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1. Rationale and design of the Kidney Precision Medicine Project

Author

Ian H. de Boer, Charles E. Alpers, Evren U. Azeloglu, Ulysses G.J. Balis, Jonathan M. Barasch, Laura Barisoni, Kristina N. Blank, Andrew S. Bomback, Keith Brown, Pierre C. Dagher, Ashveena L. Dighe, Michael T. Eadon, Tarek M. El-Achkar, Joseph P. Gaut, Nir Hacohen, Yongqun He, Jeffrey B. Hodgin, Sanjay Jain, John A. Kellum, Krzysztof Kiryluk, Richard Knight, Zoltan G. Laszik, Chrysta Lienczewski, Laura H. Mariani, Robyn L. McClelland, Steven Menez, Dennis G. Moledina, Sean D. Mooney, John F. O’Toole, Paul M. Palevsky, Chirag R. Parikh, Emilio D. Poggio, Sylvia E. Rosas, Matthew R. Rosengart, Minnie M. Sarwal, Jennifer A. Schaub, John R. Sedor, Kumar Sharma, Becky Steck, Robert D. Toto, Olga G. Troyanskaya, Katherine R. Tuttle, Miguel A. Vazquez, Sushrut S. Waikar, Kayleen Williams, Francis Perry Wilson, Kun Zhang, Ravi Iyengar, Matthias Kretzler, Jonathan Himmelfarb, Stewart Lecker, Isaac Stillman, Sushrut Waikar, Gearoid Mcmahon, Astrid Weins, Samuel Short, Paul Hoover, Mark Aulisio, Leslie Cooperman, Leal Herlitz, John O’Toole, Emilio Poggio, John Sedor, Stacey Jolly, Paul Appelbaum, Olivia Balderes, Jonathan Barasch, Andrew Bomback, Pietro A. Canetta, Vivette D. d’Agati, Satoru Kudose, Karla Mehl, Jai Radhakrishnan, Chenhua Weng, Theodore Alexandrov, Tarek Ashkar, Daria Barwinska, Pierre Dagher, Kenneth Dunn, Michael Eadon, Michael Ferkowicz, Katherine Kelly, Timothy Sutton, Seth Winfree, Chirag Parikh, Avi Rosenberg, Pam Villalobos, Rubab Malik, Derek Fine, Mohammed Atta, Jose Manuel Monroy Trujillo, Alison Slack, Sylvia Rosas, Mark Williams, Evren Azeloglu, Cijang (John) He, Jens Hansen, Samir Parikh, Brad Rovin, Chris Anderton, Ljiljana Pasa-Tolic, Dusan Velickovic, Jessica Lukowski, George (Holt) Oliver, Joseph Ardayfio, Jack Bebiak, Taneisha Campbell, Catherine Campbell, Lynda Hayashi, Nichole Jefferson, Robert Koewler, Glenda Roberts, John Saul, Anna Shpigel, Edith Christine Stutzke, Lorenda Wright, Leslie Miegs, Roy Pinkeney, Rachel Sealfon, Olga Troyanskaya, Katherine Tuttle, Dejan Dobi, Yury Goltsev, Blue Lake, Maria Joanes, Zoltan Laszik, Andrew Schroeder, Minnie Sarwal, Tara Sigdel, Ulysses Balis, Victoria Blanc, Oliver He, Jeffrey Hodgin, Laura Mariani, Rajasree Menon, Edgar Otto, Jennifer Schaub, Sean Eddy, Michele Elder, Daniel Hall, John Kellum, Mary Kruth, Raghav Murugan, Paul Palevsky, Parmjeet Randhawa, Matthew Rosengart, Sunny Sims-Lucas, Mary Stefanick, Stacy Stull, Mitchell Tublin, Charles Alpers, Ian de Boer, Ashveena Dighe, Robyn Mcclelland, Sean Mooney, Stuart Shankland, Kristina Blank, Jonas Carson, Frederick Dowd, Zach Drager, Christopher Park, Guanshi Zhang, Shweta Bansal, Manjeri Venkatachalam, Asra Kermani, Simon Lee, Christopher Lu, Tyler Miller, Orson Moe, Harold Park, Kamalanathan Sambandam, Francisco Sanchez, Jose Torrealba, Toto Robert, Miguel Vazquez, Nancy Wang, Joe Gaut, Anitha Vijayan, Randy Luciano, Dennis Moledina, Ugwuowo Ugochukwu, and Sandy Alfano

