A. Personal Statement 

I am a basic scientist focused on the clinical and translational science with a track record of conducting research on drug delivery across the translational continuum. As the Associate professor at University of Pittsburgh, I have the expertise and motivation necessary to advance the imaging and care of BPH/LUTS in collaboration with Dr. Yoshimura, Chermansky and Sholosh. In this renewal application, we seek to advance the understanding of male LUTS by investigating cause and effect relationship of prostatic inflammation with molecular targeted inhibition of inflammation to empirically validate the replacement of Evans blue dye with Gadolinium based contrast agent in DCE-MRI of treated and untreated rodent groups modeling prostatic inflammation. Research on animal models will support DCE-MRI of BPH/LUTS patients and asymptomatic controls to tailor multiparametric MRI of BPH/LUTS by incorporating insights from completed U54 funded research and published basic research. Proposed translational research will benefit from my commitment and experience of taking discoveries from laboratory benchtop to the bedside, as exemplified by recent publications on basic and clinical research on novel contrast mixture for bladder MRI. Our translational research work on MRI has been recognized at international forums. This documented experience of research across the translational continuum from laboratory benchtop to bedside will be critical for the proposed project on BPH/LUTS. My PhD dissertation research on bladder instillation of liposomes for Interstitial cystitis (IC) was granted United States Patents: US 81,10217 (issued on 10-11-2006). Use of liposomes as delivery for Tacrolimus has now progressed from animal studies to Phase 2a clinical trial (NCT03129126) on hemorrhagic cystitis. My broad background in pharmacology, cancer drug delivery with specific training and expertise in key research areas for this application are directly relevant to the goals and activities of the current proposal. As our joint publications indicate, we have worked as a team in the past, which portends successful collaboration for this project: Singh N, Zabbarova I, Ikeda Y, Maranchie J, Chermansky C, Foley L, Hitchens TK, Yoshimura N, Kanai A, Kaufman J, Tyagi P (2020). Virtual measurements of paracellular permeability and chronic inflammation via color coded pixel-wise T(1) mapping. Am J Physiol Renal Physiol. 319(3):F506-F514. PMID:32715761 My work has been recognized by awards at NIH, AUA, society of female urology and urodynamics (SUFU) meetings and annual Jack Lapides essay contest. My major contributions to science, described in greater detail in section C. below, are directly relevant to the goals and activities of the current proposal.

1. Positions and Honors
   Positions and Employment

1996-1997         Analytical Chemist, Albert David Ltd. India
1998-1999         Research Assistant, Indian Council of Medical Research project, AIIMS, India
1999-2000         Senior Research Fellow, National Institute of Pharmaceutical Education
                          and Research, India
Summer 2001   Visiting Scholar, Surgery Department, National University of Singapore,
                          Singapore
2000-2005        Teaching Fellow, School of Pharmacy, University of Pittsburgh
2005-2006        Post-doctoral Fellow, Dept. of Urology, School of Medicine,
                         University of Pittsburgh
2006-2008        Research Assistant Professor, Dept. of Urology, School of Medicine,
                         University of Pittsburgh
2008- 2011       Director of Urology Laboratory, Beaumont Research Institute,
                         William Beaumont Hospital
2011-Present    Associate Professor, Dept. of Urology, University of Pittsburgh

