The Doctor of Philosophy Seminar and Examination of Jus Tupik

Biography

Juselyn (Jus) Tupik is a DVM/PhD dual degree student at the Virginia-Maryland College of Veterinary Medicine. Having lived in many states growing up, Jus has always called her grandparent’s house in the mountains of Pennsylvania home. Jus grew up alongside her twin brother and mom, who encouraged her childhood dream to become a veterinarian. As an undergraduate at the College of William and Mary, Jus conducted a research project on hemichordate worm ecology in the York River. It was through this research discovery process, from project design to data analysis, that sparked her interest in pursuing a Ph.D., in addition to veterinary school. Having a strong fascination with tick-borne and zoonotic diseases, Jus was thrilled to find her niche in host-pathogen relationships between the immune system and bacteria. In her opinion, immunology is one of the best fields to study themes in health and diseases, and bacteria are infinitely cooler than the other pathogens. 

As a dual degree student, Jus has had the opportunity to grow as a researcher and mentor. She served in multiple leadership positions, including co-president of the BMVS graduate student association and lead student coordinator of the National Association of Veterinary Scientists (NAVS). Jus also dedicated her time to the mentorship of 6 amazing research students, served as a science fair judge, and assisted pre-vet students with applying to vet school. Some of her favorite experiences at Virginia Tech include competing in the Nutshell games, going through LGBTQ+ SafeZone training, and learning about disability services for students abroad in the Global Perspectives Program. Jus’s favorite things about Blacksburg are the wonderful breweries, sunsets, and friendships. She is looking forward to finishing up the last few years of veterinary school and hoping to become a clinical microbiologist in the future. 

Funded by

National Institute of Health/National Institute of Allergy & Infectious Diseases: R03AI151494, R21AI159800

Virginia Tech/University of Maryland Joint Seed Grant

VMCVM Office of Research and Graduate Studies and DVM/PhD Student Support

Awards and Academic Achievements

·    2^nd place Oral Presentation: American Society of Microbiology Virginia Meeting 2023

·    Southeastern Immunology Symposium Young Investigator Award 2023

·    National DVM/PhD Combined Degree Colloquium Travel Award 2023

·    2^nd place Oral Presentation: American Society of Microbiology Virginia Meeting 2022

·    1^st Place Poster: VCOM Virginia Research Day 2022

·    American Association of Immunologists Abstract Trainee Award, 2021 & 2022

·    Phi Kappa Phi Honor Society Fall 2020

·    Phi Beta Kappa Inductee Spring 2019

Lay Language Abstract 

The immune system consists of two arms: a specialized adaptive response that forms long-lasting defense, and the generalized innate response that provokes immediate protection. Consequently, the innate immune system is our first line of defense against infectious diseases. It operates in part through proteins called Pattern Recognition Receptors (PRRs), which work by recognizing foreign signals in the body, known as Pathogen-Associated Molecular Patterns (PAMPs). The innate immune system’s job is to enforce a system of checks and balances to regulate inflammation in host defense. In this dissertation, we focus on one class of PRRs known as the NOD-like Receptors (NLRs). NLRs consist of pro-inflammatory receptors that form a multi-protein complex termed the inflammasome, as well as NLRs that instead regulate inflammation. Here, we investigated the role that inflammasomes and negative regulatory NLRX1 play in response to bacterial diseases.

First, we studied brucellosis, a chronic disease often transmitted in unpasteurized dairy products from livestock. Using mouse models and immune cells infected with Brucella abortus, we found that inflammasomes in murine models were protective against brucellosis through the initiation of inflammatory cell death. In contrast, the inhibition of inflammation by NLRX1 in mouse models adversely led to increased disease in the spleen and liver. Despite these contrasting results, Brucellagenomic DNA was an effective signal, or PAMP, for activation of both NLRs. These results suggest the importance of DNA recognition by NLRs, and that inflammation is protective during brucellosis. 