Subjects
0301 basic medicine, Adult, Proteomics, medicine.medical_specialty, 030232 urology & nephrology, Subgroup analysis, Disease, urologic and male genital diseases, Kidney, Article, 03 medical and health sciences, 0302 clinical medicine, Diabetes mellitus, medicine, Humans, Prospective Studies, Precision Medicine, Renal Insufficiency, Chronic, Intensive care medicine, Prospective cohort study, business.industry, urogenital system, Acute kidney injury, Acute Kidney Injury, Precision medicine, medicine.disease, female genital diseases and pregnancy complications, 030104 developmental biology, medicine.anatomical_structure, Nephrology, business, Kidney disease
Abstract

Chronic kidney disease (CKD) and acute kidney injury (AKI) are common, heterogeneous, and morbid diseases. Mechanistic characterization of CKD and AKI in patients may facilitate a precision medicine approach to prevention, diagnosis, and treatment. The Kidney Precision Medicine Project aims to ethically and safely obtain kidney biopsies from participants with CKD or AKI, create a reference kidney atlas, and characterize disease subgroups to stratify patients based on molecular features of disease, clinical characteristics, and associated outcomes. An additional aim is to identify critical cells, pathways, and targets for novel therapies and preventive strategies. This project is a multicenter prospective cohort study of adults with CKD or AKI who undergo a protocol kidney biopsy for research purposes. This investigation focuses on kidney diseases that are most prevalent and therefore substantially burden the public health, including CKD attributed to diabetes or hypertension and AKI attributed to ischemic and toxic injuries. Reference kidney tissues (for example, living kidney donor biopsies) will also be evaluated. Traditional and digital pathology will be combined with transcriptomic, proteomic, and metabolomics analysis of the kidney tissue as well as deep clinical phenotyping for supervised and unsupervised subgroup analysis and systems biology analysis. Participants will be followed prospectively for ten years to ascertain clinical outcomes. Cell types, locations, and functions will be characterized in health and disease in an open, searchable, online kidney tissue atlas. All data from the Kidney Precision Medicine Project will be made readily available for broad use by scientists, clinicians, and patients.

Published
2021

2. Large-scale, three-dimensional tissue cytometry of the human kidney: a complete and accessible pipeline

Author

Michael J. Ferkowicz, Seth Winfree, Angela R. Sabo, Malgorzata M. Kamocka, Suraj Khochare, Daria Barwinska, Michael T. Eadon, Ying-Hua Cheng, Carrie L. Phillips, Timothy A. Sutton, Katherine J. Kelly, Pierre C. Dagher, Tarek M. El-Achkar, Kenneth W. Dunn, Richard Knight, Stewart Lecker, Isaac Stillman, Gearoid Mcmahon, Sus Waikar, Astrid Weins, Nir Hacohen, Paul Hoover, Mark Aulisio, Leslie Cooperman, Leal Herlitz, John O'toole, Emilio Poggio, John Sedor, Paul Appelbaum, Jonathan Barasch, Andrew Bomback, Vivette D'agati, Krzysztof Kiryluk, Karla Mehl, Ning (Sunny) Shang, Chenhua Weng, Laura Barisoni, Theodore Alexandrov, Tarek Ashkar, Pierre Dagher, Kenneth Dunn, Michael Eadon, Michael Ferkowicz, Katherine Kelly, Timothy Sutton, Steven Menez, Chirag Parikh, Avi Rosenberg, Pam Villalobos, Alison Slack, Sylvia Rosas, Mark Williams, Evren Azeloglu, Cijang (John) He, Ravi Iyengar, Samir Parikh, Chris Anderton, Ljiljana Pasa-Tolic, Dusan Velickovic, George (Holt) Oliver, Joseph Ardayfio, Jack Bebiak, Keith Brown, Taneisha Campbell, Catherine Campbell, Lynda Hayashi, Nichole Jefferson, Robert Koewler, Glenda Roberts, John Saul, Anna Shpigel, Edith Christine Stutzke, Lorenda Wright, Leslie Miegs, Roy Pinkeney, Rachel Sealfon, Olga Troyanskaya, Katherine Tuttle, Yury Goltsev, Blue Lake, Kun Zhang, Dejan Dobi, Maria Joanes, Zoltan Laszik, Garry Nolan, Andrew Schroeder, Ulysses Balis, Oliver He, Jeffrey Hodgin, Matthias Kretzler, Laura Mariani, Rajasree Menon, Edgar Otto, Jennifer Schaub, Becky Steck, Michele Elder, Daniel Hall, John Kellum, Mary Kruth, Raghav Murugan, Paul Palevsky, Parmjeet Randhawa, Matthew Rosengart, Sunny Sims-Lucas, Mary Stefanick, Stacy Stull, Mitchell Tublin, Charles Alpers, Ian De Boer, Malia Fullerton, Jonathan Himmelfarb, Robyn Mcclelland, Sean Mooney, Stuart Shankland, Kayleen Williams, Kristina Blank, Ashveena Dighe, Jonas Carson, Frederick Dowd, Zach Drager, Kumar Sharma, Guanshi Zhang, Asra Kermani, Simon Lee, Christopher Lu, Tyler Miller, Orson Moe, Harold Park, Kamalanathan Sambandam, Francisco Sanchez, Jose Torrealba, Toto Robert, Miguel Vazquez, Nancy Wang, Joe Gaut, Sanjay Jain, Anitha Vijayan, Randy Luciano, Dennis Moledina, Ugwuowo Ugochukwu, and Francis Perry Wilson