Other Experience and Professional Memberships

2007-present    Affiliate member of American Urological Association

1997-present    Life Member of Indian Pharmacological Society

2007-present    Adhoc reviewer for multiple journals in urology and related fields:
                         Journal of Urology, Urology, Neurourology and Urodynamics, BJUI,
                         American J of Physiology, Spinal Cord, Scientific Reports, Journal
                         of Proteomics
2010-2011       Invited reviewer for the foundation of Wellbeing for Women, UK
2011-2012       Study section member for the VA Rehabilitation Research and
                        Development (RR&D) proposals
2012-2013       Prostate Cancer Research Program (PCRP) Idea Development Award
2014-2017       Member review panel for urology care foundation research proposals
2014                French National Research Agency ANR, External Reviewer
2015                Ad Hoc member for special emphasis panel ZRG1 EMNR
2016                Reviewer for Croatian Science Foundation
2017                Reviewer, Netherlands Organization for Scientific Research
2018                Reviewer, Wings of Life
2018                Adjunct chair, CSR special emphasis panel ZRG1 DKUS-R (12)
2018                Reviewer, CSR Anonymization Study (EMNR IRG)
2019                Reviewer for Paralyzed Veterans of America grants
2019                Peer Reviewed Medical Research Program of DOD

Honors

2002               Graduate student award for XIV World Congress of Pharmacology
                       at San Francisco
2003               First prize in Graduate student research excellence award School
                       of Pharmacy, University of Pittsburgh
2009              Winner of best clinical essay prize in SUFU meeting
2009              Winner of best basic science poster at the NIH, NIDDK meeting on
                      Urinary Incontinence
2009              2nd prize winner in the Jack Lapides essay contest
2010              Grand Prize Winner of Jack Lapides Essay Contest on Urodynamics
                      and Neuro-urology Research
2011              Best Paper award for research presented at OU-Beaumont Symposium
2011              Best basic science essay prize at SUFU for Dr. Vikas Tyagi (Mentee)
2012              Best Poster at the AUA on Prostate inflammation and Afferent Sensitization
2012              American Urological Association Foundation fellowship grant to Nirmal
                     Jayabalan (Mentee)
2013             American Urological Association Foundation fellowship grant to Mahendra
                     Kashyap (Mentee)
2015             Honorable mention in the Jack Lapides essay contest and best poster
                     award at AUA for radiation cystitis research
2015             Best poster at the AUA (4 in total) for work on pathogenic role of BDNF in OAB,
                     Inflammasomes in BPH, and NGF antisense treatment for autonomic dysreflexia.
2016             Best poster at the AUA for work on HCN channel function in human bladder
2017             Best poster at the AUA for microRNA-132 function in rat bladder
2018             Best poster at the AUA on novel contrast mixture for IC
2018             Best podium at the ICS on bladder wall imaging
2018             Best poster on BPH related detrusor underactivity, NIDDK conference

2. Contribution to Science selected from 96 research articles, 40 reviews

Magnetic resonance Imaging (MRI):  Over the last five years, we have leveraged our intravesical drug delivery expertise to advance MRI based artifact-free visualization of the thin bladder wall in rodents and in humans. Instillation of Gadobutrol and Ferumoxytol mixture permits mapping of T1 relaxation time of bladder wall for virtual histology and measurement of permeability in mouse bladder at 9.4T and human bladder at 3T. Nearly half-decade experience in MRI research will be critical for the success of proposed translation research on advancing the understanding of BPH/LUTS across the translational continuum.

1. Tyagi P, Janicki J, Hitchens K, Foley L, Kashyap M, Yoshimura N, Kaufman J (2017). Novel Contrast Mixture Improves Bladder Wall Contrast For Visualizing Bladder Injury. Am J Physiol Renal Physiol. 313(2): F155-F162. PMC5582910

2. Tyagi P, Janicki J, Moon CH, Kaufman J, Chermansky C (2018). Novel contrast mixture achieves contrast resolution of human bladder wall suitable for T1 mapping: applications in interstitial cystitis and beyond. Int Urol Nephrol. 50(3):401-409. PMC6028942

3. Tyagi P, Moon CH, Janicki J, Kaufman J, Chancellor M, Yoshimura N, Chermansky C (2018). Recent advances in imaging and understanding interstitial cystitis. F1000Res 2018, 7. PMC6234747

4. Singh N, Zabbarova I, Ikeda Y, Maranchie J, Chermansky C, Foley L, Hitchens TK, Yoshimura N, Kanai A, Kaufman J, Tyagi P (2020). Virtual measurements of paracellular permeability and chronic inflammation via color coded pixel-wise T(1) mapping. Am J Physiol Renal Physiol. 319(3):F506-F514. PMID:32715761

