for diseases of mood and reward, such as addiction and depression. KOR agonists have been shown to block drug-seeking in animal models, however they also cause significant side effects such as sedation and aversion. In order to preserve the therapeutic effects of KOR agonists while avoiding negative side effects, many groups are investigating biased agonists that selectively activate only certain pathways downstream of the receptor. We compared the signaling properties of 21 KOR agonists, including the endogenous peptide Dynorphin A (1-17), morphinan compounds approved for human use, and 2 novel compounds synthesized for our research. We found that most compounds tested were biased for G-protein signaling over Beta-Arrestin-2 recruitment compared to a reference KOR agonist. A subset of these compounds was tested for KOR-mediated sedation using the rotarod assay in mice. We found a significant correlation between maximum effect on the rotarod and maximum efficacy in the Beta-Arrestin-2 recruitment assay, but not the G-protein signaling assay. This suggests that Beta-Arrestin-2 signaling in this in vitro system can model in vivo sedation behavior, and that a G-protein biased KOR agonist may not cause the sedative side effects associated with full, unbiased KOR activation.

 

Presenter contact: adunn@rockefeller.edu

 

Poster 7

 

Computer-Aided Discovery of Sigma 1 Receptor Antagonists as Novel Treatments for Neuropathic Pain

 

Youyi Peng1 and William Welsh2

 

1 Biomedical Informatics Shared Resources, Rutgers Cancer Institute of New Jersey, 195 Little Albany Street, New Brunswick, NJ 08903

2 Department of Pharmacology, Rutgers Robert Wood Johnson Medical School, 661 Hoes Lane, Piscataway, NJ 08854

 

The sigma 1 receptor (S1R) is a ligand-gated molecular chaperone protein within the endoplasmic reticulum and plasma membranes, and has been implicated in various pathological disorders including neuropathic pain. S1R antagonists have been shown to attenuate neuropathic pain of different etiology in rodent and human, including chemotherapy-induced and streptozotocin (STZ)-induced diabetic neuropathic pain. Guided by quantitative structure-activity relationships (QSAR) and molecular docking, we have discovered a novel family of S1R antagonists that exhibit potent binding affinity and high selectivity for S1R, favorable in vitro ADME/Tox properties and in vivo pharmacokinetics with high blood-brain permeability (B/P ratio = 14). Our lead compound PW507 has demonstrated exceptional antinociceptive activity for chemotherapy (taxol)-induced pain and STZ-induced diabetic pain without causing motor incoordination in rats. Further in vivo efficacy studies to assess the potential of modifying disease progression in diabetic db/db mice is currently underway.

 

Presenter contact: pengyo@cinj.rutgers.edu



Poster 8

 

Streamlining Drug Discovery through Development of a Novel C-C Bond Formation

 

Jacob A. Kautzky, Tao Wang, Ryan W. Evans, David W. C. MacMillan

 

Merck Center for Catalysis at Princeton University, Princeton, New Jersey 08544, United States

 

The merger of transition metal catalysis and photoredox catalysis has enabled the discovery of a range of synthetically frustrating and hitherto unknown transformations. Use of carboxylic acids in particular have revolutionized the pharmaceutical industry due to their being relatively abundant native functionality that can be diversified to a variety of moieties. Building upon previous work, a novel decarboxylative C—C bond formation is accomplished via metallaphotoredox catalysis. This transformation is utilized for the late stage functionalization of over ten natural products and pharmaceutical drugs.

 

Presenter contact: jkautzky@princeton.edu

 

Poster 9

 

Characterization of small-molecule ligands that bind to monomeric TDP-43 directly and prolong life and ameliorate locomotor dysfunction in human WT and mutant TDP-43 Drosophila

 

Marianne Carlsen1, Alyssa Coyne2, Randall Binder1, Xiao Jin2, Richard W. Scott1, Katie B. Freeman1, Amy Banks1, Samantha DeSando1, Ben Zaepfel2, Daniela C. Zarnescu2 and Allen B. Reitz1

 

1 Fox Chase Chemical Diversity Center, Inc., Pennsylvania Biotechnology Center, 3805 Old Easton Rd., Doylestown, PA 18902

 

2  Departments of Molecular and Cellular Biology, and Neurology, University of Arizona, Life Sciences South Building, 1007 E Lowell Street, Tucson, AZ 85721

 

The heterogeneous ribonucleoprotein TAR DNA binding protein 43 (TDP-43) is associated with neurodegeneration in diseases such as amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD-TDP-43). Here we examine the effects of small-molecule ligands with affinity for monomeric TDP-43 identified by screening of a diverse compound library for binding to TDP-43 using an alpha-screen assay followed by iterative SAR development preparing >400 new chemical entities increasing potency to 20-30 nM IC50 values. Early lead FC-776 prolongs life and ameliorates locomotor dysfunction in Drosophila expressing human WT or mutant TDP-43. 

 

Presenter contact: areitz@fc-cdci.com

 

Poster 10

 

Development of Small Molecule Inhibitors of EBNA1 for the Treatment of Nasopharyngeal Carcinoma

 

Troy E. Messick1, Samantha Soldan1, Garry R Smith2, Kimberly Malecka1, Lois Tolvinski1, Julianna S. Deakyne1, A.J. Pieter van den Heuvel3, Bai-Wei Gu3, Joel Cassel1,3, Mark McDonnell2, Yang Zhang2, Marianne Carlsen2, Shuai Chen2, Allen B. Reitz2, Paul M. Lieberman1

 

1 The Wistar Institute, 3601 Spruce Street, Philadelphia, PA 19104

2 Fox Chase Chemical Diversity Center, Inc., Pennsylvania Biotechnology Center

3805 Old Easton Rd., Doylestown, PA  18902

3 Vironika, LLC, 100 E. Lancaster Ave, LIMR Rm 133, Wynnewood, PA 19096

 

