Calciphylaxis, also known as calcific uremic arteriolopathy (CUA), is an orphan disease without proven therapies, we rescued it with human amnion-derived mesenchymal stem cells (hAMSCs). In a discovery cohort of 10 uremic patients and 3 CUA patients, plasma proteomic analysis showed core differentially expressed proteins (DEPs) Thrombospondin 1 (THBS1) and Latent transforming growth factor (TGF)-β binding protein 1 (LTBP1) decreased significantly after 3 days of hAMSC treatment. Single-cell transcriptome sequencing of peripheral blood mononuclear cells (PBMCs) indicated megakaryocytes were the source of THBS1 in CUA patient. Same as the discovery cohort, plasma THBS1 and TGF-β1 levels were increased in seven CUA patients compared to the uremic group (n=20), as measured by enzyme-linked immunosorbent assay (ELISA) in the validation cohort. They can be inhibited after hAMSC treatment and increased as the frequency of therapy decreased. THBS1 and its receptor, CD47, were increased in the CUA skin. THBS1 and TGF-β1 are biomarker candidates for calciphylaxis.

The authors have declared no competing interest.

NCT04592640

This work was funded by the National Natural Science Foundation of China (81270408, 81570666, 81730041, and 81671447), National Key R&D Program of China (2022YF0608403, 2021YFA1301600), the International Society of Nephrology (ISN) Clinical Research Program (18-01-0247), CKD Anemia Research Foundation from China International Medical Foundation (Z-2017-24-2037), Natural Science Foundation of Jiangsu Province (BK20243054), Outstanding Young and Middle-Aged Talents Support Program of The First Affiliated Hospital of Nanjing Medical University (Jiangsu Province Hospital), Jiangsu Province Hospital (the First Affiliated Hospital with Nanjing Medical University) Clinical Capacity Enhancement Project (JSPH-MA-2023-7), the National Key Research and Development Program of China (2017YFC1001303), the State Key Laboratory of Reproductive Medicine Program (SKLRM-GC201803), the State Key Laboratory of Reproductive Medicine and Offspring Health Program (SKLRM-K202105), "Pioneer" and "Leading Goose" R&D Program of Zhejiang (2024SSYS0035) and Westlake Omics Junior Clinician Support Program 2021.

I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained.

Yes

The details of the IRB/oversight body that provided approval or exemption for the research described are given below:

This study was approved by the ethics committee of the First Affiliated Hospital of Nanjing Medical University in China (2018-QT-001, 2020-QT-01, 2020-QT-09, 2020-12-02).

I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals.

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I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance).

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I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable.

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The proteomic data has been deposited in iProX (IPX0009031001, https://www.iprox.cn/page/DSV021.html;?url=1718120623768Uh8P, password h84O) and will be made publicly available upon publication of this manuscript. The single-cell RNA sequencing data has been deposited in NGDC (OMIX006677, https://ngdc.cncb.ac.cn/omix/preview/iuik4gl4. No customized code in proteomics analysis. The single-cell transcriptome analysis code used in this study has been deposited at GitHub under https://github.com/lsj46/CUA-Code/blob/master/Code.Rmd. However, raw data for individual patients cannot be publicly disclosed due to patient confidentiality reasons. Researchers can apply for access to these data, subject to approval from the individual institutional review boards. Any additional information required to reanalyze the data reported in this work paper is available from the lead contact upon request.