Design and Methods of the Validating Injury to the Renal Transplant Using Urinary Signatures (VIRTUUS) Study in Children

Juhi Kumar¹, Kévin Contrepois², Michael Snyder², Paul C Grimm³, Asha Moudgil⁴, Jodi M Smith⁵, Amy E Bobrowski⁶, Priya S Verghese⁷, David Hooper⁸, Elizabeth Ingulli⁹, Rachel Lestz¹⁰, Patricia Weng¹¹, Janaiya L Reason¹², Tom D Blydt-Hansen¹³, Manikkam Suthanthiran¹⁴, Brendan Keating¹⁵, Sandra Amaral¹⁶

Affiliations

¹ Department of Pediatrics and Population Health Sciences, Weill Cornell Medicine, New York, NY.
² Department of Genetics, Stanford University School of Medicine, Stanford, CA.
³ Department of Pediatrics, Stanford University School of Medicine, and Lucile Packard Children's Hospital, Stanford, CA.
⁴ Division of Nephrology, Children's National Hospital, The George Washington University School of Medicine and Health Sciences, Washington, DC.
⁵ Seattle Children's Hospital, University of Washington, Seattle, WA.
⁶ Center for Pediatric Nephrology, Cleveland Clinic Children's, Cleveland, OH.
⁷ Ann and Robert H. Lurie Children's Hospital, Chicago, IL.
⁸ Cincinnati Children's Hospital, Cincinnati, OH.
⁹ Rady Children's, San Diego, CA.
¹⁰ Children's Hospital Los Angeles, Los Angeles, CA.
¹¹ Mattel Children's Hospital, University of California Los Angeles, Los Angeles, CA.
¹² University of Pennsylvania, Philadelphia, PA.
¹³ University of British Columbia, Vancouver, BC, Canada.
¹⁴ Department of Medicine, Weill Cornell Medicine, New York, NY.
¹⁵ Division of Transplantation, Department of Surgery, Perelman School of Medicine, Penn Transplant Institute, University of Pennsylvania, Philadelphia, PA.
¹⁶ Departments of Pediatrics and Biostatistics, Epidemiology and Informatics, Perelman School of Medicine, Children's Hospital of Philadelphia and University of Pennsylvania, Philadelphia, PA.

PMID: 34805493
PMCID: PMC8601357
DOI: 10.1097/TXD.0000000000001244

Design and Methods of the Validating Injury to the Renal Transplant Using Urinary Signatures (VIRTUUS) Study in Children

Juhi Kumar et al. Transplant Direct. 2021.

. 2021 Nov 17;7(12):e791.

doi: 10.1097/TXD.0000000000001244. eCollection 2021 Dec.

Authors

Affiliations

¹ Department of Pediatrics and Population Health Sciences, Weill Cornell Medicine, New York, NY.
² Department of Genetics, Stanford University School of Medicine, Stanford, CA.
³ Department of Pediatrics, Stanford University School of Medicine, and Lucile Packard Children's Hospital, Stanford, CA.
⁴ Division of Nephrology, Children's National Hospital, The George Washington University School of Medicine and Health Sciences, Washington, DC.
⁵ Seattle Children's Hospital, University of Washington, Seattle, WA.
⁶ Center for Pediatric Nephrology, Cleveland Clinic Children's, Cleveland, OH.
⁷ Ann and Robert H. Lurie Children's Hospital, Chicago, IL.
⁸ Cincinnati Children's Hospital, Cincinnati, OH.
⁹ Rady Children's, San Diego, CA.
¹⁰ Children's Hospital Los Angeles, Los Angeles, CA.
¹¹ Mattel Children's Hospital, University of California Los Angeles, Los Angeles, CA.
¹² University of Pennsylvania, Philadelphia, PA.
¹³ University of British Columbia, Vancouver, BC, Canada.
¹⁴ Department of Medicine, Weill Cornell Medicine, New York, NY.
¹⁵ Division of Transplantation, Department of Surgery, Perelman School of Medicine, Penn Transplant Institute, University of Pennsylvania, Philadelphia, PA.
¹⁶ Departments of Pediatrics and Biostatistics, Epidemiology and Informatics, Perelman School of Medicine, Children's Hospital of Philadelphia and University of Pennsylvania, Philadelphia, PA.