Epstein-Barr virus Nuclear Antigen 1 (EBNA1) is a multifunctional dimeric protein critical for viral replication, genome maintenance and viral gene expression.  We used a fragment-based approach to develop a small molecule lead series that selectively inhibits the DNA-binding activity of EBNA1.  Using computational chemistry, NMR screening and Surface Plasmon Resonance screening, we selected fragments for further structural elucidation.  We determined the structures of 22 different fragments at 4 different sites on EBNA1.  The lead series was discovered when we merged two fragments together to achieve potency greater than the individual fragments.   We characterized the individual members of the lead series in several cell-based assays, including ChIP assays, and we demonstrated that these inhibitors provide protection in xenograft models of EBV-driven tumor growth of NPC cell lines. Furthermore, EBER-ISH experiments confirm in vivo target engagement by the elimination of EBV in treated tumor tissue. The lead series meet or exceed industry-accepted criteria for drug suitability, safety and toxicology including physicochemical properties, metabolic stability, selectivity in broad-based screens and bioavailability, and the absence of in vivo liabilities, including 28-day toxicity and genotoxicity studies.  We submitted an Investigational New Drug (IND) application and received a “Study May Proceed” letter from the U.S. FDA, ClinicalTrials.gov Registry: NCT03682055.  The Phase I clinical trial is ongoing. These data establish proof-of-concept for targeting EBNA1—a protein previously thought to be undruggable.

 

Presenter contact: tmessick@wistar.org


Poster 11

 

Three Dimensional Model of the Blood Brain Barrier for Drug Discovery and Disease Modeling

 

Goodwell Nzou, N. VanOstrand, R.T. Wicks, E.E. Wicks, S. A. Seale, C. H. Sane, A. Chen. S.V. Murphy, J.D. Jackson, A.J. Atala

 

Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, 391 Technology Way, Winston-Salem, NC  27101

 

The blood brain barrier (BBB) is a dynamic component of the brain that prevents entry of foreign substances into the brain parenchyma. Consisting primarily of 3 cell types, endothelial cells, pericytes, and astrocytes, the BBB maintains brain homeostasis and low permeability through the expression tight junction (TJ) and adherent junction (AJ) proteins by endothelial cells. The integral selectivity characteristic of the BBB limits therapeutic options for many neurologic diseases and disorders. Currently, very little is known about the mechanisms that govern the dynamic nature of BBB. Furthermore, most in vitro models only utilize endothelial cells, pericytes and astrocytes. These models neglect the role of other cell types in the brain cortex such as the neurons, microglia and oligodendrocytes. Thus, we created a 3D organoid model the BBB consisting of six major cell types that closely recapitulate normal human brain tissue. The organoids were assessed for BBB properties. Our data demonstrates the formation of both tight junctions and adherens junctions in organoids. In addition, our data on BBB functionality assessment using MPTP, MPP+ and Mercury chloride in our spheroids indicate charge selectivity through the barrier. Further analyses of tight junction distribution showed disorganization of junctional proteins for organoids cultured under hypoxic condition, pro-inflammatory cytokine levels were elevated under hypoxic condition, and BBB permeability was increased in hypoxic organoids. This in vitro system would have applications in drug discovery and neurotoxicity testing. In addition, through the use of representative cell types derived from induced pluripotent stem cells (iPSCs), individualized, patient-specific blood brain barrier disease models may be feasible.

 

Presenter contact: nzoug@pennmedicine.upenn.edu

 

Poster 12

 

A dual-mechanism antibiotic targets Gram-negative bacteria and avoids drug resistance

 

J.K. Martin1, M.Z. Wilson1,5, G.M. Moore1, J.P. Sheehan1, A. Mateus4, S.H.J. Li1, B.P. Bratton1,2, H. Kim3, J.D. Rabinowitz2,3, A. Typas4, M.M. Savitski4, Z. Gitai1

 

1 Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA

2 Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ 08544, USA

3 Department of Chemistry, Princeton University, Princeton, NJ 08544, USA

4 Genome Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany
5 University of California--Santa Barbara, Santa Barbara, CA, USA

 

The rise of antibiotic resistance and declining discovery of new antibiotics have created a global health crisis. Of particular concern, no new antibiotic classes have been approved for treating Gram-negative pathogens in decades. Here, we characterize a compound, SCH-79797, that kills both Gram-negative and Gram-positive bacteria through a unique dual-targeting mechanism of action (MoA) with undetectably low resistance frequencies. To characterize the MoA of SCH-79797 we combined quantitative imaging, proteomic, genetic, metabolomic and cell-based assays. This pipeline shows that SCH-79797 has two independent cellular targets, folate metabolism and bacterial membrane integrity, and outperforms combination treatments with other antifolates and membrane disruptors in killing MRSA persister cells. In an effort to both better understand its mechanism of action and improve its efficacy, we designed and synthesized a small library of SCH-79797 derivatives. These derivatives showed that the anti-folate and membrane-targeting activities of SCH-79797 are chemically separable. Furthermore, one of the derivatives led to a ~100-fold increase in antibiotic efficacy towards drug-resistant pathogenic bacteria, such as Neisseria gonorrhoeae. We are currently in the process of assessing the toxicity and in vivo efficacy of our compounds in rodent infection models. Thus, SCH-79797 and its derivatives represent promising lead antibiotics. We also suggest that combining multiple MoAs onto a single chemical scaffold may be an underappreciated approach for targeting challenging bacterial pathogens.

