Circulating uromodulin inhibits systemic oxidative stress by inactivating the TRPM2 channel

LaFavers, Kaice A.; Macedo, Etienne; Garimella, Pranav S.; Lima, Camila; Khan, Shehnaz; Myslinski, Jered; McClintick, Jeanette N.; Witzmann, Frank A.; Winfree, Seth; Phillips, Carrie L.; Hato, Takashi; Dagher, Pierre C.; Wu, Xue-Ru; El‐Achkar, Tarek M.; Micanovic, Radmila

doi:10.1126/scitranslmed.aaw3639

Cited by 67 publications

(63 citation statements)

References 73 publications

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“…These techniques, such as transcriptomics, proteomics, and large-scale quantitative 3-dimensional (3D) imaging, can provide an unbiased and comprehensive approach to discovery. [6][7][8][9][10] We present here a detailed study contrasting CaOx and brushite clinical stone phenotypes, using these orthogonal omics approaches.…”

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confidence: 99%

A Precision Medicine Approach Uncovers a Unique Signature of Neutrophils in Patients With Brushite Kidney Stones

Makki

Winfree

Lingeman

et al. 2020

Kidney International Reports

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Introduction: We have previously found that papillary histopathology differs greatly between calcium oxalate and brushite stone formers (SF); the latter have much more papillary mineral deposition, tubular cell injury, and tissue fibrosis. Methods: In this study, we applied unbiased orthogonal omics approaches on biopsied renal papillae and extracted stones from patients with brushite or calcium oxalate (CaOx) stones. Our goal was to discover stone type-specific molecular signatures to advance our understanding of the underlying pathogenesis. Results: Brushite SF did not differ from CaOx SF with respect to metabolic risk factors for stones but did exhibit increased tubule plugging in their papillae. Brushite SF had upregulation of inflammatory pathways in papillary tissue and increased neutrophil markers in stone matrix compared with those with CaOx stones. Large-scale 3-dimensional tissue cytometry on renal papillary biopsies showed an increase in the number and density of neutrophils in the papillae of patients with brushite versus CaOx, thereby linking the observed inflammatory signatures to the neutrophils in the tissue. To explain how neutrophil proteins appear in the stone matrix, we measured neutrophil extracellular trap (NET) formation-NETosis-and found it significantly increased in the papillae of patients with brushite stones compared with CaOx stones. Conclusion: We show that increased neutrophil infiltration and NETosis is an unrecognized factor that differentiates brushite and CaOx SF and may explain the markedly increased scarring and inflammation seen in the papillae of patients with brushite stones. Given the increasing prevalence of brushite stones, the role of neutrophil activation in brushite stone formation requires further study.

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confidence: 99%

A Precision Medicine Approach Uncovers a Unique Signature of Neutrophils in Patients With Brushite Kidney Stones

Makki

Winfree

Lingeman

et al. 2020

Kidney International Reports

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“…However, it is currently unknown how pUMOD activates endothelial cells and innate immune cells, thereby mounting an immune response. Several UMOD receptors have been described including TLR4, Transient receptor potential melastatin 2 (TRPM2) and TRPM6 channels (13,16,31). TRPM6 is neither expressed on endothelial cells nor on neutrophils, suggesting TLR4…”

Section: Neonatal Obstructive Nephropathy Induces Intra-and Extratubumentioning

confidence: 99%

“…Reduced serum levels correlate with higher cardiac mortality (15), elevated systemic ROS (16) and increased susceptibility to develop type 2 diabetes mellitus accompanied by reduced glucose metabolism (17). In addition, low serum levels were shown to be associated with reduced GFR (18), linking mUMOD serum levels to kidney function and survival.…”

Section: Introductionmentioning

confidence: 99%

Extratubular polymerized uromodulin induces leukocyte recruitment and inflammationin vivo

Immler¹,

Lange-Sperandio

Steffen³

et al. 2020

Preprint

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Uromodulin (UMOD) is produced and secreted by tubular epithelial cells. Secreted UMOD polymerizes (pUMOD) within the lumen, where it regulates salt transport and protects the kidney from bacteria and stone formation. Under various pathological conditions, pUMOD accumulates within the tubular lumen and reaches extratubular sites where it may interact with renal interstitial cells. Here, we investigated the potential of extratubular pUMOD to act as a damage associated molecular pattern (DAMP) molecule thereby creating local inflammation. We found that intrascrotal and intraperitoneal injection of pUMOD induced leukocyte recruitment in vivo and led to TNF-α secretion by F4/80 positive macrophages. Additionally, pUMOD directly affected vascular permeability and increased neutrophil extravasation independent of macrophage-released TNF-α. Interestingly, pUMOD did not directly upregulate adhesion molecules on endothelial cells and did not directly activate β2 integrins on neutrophils. In obstructed neonatal murine kidneys, we observed extratubular UMOD accumulation with tubular atrophy and leukocyte infiltrates. Finally, we found extratubular UMOD deposits associated with peritubular leukocyte infiltration in kidneys from patients with inflammatory kidney diseases. Taken together, we identified extratubular pUMOD as a strong inducer of leukocyte recruitment, underlining its critical role in mounting an inflammatory response in various kidneys pathologies.