Prostate Inflammation:  Previously, we characterized the role of inflammasome and IL-1 in rat model of prostatic inflammation. To our surprise, IL-1 is not detectable in urine of BPH patients, but copious amounts of secretory IL-1 receptor antagonist (sIL-1Ra) is detectable in urine of BPH patients. IL-1Ra is an anti-inflammatory cytokine codified by IL1RN gene that blocks IL-1 signaling by acting like a drug antagonist, binds to IL-1 receptors without inducing any intracellular response. Recently we noted that lower urinary tract symptoms in rat model of prostatic inflammation are mechanistically linked to the phenotypic changes in function of innervated neurons. This demonstrated attention to biological effects of signaling mediators across a translational continuum will contribute to the success of project.

5. Kashyap M, Pore S, Gingrich J, Wang Z, Yoshimura N, Tyagi P (2015). Inflammasomes are important mediators of prostatic inflammation associated with BPH. J Inflamm. 17; 12:37. PMC4436794

6. Tyagi P, Motley SS, Koyama T, Kashyap M, Gingrich J, Yoshimura N, Fowke JH (2018). Molecular correlates in urine for the obesity and prostatic inflammation of BPH/LUTS patients. Prostate;78(1):17- 24.PMC5716834

7. Funahashi Y, Takahashi R, Mizoguchi S, Suzuki T, Takaoka E, Ni J, Wang Z, DeFranco DB, de Groat WC, Tyagi P, Yoshimura N (2019). Bladder overactivity and afferent hyperexcitability induced by prostate-to-bladder cross-sensitization in rats with prostatic inflammation. J Physiol. 597(7):2063- 2078 PMC6441933

8. Ni J, Mizoguchi S, Bernardi K, Suzuki T, Kurobe M, Takaoka E, Wang Z, DeFranco DB, Tyagi P, Gu B, Yoshimura N (2019). Long-lasting bladder overactivity and bladder afferent hyperexcitability in rats with chemically-induced prostatic inflammation. Prostate. 79(8):872-879. PMC7327236

Intravesical liposome-based drug delivery:  I lead the investigations into the mechanism by which liposomes are taken up by bladder and performed pharmaceutical and pharmacological studies on empty liposomes and liposomes loaded with tacrolimus to treat bladder wall inflammation. My basic research on empty liposomes led to the licensing of technology by the University of Pittsburgh, which is being tested in IC/PBS patients and results of pilot studies were published in J Urol. 2009 Oct;182(4):1393-400 and Int Urol Nephrol. 46(12):2291-5. Two decades long track record of conducting research across the translational continuum from laboratory benchtop to bedside will be valuable to the current proposal.

9. Nirmal J, Tyagi P, Chancellor MB, Kaufman J, Anthony M, Chancellor DD, Chen YT, Chuang YC (2013). Development of potential orphan drug therapy of intravesical liposomal tacrolimus for hemorrhagic cystitis due to increased local drug exposure. J Urol. 189(4):1553-8. PMID: 23127767

10. Nirmal J, Wolf-Johnston AS, Chancellor MB, Tyagi P, Anthony M, Kaufman J, Birder LA (2014). Liposomal inhibition of acrolein-induced injury in rat cultured urothelial cells. Int Urol Nephrol. 46(10):1947-52. PMC4177265

11. Bharathi Raja R, Chancellor MB, Nirmal J, Dang L, Tyagi P (2015). Bladder Uptake of Liposomes after Intravesical Administration Occurs by Endocytosis. PLoS One.;10(3):e0122766. PMC4374861

12. Bharathi Raja R, Janicki JJ, Levanovich P, Tyagi P, Hafron J, Chancellor MB, Krueger S, Marples B (2015). Intravesical Liposomal Tacrolimus Protects against Radiation Cystitis Induced by 3-Beam Targeted Bladder Radiation. J Urol. 2015. pii: S0022-5347(15)03648-4. PMC4509800 (paper won best poster award at AUA 2015)