 

Presenter contact: jpsheehan@princeton.edu

 

Poster 13

 

Teaching the pharmaceutical research, approval and regulatory processes related to drug discovery

 

Doreen Badheka1 and Nicholas Ponzio1,2

 

1 Rutgers School of Graduate Studies, Newark Health Science Campus, 185 South Orange Avenue, Newark, NJ 07103

2 Department of Pathology, Immunology and Laboratory Medicine, Rutgers New Jersey Medical School, 185 South Orange Avenue, Newark, NJ 07103

 

Pharmacology courses and biomedical research are part of the curriculum of most health professional schools, but there are few opportunities for students to learn the entire process of how new drugs/treatments are conceived, developed, manufactured and brought to market. Our Molecules to Medicines (M2M) course is designed to provide such knowledge using novel educational strategies that expose students to each stage of the drug development process, and also provides an opportunity to work as a member of a virtual drug discovery Project Team. Rutgers Biomedical and Health Sciences (RBHS) is comprised of 8 health related schools, including medical, dental, nursing, public health and other health-related professions, including Rutgers School of Graduate Studies (SGS). The M2M course is offered through the Rutgers SGS as a 3-credit elective, but students from other RBHS programs can also cross register for the course. In addition to learning how drugs are developed, M2M also provides: (1) insight to activities that are common in industry, (2) an opportunity to understand what a career in a pharmaceutical company entails and (3) interaction with professionals from the pharmaceutical industry who give the lectures and provide guidance throughout the course. M2M is taught by an interdisciplinary faculty drawn from academe and industry. The most important elements of the teaching methods are having students work as part of a team multiple projects and presentations, as well as the consistent guidance given to each team by a core of co-course directors.

 

Presenter contact: doreen.badheka@rutgers.edu

 

Poster 14

 

The Selective Group I Metabotropic Glutamate Receptor Agonist 2-Chloro-5-hydroxyphenylglycine (CHPG) Enhances BDNF and Reverses Demyelination

 

Kyle S. Saitta1,2, Lauren D. Lercher2, Ashish Patel2, Yangyang Huang2, W. Geoff McAuliffe2, Cheryl F. Dreyfus2

 

1 Joint Graduate Program in Toxicology, Rutgers University, Piscataway, NJ 08854

2 Neuroscience and Cell Biology, Rutgers Robert Wood Johnson Medical School, Piscataway, NJ 08854

 

The demyelinating agent cuprizone (0.2%) causes a decrease in brain-derived neurotrophic factor (BDNF) and myelin proteins with an upregulation of metabotropic glutamate receptors (mGluRs) within the lesion site. Similarly, patients with multiple sclerosis not only have deficits in myelin, but also have enhanced expression of mGluRs within active chronic lesions and decreases in BDNF. Therefore, an intriguing therapeutic approach for these types of demyelinating diseases may be to enhance endogenous sources of BDNF. The aim of this study was to enhance BDNF through the application of 2-chloro-5-hydroxyphenylglycine (CHPG), a Group I mGluR agonist, by using the clinically relevant approach of a peripheral injection. Cuprizone or control feed was fed to adult mice prior to intraperitoneal (ip) injections of saline vehicle or CHPG (20 or 40 mg/kg). CHPG increases levels of BDNF and myelin proteins 24 hours after injection without affecting these proteins in control-fed mice. Furthermore, BDNF and myelin proteins remain elevated following administration of CHPG every other day for 2 weeks. The same dosing regimen of CHPG also increases myelin thickness and numbers of myelinated fibers following cuprizone as analyzed by transmission electron microscopy and improves behavioral deficits as analyzed by the balance beam assay and wire hang test. To elucidate the responsible receptor and site of action, the mGluR5 antagonist, 2methyl-6- (phenylethynyl)pyridine (MPEP), was injected into the lesion, followed by ip injection of CHPG. MPEP blocked effects of CHPG, suggesting that mGluR5 mediates actions of CHPG and that CHPG acts within the lesion site. Taken together, these data suggest that selective mGluR Group I agonists such as CHPG may be a therapeutic approach for treating demyelinating diseases by increasing the levels of BDNF and improving myelination. (Supp: NIH NS036647; T32ES007148; F31NS098642 and NMSS RG 4257B4/1)

 

Presenter contact: kyle.saitta@rutgers.edu

 

Poster 15

 

Engineered EF-Tu and tRNA in FRET-based screening assay for identification of inhibitors of protein synthesis in bacteria

 

Maxim Chudaev, Rachana Bhatt*, Wlodek Mandecki and Emanuel Goldman

 

Department of Microbiology, Biochemistry & Molecular Genetics, Rutgers-New Jersey Medical School, 225 Warren St., Newark, NJ 07101

* Current address: Department of Biomedical Engineering, Rutgers University, 599 Taylor Road Piscataway, NJ 08854

 

Antibiotic-resistant infections that do not respond to available drugs are becoming more common. Identifying EF-Tu targeting antibiotics is of significance especially that the EF-Tu protein sequence is highly conserved among bacteria and sufficiently different from both eukaryotic homolog eEF1A and human mitochondrial EF-Tu (1), a condition for the antibacterial drug’s specificity. The principle of the assay is single-pair fluorescence resonance energy transfer involving two molecules, a specific aminoacyl-tRNA (aa-tRNA) conjugated to a Cy3 fluorescent dye (donor) and the engineered elongation factor Tu (EF-Tu) conjugated to Cy5 (acceptor). The engineered EF-Tu contains three mutations that facilitate primarily site-specific attachment of a fluorophore while allowing the molecule to retain full biologic function in every assay in which we have tested it (2). Formation of the ternary complex (EF-Tu:tRNA:GTP) is targeted. When the reagents are mixed and Cy3 is excited at 532 nm, an increased Cy5 fluorescence intensity is observed at 665 nm due to ternary complex formation and FRET (3, 4). In mini-screening experiment on 96-well microplate with twenty different antibiotics, including β-lactams, quinolones, and various protein synthesis inhibitors, we observed that these antibiotics did not significantly affect the FRET (1). If the same assay is carried out in presence of a known inhibitor of the EF-Tu:tRNA interaction, such as GE2270A (1), kirromycin or factumycin (unpublished), the fluorescence intensity is significantly diminished. The work is ongoing towards adapting the assay to a homogeneous HTS format.