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“…Each of these technologies uses different approaches in sample preparation, dissociation or processing that have unique advantages and disadvantages (see TIS protocol on https://kpmp.org/researcher-resources/). Given the potential to introduce artifacts or miss cell types due to processing of these specimens, concurrent bulk RNA expression analysis on undissociated tissue or regional laser microdissection (LMD) (IU/OSU) is performed (16)(17)(18). Using the LMD technology, transcriptomic signatures are identified for seven sub-segments of the nephron including the proximal tubule (PT), thick ascending loop of Henle (TAL), distal convoluted tubule (DCT), collecting duct (CD), as well as compartment-specific signatures for glomeruli, the tubulo-interstitium (TI), the interstitium (without glomeruli or tubules), as well as a bulk cross-section of the entire biopsy.…”

Section: Overview Of Kpmp Technologies: Creating a Molecular And Funcmentioning

confidence: 99%

A Multimodal and Integrated Approach to Interrogate Human Kidney Biopsies with Rigor and Reproducibility: The Kidney Precision Medicine Project

El-Achkar

Eadon

Menon

et al. 2019

Preprint

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Comprehensive and spatially mapped molecular atlases of organs at a cellular level are a critical resource to gain insights into pathogenic mechanisms and therapies tailored to the disease of each patient. Obtaining rigorous and reproducible results from disparate methods and at different sites to interrogate biomolecules at a single cell level or in 3-dimensional (3D) space is a significant challenge that can be a futile exercise if not well controlled. The Kidney Precision Medicine Project (KPMP) is an endeavor to generate 3D molecular atlases of healthy and diseased kidney biopsies using multiple state-of-the-art OMICS and imaging technologies across several institutions. We describe a pipeline for generating a reliable and authentic single cell/region 3D molecular atlas of human adult kidney with emphasis on quality assurance, quality control, validation and harmonization across different OMICS and imaging methods. Our “follow the tissue” approach encompasses sample procurement to data generation, analysis, data sharing, while standardizing, and harmonizing procedures at sample collection, processing, storage and shipping. We provide key features of preanalytical parameters, bioassays, post analyses, reference standards and data depositions. A pilot experiment from a common source tissue processed and analyzed at different institutions was executed to identify potential sources of variation, feasibility of multimodal analyses and unique and redundant features of the macromolecules being characterized by each technology. An important outcome was identification of limitations and strengths of the different technologies, and how composite information can be leveraged for clinical application. A peer review system was established to critically review quality control measures and the reproducibility of data generated by each technology before granting approval to work on clinical biopsy specimens. This unique pipeline establishes a process that economizes the use of valuable biopsy tissue for multi-OMICS and imaging analysis with stringent quality control to ensure rigor and reproducibility of results and which can serve as a model for similar personalized medicine projects.Author ContributionsMTE, RM, BBL, TS, TA, AS, SP, CRA, DD, EAO, SW, GZ, MJ and KD performed the ground work for the quality control group, wrote the KPMP TIS manual of procedures and generated figures. HH, JZ, RS, RM, TS, EAO, MTE, TME, PH, SP, MK, ZL and SJ generated the initial working reference marker list. CEA, TME, JBH, JL, MK and SJ led the Pilot 1 protocol. TME, VD, LB, JG, CEA, ZL, SJ and JBH developed the pathology QC tissue qualification and tissue processing criteria. JBH organized and executed the Pilot tissue collection and distribution. JC designed the SpecTrack system. CP prepared and organized the TIS manual of procedures and performed data organization services. YH led ontology development for QC metadata and knowledge standardization. BS and EA organized data integration efforts and data authentication in the data hub. KS and MS led the OMICS discussion group. SJ led the quality control group. TME and CEA led the tissue processing group. MTE and SJ led the Molecular and Pathology Integration group. MTE and SJ conceived and led the TISAC process. RI, OGT KZ, ZL, PH, BR, PCD, KS, MS, JBH, CEA, LB, JG, TME, MK and SJ conceived the integrated TIS pipeline and QC vision. TME and SJ wrote the initial draft of the paper. All authors contributed to the writing and editing of the manuscript.

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Circulating uromodulin inhibits systemic oxidative stress by inactivating the TRPM2 channel

Cited by 67 publications

References 73 publications

A Precision Medicine Approach Uncovers a Unique Signature of Neutrophils in Patients With Brushite Kidney Stones

A Precision Medicine Approach Uncovers a Unique Signature of Neutrophils in Patients With Brushite Kidney Stones

Extratubular polymerized uromodulin induces leukocyte recruitment and inflammationin vivo

A Multimodal and Integrated Approach to Interrogate Human Kidney Biopsies with Rigor and Reproducibility: The Kidney Precision Medicine Project

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