Second, we investigated NLRX1 regulation of Borrelia burgdorferi in Lyme arthritis. Characterized by persistent inflammation of the joints, Lyme arthritis is an enigmatic and difficult inflammatory condition to resolve. We found that in mouse models, NLRX1 was protective against arthritis. By analyzing changes in inflammatory gene and protein expression of infected ankle joints, followed by studies in immune and wound healing cells, we found that NLRX1 promotes faster cell migration and regulates cell metabolism. Our results suggest that NLRX1 may metabolically shift cells in the joint towards a more favorable wound healing environment for arthritis resolution. 

Ultimately, this work emphasizes the importance of balance in NLR signaling. Further, NLR signaling is highly context-specific for the cell type and bacterial disease, showcasing the complexity of host-pathogen interactions when battling bacteria.

Publications

Nagai-Singer, M.A.,Woolls, M.K., Leedy, K., Hendricks-Wenger, A., Brock, R.M., Coutermarsh-Ott, S., Paul, T., Morrison, H.A., Imran, K.M., Tupik, J.D., Fletcher, E.J., Brown, D.A., Allen, I.C. 2023. Cellular context dictates the suppression or augmentation of triple-negative mammary tumor metastasis by NLRX1. The Journal of Immunology, 211, 12: 1844-1857. 

Imran,K.M., Gannon, J., Morrison, H.A., Tupik, J.D., Tintera, B., Nagai-Singer, M.A., Ivester, H., Markov Madanick, J., Hendricks-Wenger A., Uh, K., Edwards, M., Coutermarsh-Ott, S., Eden, K., Byron, C., Clark-Deener, S., Lee, K., Vlaisavljevich, E., Allen, I.C.. 2023. Successful In Situ Targeting of Pancreatic Tumors in a Novel Orthotopic Porcine Model Using Histotripsy. Ultrasound Med Bio, 49,11.

Imran, K.M., Tintera, B., Morrison, H.A., Tupik, J.D., Nagai-Singer, M.A., Ivester, H., Council-Troche, M., Edwards, M., Coutermarsh-Ott, S.L., Byron, C.R., Clark-Deener, S., Uh, K., Lee, K., Boulos, P., Rowe, C., Coviello, C., Allen, I.C. 2023. Improved therapeutic delivery targeting clinically relevant orthotopic human pancreatic tumors engrafted in immmuocompromised pigs using ultrasound-induced cavitation: a pilot study. Pharmaceutics, Drug Delivery and Controlled Release, 15, 6, 1585.

Nagai-Singer, M.A., Morrison, H.A., Woolls, M.K., Leedy, K., Imran, K.M., Tupik, J.D., Allen, I.C. 2023. NLRX1 functions as a tumor suppressor in Pan02 pancreatic cancer cells. Frontiers in Oncology, Cancer Immunity and Immunotherapy, 13:1155831.

Tupik, J.D., Markov Madanick, J.W., Ivester, H.M., Allen, I.C. 2022. Detecting DNA: An Overview of DNA Recognition by Inflammasomes and Protection against Bacterial Respiratory Infections. Cells, 11, 1681.

Trusiano B., Tupik J.D., Allen I.C. 2022. Cold sensor, hot topic: TRPM8 plays a role in monocyte function and differentiation. J Leukoc Biol, 112, 3, 361-363.

Ivester, H.M., Tupik, J.D., Nagai-Singer, M.A., Coutermarsh-Ott, S.L., Allen, I.C. 2022. Methods to Evaluate Virus-Mediated Acute Lung Inflammation. Methods in Cell Biology, 168, 329-341.

Schrage, K. R., Tupik, J. D., & Allen, J. D. 2021. Intertidal zonation of hemichordates in soft sediments. Invertebrate Biology, 140, 3, e12344.

Tupik, J.D., Coutermarsh-Ott, S.L., Benton, A.H., King, K.A., Kiryluk, H.D., Caswell, C.C., Allen, I.C., 2020. ASC-Mediated Inflammation and Pyroptosis Attenuates Brucella abortus Pathogenesis Following the Recognition of gDNA. Pathogens, 9, 12.

Nagai-Singer, M.A., Hendricks-Wenger, Brock, R.M., Morrison, H.A., Tupik, J.D., Coutermarsh-Ott, S., Allen, I.C., 2020. Using Computer-based Image Analysis to Improve Quantification of Lung Metastasis in the 4T1 Breast Cancer Model. JoVE (164), e61805.