Subjects
0301 basic medicine, Computer science, Confocal, Cytological Techniques, Context (language use), Kidney, Article, Pathology and Forensic Medicine, law.invention, Flow cytometry, 03 medical and health sciences, 0302 clinical medicine, Imaging, Three-Dimensional, Confocal microscopy, law, Microscopy, medicine, Humans, Image analysis, Molecular Biology, Fluorescent Dyes, Microscopy, Confocal, medicine.diagnostic_test, Cell Biology, Autofluorescence, 030104 developmental biology, Microscopy, Fluorescence, Multiphoton, 030220 oncology & carcinogenesis, Cytometry, Software, Biomedical engineering
Abstract

The advent of personalized medicine has driven the development of novel approaches for obtaining detailed cellular and molecular information from clinical tissue samples. Tissue cytometry is a promising new technique that can be used to enumerate and characterize each cell in a tissue and, unlike flow cytometry and other single-cell techniques, does so in the context of the intact tissue, preserving spatial information that is frequently crucial to understanding a cell’s physiology, function, and behavior. However, the wide-scale adoption of tissue cytometry as a research tool has been limited by the fact that published examples utilize specialized techniques that are beyond the capabilities of most laboratories. Here we describe a complete and accessible pipeline, including methods of sample preparation, microscopy, image analysis, and data analysis for large-scale three-dimensional tissue cytometry of human kidney tissues. In this workflow, multiphoton microscopy of unlabeled tissue is first conducted to collect autofluorescence and second-harmonic images. The tissue is then labeled with eight fluorescent probes, and imaged using spectral confocal microscopy. The raw 16-channel images are spectrally deconvolved into 8-channel images, and analyzed using the Volumetric Tissue Exploration and Analysis (VTEA) software developed by our group. We applied this workflow to analyze millimeter-scale tissue samples obtained from human nephrectomies and from renal biopsies from individuals diagnosed with diabetic nephropathy, generating a quantitative census of tens of thousands of cells in each. Such analyses can provide useful insights that can be linked to the biology or pathology of kidney disease. The approach utilizes common laboratory techniques, is compatible with most commercially-available confocal microscope systems and all image and data analysis is conducted using the VTEA image analysis software, which is available as a plug-in for ImageJ. Tissue cytometry is a promising new microscopy technique that can be used to enumerate and characterize each cell in a tissue. Here the authors describe a complete and accessible pipeline, including methods of sample preparation, microscopy, image analysis, and data analysis for large-scale three-dimensional tissue cytometry of human kidney tissues.

Published
2020

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