 

1. Bhatt R, Chudaev M, Mandecki W, Goldman E (2018) Engineered EF-Tu and tRNA-Based FRET Screening Assay to Find Inhibitors of Protein Synthesis in Bacteria Assay Drug Dev Technol.;16(4):212-221 PMID: 29870274

2. Maxim Chudaev, Kiran Poruri, Emanuel Goldman, Hieronim Jakubowski, Mohit Raja Jain, Wei Chen, Hong Li, Sanjay Tyagi, and Wlodek Mandecki. Design of the efficient tRNA:EF-Tu system for single molecule FRET studies of ribosomal translation. Protein Eng. Des Sel. Feb 26 (2013). PMID:23447652

3. Mandecki W, Bharill S, Borejdo J, et al. (2008) Fluorescence enhancement on silver nanostructures: studies of components of ribosomal translation in vitro. In: Single Molecule Spectroscopy and Imaging (eds. Enderlein J, et al.) Proc. of SPIE, vol. 6862: 68620T.

4. Perla-Kajan J, Lin X, Cooperman B, Goldman E, Jakubowski H, Knudsen C, Mandecki W (2010) Properties of E. coli EF-Tu Mutants Designed for Fluorescence Resonance Energy Transfer from tRNA Molecules. Protein Eng, Design & Sel. doi: 10.1093/protein/gzp079

 

Presenter contact: chudaymv@rutgers.edu

 

Poster 16

 

Targeting carbonic anhydrase enzymes to prevent mast cell development and mastocytosis

 

Christina M. Hernandez, Juan M. Inclan-Rico, and Mark C. Siracusa

 

Center for Immunity and Inflammation, Department of Medicine, New Jersey Medical School, Rutgers-The State University of New Jersey, 205 South Orange Ave. Newark, New Jersey, 07103.

NemaGen Discoveries, 155 Village Blvd. #130 Princeton, New Jersey, 08540

 

Mastocytosis is a rare disorder characterized by increased mast cell development and the accumulation of mast cells in various tissues. When properly regulated, small numbers of mast cells line barrier surfaces such as the skin and gut where they are activated in responses to pathogens. However, in the context of mastocytosis, the presence and activation of mast cells is dramatically increased; resulting in the release of histamines, leukotrienes, prostaglandins and other molecules that promote the development of hives, anemia, abdominal pain, organ damage and life-threatening anaphylaxis. Although patients suffering from mastocytosis are prescribed antihistamines and mast cell-stabilizers, treatment options are limited, and more severe forms of the disease remain life-threatening. Therefore, novel treatment strategies to inhibit mast cell development and activation are urgently needed. Our studies have revealed that the enzyme carbonic anhydrase (Car)1 is exclusively expressed by mast cell progenitors. Further, we have demonstrated that inhibition of Car1 is sufficient to prevent murine and human mast cell development in vitro as well as mast cell responses in a murine model of mastocytosis. Our current work seeks to advance our studies by developing more potent Car1 inhibitors capable of preventing mastocytosis at concentrations that would be ideal for translational consideration. To support these goals, we have developed a systemic approach that allows us to efficiently screen novel inhibitors via enzymatic, cell-based and in vivo assays. To date, we have identified several lead compounds that dramatically outcompete known Car enzyme inhibitors and can be taken forward for pharmacokinetic analysis.

 

Presenter contact: mark.siracusa@rutgers.edu

 

Poster 17

 

Using CCR1 Antagonists to Target Tumor Associated Macrophages

 

Salvatore Coniglio, Gregory Marshall, Mikhail Zagurskiy, Madeline Spinelli, Patrick Gabriel, Yuriko Root, James R. Merritt   

 

New Jersey Center for Science, Technology & Mathematics (NJCSTM), Kean University, STEM 518B, 1000 Morris Ave. Union, NJ 07083

 

Glioblastoma Multiforme (GBM) is the most aggressive form of adult brain tumor with a median survival time of twelve months. GBM is highly resistant to conventional therapy which includes surgical resection of the tumor, radiation treatment and chemotherapy (list agents).  GBM cells are highly motile and invasive resulting in infiltrative tumors with poorly defined borders.  GBM tumors are heavily infiltrated with microglia cells which are known to stimulate GBM cell invasion.  Furthermore, compounds that inhibit microglia, such as pexidartinib (PLX3397) can inhibit GBM invasion in vivo. A variety of chemokines are upregulated in the GBM tumor microenvironment and facilitate “cross-talk” between microglia and GBM cells eliciting a chemotactic response.  We have demonstrated that the chemotactic ligand, CCL3, is similarly upregulated in GBM tumors.  We postulated that inhibition of CCL3 associated receptors such as C-C receptor 1 (CCR1) might also inhibit GBM invasion, thus, a CCR1 antagonist could prove efficacious for blockade of microglia-induced glioblastoma invasion in vitro. Many potent CCR1 antagonists have been described in the literature.  We chose four of these compounds with two distinct structural cores, all with reported IC50’s of less than 200 nM for inhibition of CCR1 binding versus CCL3.  We examined the ability of these antagonists to block microglia-stimulated glioblastoma invasion using an in-vitro coculture invasion assay.