Tupik J.D., Nagai-Singer M.A., Allen I.C. 2020. To protect or adversely affect? The dichotomous role of the NLRP1 inflammasome in human disease. Mol Aspects Med, 76, 100858.

Presentations

*=national/international presentation, #=award winning

#Juselyn D. Tupik, Mecaila E. McClune, Julia A. Gregory, Jules M. Dressler, Brandon L. Jutras, Irving C. Allen. A “mighty” mitochondrial defense: Host immunoprotection by receptor NLRX1 during Lyme disease. Talk, American Society of Microbiology Virginia Chapter Meeting, November 2023, Richmond, VA.   

*#Juselyn D. Tupik, Mecaila E. McClune, Julia A. Gregory, Hailey W. Camp, Jules M. Dressler, Margaret A. Nagai-Singer, Brandon L Jutras, and Irving C. Allen. Mounting a mitochondrial defense: Metabolic immunoprotection by immune receptor NLRX1 during Lyme disease. Talk, National Combined DVM/PhD Degree Colloquium, August 2023, San Juan, Puerto Rico. 

#Juselyn D. Tupik, Mecaila E. McClune, Julia A. Gregory, Hailey W. Camp, Jules M. Dressler, Margaret A. Nagai-Singer, Brandon L Jutras, and Irving C. Allen. Mounting a Mitochondrial Defense: Immunometabolism and antimicrobial regulation by innate immune receptor NLRX1 during Lyme disease. Poster Presentation. Southeastern Immunology Symposium. June 2023, Nashville, TN. 

*Juselyn D. Tupik. Accommodating Students with Disabilities. Talk, Virginia Tech Steger Center GPP Global Seminar, May 2023, Riva San Vitale, Ticino, Switzerland. 

*Juselyn D. Tupik, Mecaila E. McClune, Hailey W. Camp, Julia A. Gregory, Jules M. Dressler, Justin W. Markov Madanick, Margaret A. Nagai-Singer, Brandon L Jutras, and Irving C. Allen. Upping the “anti-” for arthritis: Antimicrobial defenses through anti-inflammatory Nod-like Receptor NLRX1 during Lyme disease. Poster Presentation. American Association of Immunologists Annual Meeting, May 2023, Washington, D.C.

*Juselyn D. Tupik, Mecaila E. McClune, Hailey W. Camp, Julia A. Gregory, Jules M. Dressler, Justin W. Markov Madanick, Margaret A. Nagai-Singer, Brandon L Jutras, and Irving C. Allen. Host-pathogen interactions and antimicrobial defenses by innate immune receptor NLRX1 during Lyme borreliosis. Talk, International Symposium on Ticks and Tick-borne Diseases, March 2023, Virtual. 

Juselyn D. Tupik, Mecaila E. McClune, Hailey W. Camp, Julia A. Gregory, Jules M. Dressler, Justin W. Markov Madanick, Margaret A. Nagai-Singer, Brandon L Jutras, and Irving C. Allen. Mounting a mitochondrial defense: Innate immune receptor NLRX1 regulates cell metabolism and death for protection against Lyme disease. Poster Presentation. Biomedical & Veterinary Science Program Annual Research Symposium, March 2023, Blacksburg, VA.

Juselyn D. Tupik. Body vs Bacteria: How our immune system battles Lyme disease. Flash Talk, Virginia Tech Nutshell Games, February 2023, Blacksburg, VA.

Juselyn D. Tupik, Mecaila E. McClune, Hailey W. Camp, Julia A. Gregory, Jules M. Dressler, Justin W. Markov Madanick, Margaret A. Nagai-Singer, Brandon L Jutras, and Irving C. Allen. Upping the “anti” for arthritis: Antimicrobial defenses by anti-inflammatory receptor NLRX1 during Lyme disease. Poster Presentation. Edward Via College of Osteopathic Medicine Virginia Research Day, February 2023, Blacksburg, VA.

#Juselyn D. Tupik, Mecaila E. McClune, Jules M. Dressler, Justin W. Markov Madanick, Margaret A. Nagai-Singer, Hailey W. Camp, Brandon L Jutras, and Irving C. Allen. Battling Borrelia: The role of anti-inflammatory receptor NLRX1 in antimicrobial protection against Lyme disease. Talk, American Society of Microbiology Virginia Meeting, November 2022, Middletown, VA.