Gene silencing and overexpression:  We used advanced drug delivery approaches with antisense oligo nucleotides and peptide nucleic acids to establish the pathogenic role of nerve growth factor (NGF) in bladder overactivity and afferent excitability. Our studies demonstrated that cationic peptides and lipids are critical for enhancing the bladder uptake of antisense oligos and deriving their therapeutic potential in voiding dysfunctions. This demonstrated attention to biological effects of single molecular entity will contribute to the success of the project.

13. Tyagi P, Banerjee RK, Basu S, Yoshimura N, Chancellor MB, Huang L (2006). Intravesical Antisense Therapy For Cystitis Using TAT-Peptide Nucleic Acid Conjugates. Mol. Pharm. 3(4):398-40. PMID: 16889433

14. Kashyap, M., Kawamorita, N, Tyagi, V., Sugino, S., Chancellor, M.B., Yoshimura, N., Tyagi, P. (2013): Downregulation of NGF expression in the bladder by antisense oligoucleotides as new treatment for overactive bladder. J urol., 190: 757-764. PMC3734554

15. Kashyap, M., Pore S, Yoshimura, N., Tyagi, P (2016). Constitutive expression Of NGF And P75(NTR) affected by bladder distension and NGF antisense treatment. Life Sciences. 148:93-8. PMC4792733.

16. Kashyap, M., Pore S, de Groat WC, Chermansky CJ, Yoshimura, N. Tyagi, P (2018). BDNF Overexpression in Bladder Induces Neuronal Changes To Mediate Bladder Overactivity. Am J Physiol Renal Physiol. 315(1):F45-F56.PMC6087795

Detrusor Underactivity/Underactive Bladder:  My lab characterized the functional and molecular changes leading to bladder overactivity following the exogenous overexpression of miR-132 in young rat (work won best poster at the AUA-2017 meeting). These biological effects of miR-132 in young rat could have potential therapeutic relevance in improving bladder emptying. Previously, I lead a diverse team towards basic research on underactive bladder, which linked the impairment of afferent signaling to the lower expression of neurotrophins and prostaglandins in rat model of UAB secondary to neuronal injury or diabetes. We identified the expression of miR-221 and miR-125b in OAB patients as key molecular determinants for the risk of urinary retention following Onabotulinum toxin injection and assessed the excitatory effect of Actotiamide in bladder as a potential treatment for underactive bladder. This documented experience in molecular research across the translational continuum from laboratory benchtop to bedside will be valuable to the current proposal.

17. Kashyap, M., Pore S, Chancellor MB, Yoshimura, N., Tyagi, P (2016). Bladder Overactivity Involves Over expression of MicroRNA 132 and Nerve Growth Factor. Life Sciences. 2016; 167:98- 104. PMC5125912. work on this project won best poster award at the AUA

18. Chermansky CJ, Kadow B, Kashyap, M., Tyagi, P (2017). MicroRNAs as Potential Biomarkers to Predict the Risk of Urinary Retention Following Intradetrusor Onabotulinumtoxin-A Injection. Neurourology Urodynamics. 37(1):99-105. PMID: 28493606

19. Singh N, Mizoguchi S, Suzuki T, Zabbarova I, Ikeda Y, Kanai A, Chermansky CJ, Yoshimura N, Tyagi P (2020). Excitatory effect of acotiamide on rat and human bladder: Implications for underactive bladder treatment. Life Sci; 258:118179. PMID:32758626

20. Kashyap, M., Singh N, Chermansky CJ, Yoshimura, N, Tyagi, P (2020) Constitutively active HCN channels constrain detrusor excitability and modulate evoked contractions of human bladder. Am J Clin Exp Urol. 8(5) (in press)

To summarize – proposed research on MRI and molecular targeted therapy will advance the understanding of BPH/LUTS

See Complete List of Publications