 

Presenter contact: coniglsa@kean.edu

 

Poster 18

Research Pathology Services

 

M. Polunas, P. Louro, R. Corrales, C. Walinsky, M. Goedken

 

Rutgers University Biologic Sciences Core Facilities, 41 Gordon Road Suite B, Livingston Campus, Piscataway NJ 08854

 

The mission of Research Pathology Services is to support research in animal models of human disease by providing histology techniques and veterinary pathology services. With expertise in a wide range of mammalian and non-mammalian species, Research Pathology Services assists investigators with the selection of animal models, design of experiments, analysis of gross and microscopic tissues, and interpretation of findings. Research Pathology Services is equipped with state-of-the-art equipment and is staffed by an ACVP boarded veterinary pathologist and three full-time technicians. Path Services encourages early partnership with investigators during the development of research projects in order to provide the best research, technical, and administrative support. As a component of the Office Research and Economic Development (ORED), Path Services supports academic investigators at Rutgers University, NJ county medical examiners offices, local biotech companies and external academic institutions.

 

Presenter contact: michael.goedken@rutgers.edu

 

Poster 19

 

L-Cystine Diamides as a Novel Therapy for Cystinuria

 

Longqin Hu1 and Amrik Sahota2

 

1 Department of Medicinal Chemistry, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, Piscataway, New Jersey 08854

2 Department of Genetics, Rutgers, The State University of New Jersey, Piscataway, New Jersey 08854

 

Cystinuria is a genetic disorder involving the abnormal transport of L-cystine from the renal proximal tubule and intestines leading to formation of L-cystine kidney stones in affected patients. In a collaborative effort, we are developing L-cystine diamides in a novel approach to prevent stone formation in cystinuria patients through the binding of tailored crystal growth inhibitors to L-cystine crystal surfaces. L-Cystine bismorpholide (LH707) and L-cystine bis(N'-methylpiperazide) (LH708) are two more stable and more effective L-cystine analogs that were discovered at Rutgers to overcome the druggability problems of L-cystine dimethyl esters (CDME), that was first reported to inhibit L-cystine crystal growth. These L-cystine diamides were shown to effectively inhibit L-cystine crystallization using real-time in situ atomic force microscopy and be orally bioavailable and efficacious in a knockout mouse model of cystinuria. The IP is currently licensed to PharmaKrysto (Scotland, UK).

 

Presenter contact: longhu@pharmacy.rutgers.edu

 

Poster 20

Molecular Design and Synthesis Group Collaborations, Rutgers University Biomedical Research Innovation Cores (RUBRIC)

 

Jacques Y. Roberge

 

Molecular Design and Synthesis, Rutgers University Biomedical Research Innovation Cores, Rutgers University, Wright-Rieman Building, 610 Taylor Rd, Piscataway, NJ 08854.

 

The Molecular Design and Synthesis Laboratory focuses on the design and synthesis of small molecule probes to investigate new biology. In collaboration with other biomedical scientists, these probes are used to interrogate biological pathways to create preliminary data for grant applications as well as to design molecules for diagnostic and therapeutic use. The Molecular Design and Synthesis core has established research collaborations with biotech companies as well as with internal Rutgers University investigators in multiple departments including RBHS and CINJ and is housed in fully equipped laboratories in the Chemistry department in Rieman Hall.

 

Presenter contact: jr1257@ored.rutgers.edu

 

Poster 21

Isocotoin potently inhibits hepatitis E virus replication through interference with heat shock-protein 90

 

Ila Nimgaonkar1, Nicholas F. Archer1, Isabelle Becher2, Mohammad Shahrad3, Andre Mateus2, Qiang Ding1, Florian Douam1, Jenna M. Gaska1, Mikhail M. Savitski2, Hahn Kim4, Alexander Ploss1

 

1 Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA

2 Genome Biology Unit, European Molecular Biology Laboratory, 69117 Heidelberg, Germany

3 Department of Electrical Engineering, Princeton University, Princeton, NJ 08544, USA

4 Department of Chemistry, Princeton University, Princeton NJ 08544, USA

 

There is no direct-acting antiviral treatment for hepatitis E virus (HEV), which causes ~3 million symptomatic infections and ~70,000 deaths per year globally. Following infection, immunocompromised persons and pregnant women experience particularly severe clinical manifestations including liver cirrhosis and acute liver failure respectively. Ribavirin monotherapy can be used to treat chronic hepatitis E in solid-organ transplant recipients, however it is not safe for pregnant women and furthermore ribavirin-resistant HEV strains are emerging in Europe. Therefore, novel therapies against HEV are urgently needed. Using a high-throughput screening assay, we identified isocotoin, a spatially-oriented hydroxy bi-aryl compound that effectively inhibits HEV replication. Isocotoin has low cytotoxicity and is effective against HEV strains from different genotypes. A cellular thermal shift assay determined that isocotoin binds to heat shock protein 90 (HSP90), a host protein not previously known to play a role in HEV replication. Specific inhibition or knockdown of HSP90 confirmed that it is an essential host factor to the HEV life cycle. Therefore HSP90 inhibitors, which are currently in clinical development as anti-cancer agents, may be promising candidates for novel direct-acting therapies against HEV.