Juselyn D. Tupik, Justin W. Markov Madanick, Angela H. Benton, Kellie A. King, Hannah M. Ivester, Sheryl L. Coutermarsh-Ott, Clayton C. Caswell, and Irving C. Allen. Striking a balance: Investigating homeostasis in NOD-like Receptor (NLR) innate immune signaling during brucellosis. Poster Presentation, CeZAP Symposium, October 2022, Blacksburg, VA.

Juselyn D. Tupik, Mecaila E. McClune, Jules M. Dressler, Justin W. Markov Madanick, Margaret A. Nagai-Singer, Hailey W. Camp, Brandon L. Jutras, and Irving C. Allen. Attenuating Arthritis: The role of anti-inflammatory NLRX1 in protection against Lyme disease. Poster Presentation, Southeastern Immunology Symposium, June 2022, Durham, NC.

#*Juselyn D. Tupik, Angela H. Benton, Kellie A. King, Hannah M. Ivester, Justin W. Markov Madanik, Sheryl L. Coutermarsh-Ott, Clayton C. Caswell, and Irving C. Allen. The Goldilocks Conundrum: The protective and adverse roles of immunoregulation by NOD-like receptors (NLRs) during brucellosis. Talk & Poster, American Association of Immunologists Annual Meeting, May 2022, Portland, OR.

Juselyn D. Tupik, Mecaila E. McClune, Jules M. Dressler, Brandon L. Jutras, Irving C. Allen. Defense Against Disease: The protective role of anti-inflammatory receptor NLRX1 and its recognition of spirochete Borrelia burgdorferi during Lyme disease. Flash Talk, American Society of Microbiology DC Branch Meeting, April 2022, Virtual.

Juselyn D. Tupik, Mecaila E. McClune, Jules M. Dressler, Brandon L. Jutras, Irving C. Allen. A “sub-lyme” System: Determining the Protective Role of Anti-Inflammatory NIRX1 against Lyme Disease. Poster Presentation, 2022 ID IGEP Recruitment Event Virginia Tech, March 2022, Blacksburg, VA.

Juselyn D. Tupik, Mecaila E. McClune, Jules M. Dressler, Brandon L. Jutras, Irving C. Allen. Battling Bacteria: How Does Our Immune System Beat Lyme Disease?. Flash Talk, Virginia Tech Nutshell Games, March 2022, Blacksburg, VA.

Juselyn D. Tupik, Mecaila E. McClune, Jules M. Dressler, Brandon L. Jutras, Irving C. Allen. Detecting Disease: Defining the protective role of anti-inflammatory NLRX1 against Lyme disease. Poster Presentation, Virginia Tech Graduate Student Assembly Research Symposium, March 2022, Virtual.

#Juselyn D. Tupik, Mecaila E. McClune, Jules M. Dressler, Brandon L. Jutras, Irving C. Allen. A “sub-lyme” System: Determining the Protective Role of Anti-Inflammatory NIRX1 against Lyme Disease. Poster Presentation, Edward Via College of Osteopathic Medicine Virginia Research Day, February 2022, Blacksburg, VA.

Juselyn D. Tupik, Mecaila E. McClune, Jules M. Dressler, Brandon L. Jutras, Irving C. Allen. A ‘Sub-Lyme’ System: Uncovering mechanisms of NLRX1 immunoregulation of Borrelia burgdorferi during Lyme Disease. Talk, Biomedical & Veterinary Science Program Research in Progress Seminar, September 2021, Blacksburg, VA.

*Juselyn D. Tupik, Mecaila E. McClune, Jules M. Dressler, Brandon L. Jutras, Irving C. Allen. A “sub-Lyme” system: Uncovering the protective role of anti-inflammatory NLRX1 against Lyme disease. Talk & Poster, National Combined DVM/PhD Degree Colloquium, August 2021, Virtual.