PMID: 34805493
PMCID: PMC8601357
DOI: 10.1097/TXD.0000000000001244

Abstract

Lack of noninvasive diagnostic and prognostic biomarkers to reliably detect early allograft injury poses a major hindrance to long-term allograft survival in pediatric kidney transplant recipients.

Methods: Validating Injury to the Renal Transplant Using Urinary Signatures Children's Study, a North American multicenter prospective cohort study of pediatric kidney transplant recipients, aims to validate urinary cell mRNA and metabolite profiles that were diagnostic and prognostic of acute cellular rejection (ACR) and BK virus nephropathy (BKVN) in adult kidney transplant recipients in Clinical Trials in Organ Transplantation-4. Specifically, we are investigating: (1) whether a urinary cell mRNA 3-gene signature (18S-normalized CD3ε, CXCL10 mRNA, and 18S ribosomal RNA) discriminates biopsies with versus without ACR, (2) whether a combined metabolite profile with the 3-gene signature increases sensitivity and specificity of diagnosis and prognostication of ACR, and (3) whether BKV-VP1 mRNA levels in urinary cells are diagnostic of BKVN and prognostic for allograft failure.

Results: To date, 204 subjects are enrolled, with 1405 urine samples, including 144 biopsy-associated samples. Among 424 urine samples processed for mRNA, the median A260:280 ratio (RNA purity) was 1.91, comparable with Clinical Trials in Organ Transplantation-4 (median 1.82). The quality control failure rate was 10%. Preliminary results from urine supernatant showed that our metabolomics platform successfully captured a broad array of metabolites. Clustering of pool samples and overlay of samples from various batches demonstrated platform robustness. No study site effect was noted.

Conclusions: Multicenter efforts to ascertain urinary biomarkers in pediatric kidney transplant recipients are feasible with high-quality control. Further study will inform whether these signatures are discriminatory and predictive for rejection and infection.

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Conflict of interest statement

M.Snyder is a cofounder and on the advisory board of Personalis, SensOmics, January, Filtricine, Qbio, Protos, and Mirive. M.Snyder is also on the advisory board of Genapsys and Tailai. J.K. is a one-time Pediatric Advisory Board attendee for Reata Pharmaceuticals, Inc. The other authors declare no conflicts of interest.

Figures

**FIGURE 1.**
VIRTUUS Clinical Sites and Core Labs across the United States and Canada. VIRTUUS, Validating Injury to the Renal Transplant Using Urinary Signatures.

See this image and copyright information in PMC

References

1. Hart A, Smith JM, Skeans MA, et al. . OPTN/SRTR 2018 annual data report: kidney. Am J Transplant. 2020;20(suppl s1):20–130. - PubMed
1. Smith JM, Ho PL, McDonald RA; North American Pediatric Renal Transplant Cooperative Study. Renal transplant outcomes in adolescents: a report of the North American Pediatric Renal Transplant Cooperative Study. Pediatr Transplant. 2002;6:493–499. - PubMed
1. Blydt-Hansen TD, Sharma A, Gibson IW, et al. . Validity and utility of urinary CXCL10/Cr immune monitoring in pediatric kidney transplant recipients. Am J Transplant. 2021;21:1545–1555. - PubMed
1. Blydt-Hansen TD, Sharma A, Gibson IW, et al. . Urinary metabolomics for noninvasive detection of antibody-mediated rejection in children after kidney transplantation. Transplantation. 2017;101:2553–2561. - PubMed
1. Fan S, Kind T, Cajka T, et al. . Systematic error removal using random forest for normalizing large-scale untargeted lipidomics data. Anal Chem. 2019;91:3590–3596. - PMC - PubMed

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Design and Methods of the Validating Injury to the Renal Transplant Using Urinary Signatures (VIRTUUS) Study in Children

Affiliations

Design and Methods of the Validating Injury to the Renal Transplant Using Urinary Signatures (VIRTUUS) Study in Children

Authors

Affiliations

Abstract

Conflict of interest statement

Figures

References

Grants and funding

LinkOut - more resources

Full Text Sources