 

Presenter contact: inimgaon@princeton.edu

 

Poster 22

Translation of Nanoparticle Therapeutics from Laboratory Discovery to Clinical Application: Scaled-up production and processing

 

Chang Tian1, Jie Feng2, Chester E. Markwalter1, Leon Z. Wang1, Madeleine Armstrong1, Robert K. Prud’homme1

 

1 Department of Chemical and Biological Engineering, Princeton University, Princeton, New Jersey, 08540

2 Department of Mechanical Science and Engineering, University of Illinois at Urbana-Champaign, Urbana, Illinois, 61801

 

Malaria is a major cause of mortality in developing tropical regions and its effective low-cost therapies are in much need. Lumefantrine is a Biopharmaceutics Classification System class II drug and can be used as a cure for malaria. We have developed in the past low-cost lumefantrine encapsulated nanoparticles (NPs) using stabilizers such as zein/casein and HPMCAS via Flash Nanoprecipitation (FNP) method. A continuous production procedure of NPs is set up to transfer the nanoparticle production from laboratory scale to clinical level. Using mixers of three different scales, the production rate of NPs is increased from a batch of a few milligrams to a continuous production of a hundred kilograms per day. An ultrafiltration system has been constructed to increase the NP concentration after FNP, which greatly improves the NP solidification efficiency. The pH of NP suspension, shear rate and transmembrane pressure (TMP) of NP flow within different types of filters are carefully controlled to prevent NP aggregation and filter cake accumulation. Next, scalable and continuous spray drying was applied to NP suspension of increased concentration to obtain dried powders with long-term storage stability. The dissolution kinetics of dried NP powders are showed to be significantly improved compared to that of crystalline lumefantrine in simulated fasted and fed intestine media. These results demonstrate complete translation of therapeutic nanoparticles from laboratory discovery to large scale clinical production.

 

Presenter contact: ctian@princeton.edu

 

Poster 23

Protein and Peptide Formulations in Highly-Loaded Nanocarriers

 

C.E. Markwalter1, R.F. Pagels1, A.N. Hejazi1, A.G.R. Gordon1, A.L. Thompson1, R.K. Prud'homme1

 

1 Department of Chemical and Biological Engineering, Princeton University, 44 Olden St Princeton, NJ 08542

 

The encapsulation of water-soluble therapeutics and biologics into nanocarriers to achieve novel therapeutic behavior has been envisioned for decades, but clinical translation has been hampered by complex production strategies. The methods that have been developed are also limited by poor encapsulation efficiency/loading and biologic instability. To address this unmet need, we introduce a solubility-driven self-assembly process to form polymeric nanocarriers comprising a biologic in a hydrophilic core, encapsulated by a poly(lactic acid) shell, and stabilized by a poly(ethylene glycol) brush. Called “inverse Flash NanoPrecipitation”, the technique achieves biologic loadings (w% of total formulation) that are 5-15x higher than typical values (9-27% versus <2%). In contrast to liposomes and polymersomes, we sequentially assemble the polymer layers to form the final nanocarrier. Installation of the poly(lactic acid) shell before water exposure sequesters the biologic in the core and results in the anomalously high loadings that are achieved. We have demonstrated the broad applicability of the process by formulating over a dozen different oligosaccharides, antibiotics, peptides, proteins, and RNA into nanocarriers with narrow size distributions and high reproducibility. Lysozyme and horseradish peroxidase are shown to release from the nanocarriers with 99% retained activity. These results demonstrate the potential for commercial implementation of this technology, enabling the translation of novel treatments in immunology, oncology, or enzyme therapies.

 

Presenter contact: cem3@princeton.edu

 

Poster 24

Reactivation of mutant p53 by small molecules for potential cancer therapeutics

 

M.R. Pardo1, S.L. Coverdale1, R. Sayed1, E. Peñate1, A. LaRocca1, B. Dasmahapatra1

 

1 Drew University, 36 Madison Ave., Madison, NJ 07940

 

Cancer is a major global public health issue, and the second leading cause of death in the United States. It is a group of complex diseases characterized by the uncontrolled growth of abnormal cells, often caused by the loss of genomic integrity in cells. Cellular genomic integrity is maintained by the tumor suppressor protein p53, which is also known as: the “guardian of the genome”. P53 prevents oncogenic transformation by halting the cell cycle upon cellular stress, and activating protective methods such as DNA repair or elimination of abnormal cells via apoptosis. Tumor suppressor activity of p53 is attenuated in almost all human cancers. In 50% of cancers, p53 is inactivated due to mutation. Most of these oncogenic mutations are missense mutations in the DNA binding domain of the p53 protein, resulting in the loss of DNA binding activity due to a conformational change at physiological temperature. Cancers with mutant p53 are aggressive and often resistant to therapy, making the mutant p53 an attractive target for drug discovery research. We have developed a cell-based reporter gene assay to screen compounds for their ability to restore transcriptional activity of mutant p53. Several small molecules of different chemical structures have been identified through this method. These molecules induce transcription of p21, a p53-regulated gene, in cancer cells with mutant p53.  Using conformation specific antibody, we have also shown that these molecules change mutant conformation to wild type conformation, underlying the mechanism of reactivation.

 

Presenter contact: mpardo@drew.edu

 

Poster 25

Discovery of PqsE Thioesterase Enzyme Inhibitors for the Bacterial Pathogen Pseudomonas aeruginosa using DNA-Encoded Small Molecule Library Screening

 

Julie S. Valastyan1,2, Michael Tota3, Isabelle R. Taylor1, Vasiliki Stergioula1, Chari D. Smith1, Brad R. Henke4, Graham A.B. Hone3, Kenneth G. Carson3, Bonnie L. Bassler1,2

 

1 Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA.

2 Howard Hughes Medical Institute, Chevy Chase, MD 20815, USA.

3 Macroceutics Incorporated, Monmouth Junction, NJ 08852, USA.

4 Opti-Mol Consulting, LLC, Cary, North Carolina 27513, USA.

 

Pseudomonas aeruginosa is an opportunistic bacterial pathogen responsible for a high incidence of life-threatening hospital-acquired infections. P. aeruginosa pathogenicity relies on the process of bacterial cell-cell communication known as quorum sensing. Quorum sensing allows bacteria to count the members of the vicinal community and undertake collective behaviors, such as biofilm formation and virulence factor production, only when there are enough cells present for group behaviors to effectively promote survival and/or infection. A thioesterase, PqsE, plays a vital role in quorum sensing and is absolutely essential for virulence in mouse and nematode models of infection. We used a DNA-encoded small molecule library screening strategy to discover new molecules that bind PqsE, and further validated select hits as non-competitive enzymatic inhibitors. The hit molecules were re-synthesized, free of their DNA barcodes, and confirmed for their inhibitory activity, and preliminary structure-activity relationships were determined through a series of analogs. The molecules identified in this screen serve as starting points for the development of potentially highly effective antibiotics for a deadly pathogen, as well as tool compounds in the elucidation of how PqsE drives quorum sensing phenotypes in P. aeruginosa.