#*Juselyn D. Tupik, Sheryl L. Coutermarsh-Ott, Kellie A. King, Angela H. Benton, Hanna D. Kiryluk, Clayton C. Caswell, Irving C. Allen. Till cell death do us part: The protective role of inflammasome-mediated pyroptosis and genomic DNA recognition during brucellosis. Talk & Poster, American Association of Immunologists Annual Meeting, May 2021, Virtual. 

Juselyn D. Tupik, Sheryl L. Coutermarsh-Ott, Kellie A. King, Angela H. Benton, Hanna D. Kiryluk, Clayton C. Caswell, Irving C. Allen. (Cell) Death by inflammasome: Uncovering mechanisms of host innate immune defense against the bacterial disease brucellosis. Flash Talk, Virginia Tech Graduate Student Assembly Research Symposium, March 2021, Virtual. 

Juselyn D. Tupik, Sheryl L. Coutermarsh-Ott, Kellie A. King, Angela H. Benton, Hanna D. Kiryluk, Clayton C. Caswell, Irving C. Allen. Brucella abortus genomic DNA enhances inflammasome recognition and facilitates protective inflammatory cell death during brucellosis. Talk, Biomedical & Veterinary Science Program Annual Research Symposium, March 2021, Virtual.

Juselyn D. Tupik, Sheryl L. Coutermarsh-Ott, Kellie A. King, Angela H. Benton, Hanna D. Kiryluk, Clayton C. Caswell, Irving C. Allen. Inflammasome-mediated inflammatory cell death protects against the zoonotic disease brucellosis. Poster Presentation, Edward Via College of Osteopathic Medicine Virginia Research Day, February 2021, Virtual. 

*Juselyn D. Tupik, Sheryl L. Coutermarsh-Ott, Kellie A. King, Angela H. Benton, Hanna D. Kiryluk, Clayton C. Caswell, Irving C. Allen. This means [inflammatory] war: Inflammasome recognition of the bacterium Brucella abortus. Poster Presentation, National Combined DVM/PhD Degree Colloquium, August 2020, Virtual. 

Juselyn D. Tupik, Sheryl L. Coutermarsh-Ott, Kellie A. King, Angela H. Benton, Hanna D. Kiryluk, Clayton C. Caswell, Irving C. Allen. This Means [Inflammatory] War: Targeting the deadly bacterium Brucella abortus. Flash Talk, Virginia Tech Graduate Student Assembly Research Symposium, April 2020, Virtual. 

Juselyn D. Tupik, Sheryl L. Coutermarsh-Ott, Kellie A. King, Angela H. Benton, Hanna D. Kiryluk, Clayton C. Caswell, Irving C. Allen. Brucella abortus is recognized by the inflammasome and negative regulator NLRX1 of the innate immune system. Poster Presentation, College of William and Mary Graduate Research Symposium, March 2020, Virtual.

Examination Graduate Committee

Major Advisor/Chair:

Irving Coy Allen, PhD, MBA
Professor of Inflammatory Diseases
Assistant Department Head for Research Support
Department of Biomedical Sciences & Pathobiology
Virginia-Maryland College of Veterinary Medicine (VMCVM), Virginia Tech

Graduate Advising Committee Members:

Brandon L. Jutras, PhD
Associate Professor
Department of Biochemistry, Virginia Tech
Department of Microbiology-Immunology, Northwestern University Feinberg School of Medicine

Christopher M. Reilly, PhD
Department Chair for Biomedical Sciences
Edward Via Virginia College of Osteopathic Medicine
Research Associate Professor
Virginia-Maryland College of Veterinary Medicine (VMCVM), Virginia Tech

Clayton C. Caswell, PhD
Associate Professor
Department of Biomedical Sciences & Pathobiology
Virginia-Maryland College of Veterinary Medicine (VMCVM), Virginia Tech

Sheryl L. Coutermarsh-Ott, DVM, PhD, DAVCP
Adjunct Professor
Department of Biomedical Sciences & Pathobiology
Virginia-Maryland College of Veterinary Medicine (VMCVM), Virginia Tech
Veterinary Pathologist, Virginia Department of Agriculture and Consumer Services

Yanjin Zhang, DVM, PhD
Associate Professor
Department of Veterinary Medicine

Virginia-Maryland College of Veterinary Medicine (VMCVM), University of Maryland