 

Presenter contact: juliev@princeton.edu, itaylor@princeton.edu

 

Poster 26

Cell and Cell Products Fermentation Facility

 

Arvin Lagda

 

Waksman Institute of Microbiology, 190 Frelinghuysen Road, Piscataway, NJ 08854

 

The Cell and Cell Product Fermentation Facility at Waksman Institute has a multi-client base and produces a wide range of bulk biologics including: antimicrobials, cosmetic substrates, flavors/fragrances and plasmid-derived proteins, enzymes, growth factors and diagnostics to name just a few. We are equipped to handle most microbial fermentation requests with the exceptions of pathogenic or opportunistic organisms. Our bioreactor volumes range from 15 Liters to 800 Liters maximum operating volumes. All projects are conducted under CDA/NDA agreements, which limit discussion of specific company or projects. We operate as a not-for-profit, fee-for-service, self-supportive lab and receive no direct funding from state or federal sources.

 

The primary goal has been to provide affordable fermentation services to a highly diverse client base which includes but not limited to the academia, pharmaceutical and biotech industries who find their in-house facilities overtaxed.

 

Presenter contact: arvin.lagda@waksman.rutgers.edu



Poster 27

Selected Applications by NMR

 

István Pelczer and Kenith Conover

 

Department of Chemistry, Princeton University, Frick Chemistry Laboratory, Princeton, NJ 08544

 

Our NMR Facility is equipped with top of the line instruments between 10 MHz and 800 MHz, most of them are highly automated and several of them are equipped with cryoprobe.  This allows us also to run special experiments, which would not be available with more conventional hardware.  The poster highlights a few representative out-of-ordinary applications.

 

Our NMR Facility is open to all on campus, is available to academic users through collaboration, while outside industrial users can access it (and more) through the Industrial Associates Program (IAP) run by the Department of Chemistry (see flyer).

 

Presenter contact: ipelczer@princeton.edu

 

Poster 28

The Prism Imaging and Analysis Center − An engine for innovation and discovery

 

Nan Yao

 

Princeton University, 86 Olden Street, Princeton, NJ 08544

 

This poster will provide an introduction of Princeton's Imaging and Analysis Center (IAC) − An engine for innovation and discovery. Over the past 25 years, the IAC has developed to the largest central facility at Princeton and a world-leading center for both hard and soft materials (including biological specimens) characterization. IAC is a knowledge based materials characterization facility that provides teaching, expertise, collaboration, and instrumentation for advancing science and technology. IAC stands in the forefront[HH1]  of the innovation and discovery, and provides a unique platform and bridge for catalyzing and practicing collaborations at Princeton, and beyond. For more information, please contact Nan Yao, IAC Director, nyao@princeton.edu, or visit the center at Princeton or on the Web at iac.princeton.edu.

 

Presenter contact: nyao@princeton.edu

 

Poster 29

 

The Biological Mass Spectrometry Facility at CABM, Rutgers

 

Haiyan Zheng, Meiqian Qian, Caifeng Zhao, David Sleat and Peter Lobel

 

Rutgers University, 174 Frelinghuysen Rd, Piscataway, NJ 08854

 

The Biological Mass Spectrometry Facility at RWJ Medical School and Rutgers is a highly-regarded facility among the Rutgers research community and over the years, we have also established a broad customer base in other universities, non-profit organizations and the pharmaceutical/biotech industry. Our facility has cutting-edge mass spectrometry instrumentation (e.g., the Thermos Orbitrap Eclipse Tribrid and Thermo Q Exactive HF) and capable scientists with years of experience in protein-peptide mass spectrometry including protein complex analysis, short-gun proteomics, isobaric labeling proteomics, targeted analysis (PRM), PTM analysis on single proteins and at proteomic level. Other applications conducted by the Facility that are highly relevant to drug discovery include serum proteomics (targeted and non-targeted) and structural analysis of drug binding of target proteins using Hydrogen-deuterium exchange. Please visit our websites for details: http://cabm-ms.cabm.rutgers.edu/

 

Presenter contact: Haiyanz@cabm.rutgers.edu

 

Poster 30

Proteomics and Mass Spectrometry Core Facility

 

Saw Kyin

 

Department of Molecular Biology, Princeton University, LTL 016, Princeton, NJ 08544

 

We will present proteomics and small molecule analysis services and various mass spectrometry instruments available at Proteomics and Mass Spectrometry core facility of the Department of Molecular Biology at Princeton University.  Our facility provides full service as well as open access service at reduced rates.  Collaborated opportunity is also available.

 

Presenter contact: skyin@princeton.edu

 

Poster 31

Princeton University Flow Cytometry Resource Facility

 

Christina J. DeCoste and Katherine Rittenbach

 

Princeton University, Department of Molecular Biology, Flow Cytometry Resource Facility

 

The Flow Cytometry Resource Facility provides support for Princeton University faculty & student research and is available for outside clients on a fee-for-service basis. The facility is currently equipped with one analytical flow cytometer (BD Biosciences LSRII w/HTS system, 5-laser, 12-color), two cell sorters (BD Biosciences FACSAria Fusion sorter, 5-laser, 17-color; Bio-Rad S3e sorter, 2-laser, 4-color), one large-particle cell sorter (Union Biometrica COPAS system), a particle counter (Coulter Z2), and a cytospin centrifuge.  In addition to the instrumentation available, the Flow Cytometry Resource Facility also provides expert consultation and resources to assist with experimental design and troubleshooting, staining/sample preparation protocols, data acquisition, analysis and interpretation, and the preparation of publications and grant submissions. Training and education for independent use of the benchtop systems is provided regularly in one-on-one sessions. The journals Cytometry and Clinical Cytometry are available in the facility, as well as access to Current Protocols in Cytometry and relevant protocols and publications. On-site use of FACSDiVa, FlowJo and FCS Express software packages for data analysis is also available.

 

Presenter contact: cdecoste@princeton.edu

 

Poster 32

Nitrofurantoin tolerance of non-growing Escherichia coli and their re-sensitization

 

Sandra J. Aedo and Mark P. Brynildsen

 

Department of Chemical and Biological Engineering, Princeton University, 205 Hoyt Laboratory, 25 William Street, Princeton, NJ, 08544, USA

 

Nitrofurantoin is a broad-spectrum bactericidal antibiotic used in the treatment of urinary tract infections. It is a pro-drug that once activated by NfsA and/or NfsB goes on to inhibit bacterial DNA, RNA, and cell wall protein synthesis. Previously, it had been claimed that nitrofurantoin is active against both growing and non-growing bacteria. Here we have found that E. coli grown to stationary phase are completely tolerant to nitrofurantoin. Supplementation with glucose or gluconate under conditions where cells remained non-growing (different essential nutrients were absent) sensitized cultures to nitrofurantoin. We conceptualized nitrofurantoin sensitivity as a multi-input AND gate, and tolerance as an insufficiency in one or more of those inputs. The inputs considered were an activating enzyme, reducing equivalents, and cytoplasmic abundance of nitrofurantoin (because NfsA and NfsB are cytoplasmic proteins). We systematically assessed the contribution of each of these inputs to nitrofurantoin tolerance and found that the level of activating enzyme and nitrofurantoin import were not contributing factors to tolerance. Rather evidence suggested that the availability of reducing equivalents is why stationary phase E. coli are tolerant to nitrofurantoin and catabolites can re-sensitize those cells.

 

Presenter contact: saedo@princeton.edu

 

 


Poster 33

Application of the Clustering of Ligand Diffusion Coefficient Pairs (CoLD-CoP) Method to Detecting Cooperatively Binding Drug Fragments

 

S. V. Patnala1,2, D.A. Snyder1

 

1 William Paterson University, Chemistry Department, 300 Pompton Rd, Wayne, NJ

2 William Paterson University, Biology Department, 300 Pompton Rd, Wayne, NJ

 

Chemical screening, which accounts for 22% of biomedical R&D budgets, has a very high attrition rate. Improved experimental and computational techniques, as well as combinatorial paradigms such as fragment-based drug design, can mitigate ligand screening attrition risks. However, fragment-based drug design introduces a new issue: identifying which fragments bind cooperatively and hence, which fragments can be combined to produce a viable lead compound.

 

We extend the CoLD-CoP method, previously developed by Snyder and coworkers, to facilitate chemical screening to identify weakly binding ligands in the presence of nominal amounts of protein, to detect cooperative binding. Cooperatively binding fragments recognized by our method are puzzle pieces whose successive assembly is likely to yield a promising lead molecule. The approach can also identify competitive ligand binding, which is useful in probing wide-ranging enzyme active sites and protein chemistry.

 

Presenter contact: PatnalaV@student.wpunj.edu


Poster 34

 

Ploidy Impacts Persistence to Fluoroquinolones

 

Allison Murawski and Mark Brynildsen

Department of Molecular Biology, Princeton University, Princeton, NJ, USA

 

Non-growing bacterial populations are the most difficult to treat with antibiotics, and even for drugs that can kill the majority of growth-inhibited bacteria, such as fluoroquinolones (FQs), the presence of persisters can lead to treatment failure. While paradigms suggest that persisters survive due to limited antibiotic-induced damage, FQ persisters arising from stationary-phase populations experience significant FQ-induced DNA damage and require homologous recombination repair machinery to survive. Based on this knowledge, we hypothesized that ploidy present in stationary-phase Escherichia coli populations is a phenotypic variable that predisposes cells to become persisters. To assess this, we used the fluorescent, DNA specific dye Hoechst 33342 and sorted stationary-phase populations of E. coli based on chromosomal content. We found that subpopulations sorted to contain two or more chromosomes harbored greater than an order of magnitude more FQ persisters than those sorted to contain one chromosome. To assess the fidelity of the sorted populations with an independent approach, we used an origin reporter based on the parABS system and confocal fluorescence microscopy to quantify chromosome abundance. Those data confirmed that approximately 90% of cells in the one- and two-chromosome subpopulations were accurately sorted, and we used that data in conjunction with survival data to develop a mathematical model to quantify FQ persistence as a function of ploidy. The model suggested that few, if any, cells harboring one chromosome survive FQ treatment, which is consistent with homologous recombination as the major repair system used by persisters.  Further, we demonstrate that proteins involved in homologous recombination are required for this phenomenon, as the association between chromosome number and persistence was eliminated in a ΔrecB mutant. In addition, we show that our results are not unique to levofloxacin but extend to distinct fluoroquinolones, and that different environmental conditions or a clinical isolate of uropathogenic E. coli also exhibit a similar dependence of FQ persistence on ploidy. Together, our results identify chromosomal content as a major phenotypic variable that impacts whether cells will become FQ persisters, and we propose that the development of an FQ adjuvant that induces monoploidy could help to reduce the burden of chronic, relapsing infections.

 

Presenter contact: allisonm@princeton.edu