IDENTIFICATION OF BLOOD BIOMARKER CANDIDATES IN CALCIPHYLAXIS PATIENTS RECEIVING STEM CELL THERAPY USING PROTEOMICS AND A HUMAN MICROVASCULAR CHIP PLATFORM

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https://storage.unitedwebnetwork.com/files/1099/39c018ad1c986b077a425365df02fe11.pdf

Abstract Title *

IDENTIFICATION OF BLOOD BIOMARKER CANDIDATES IN CALCIPHYLAXIS PATIENTS RECEIVING STEM CELL THERAPY USING PROTEOMICS AND A HUMAN MICROVASCULAR CHIP PLATFORM

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Co-author 1

Ningning Wang wangnn@njmu.edu.cn the First Affiliated Hospital with Nanjing Medical University, Jiangsu Province Hospital Division of Nephrology, Department of Geriatrics Nanjing China *

Co-author 2

Jiaying Hu hujiaying0727@163.com the First Affiliated Hospital with Nanjing Medical University, Jiangsu Province Hospital Division of Nephrology, Department of Geriatrics Nanjing China -

Co-author 3

Shijiu Lu lsj10743@163.com the First Affiliated Hospital with Nanjing Medical University, Jiangsu Province Hospital Division of Nephrology, Department of Geriatrics Nanjing China -

Co-author 4

Lianju Qin ljqin@njmu.edu.cn the First Affiliated Hospital with Nanjing Medical University, Jiangsu Province Hospital State Key Laboratory of Reproductive Medicine and Offspring Health, Center of Clinical Reproductive Medicine Nanjing China -

Co-author 5

Yaoting Sun sunyaoting@westlake.edu.cn Westlake University School of Medicine Hangzhou China -

Co-author 6

Xiaoxue Ye 13057667895@163.com the First Affiliated Hospital with Nanjing Medical University, Jiangsu Province Hospital Division of Nephrology, Department of Geriatrics Nanjing China -

Co-author 7

Qinyi Lin 1030921469@qq.com the First Affiliated Hospital with Nanjing Medical University, Jiangsu Province Hospital Division of Nephrology, Department of Geriatrics Nanjing China -

Co-author 8

Jiayin Liu jyliu_nj@126.com the First Affiliated Hospital with Nanjing Medical University, Jiangsu Province Hospital State Key Laboratory of Reproductive Medicine and Offspring Health, Center of Clinical Reproductive Medicine, Nanjing China -

Co-author 9

Yun Liu liuyun@njmu.edu.cn the First Affiliated Hospital with Nanjing Medical University, Jiangsu Province Hospital Division of Endocrinology, Department of Geriatrics, Nanjing China -

Co-author 10

Shihui Xu xush@avatarget.com.cn Jiangsu Avatarget Biotechnology Co., Ltd. Jiangsu Avatarget Biotechnology Co., Ltd Nanjing China -

Co-author 11

Zaozao Chen czz@avatarget.com.cn Southeast University State Key Laboratory of Digital Medical Engineering, School of Biological Science and Medical Engineering Nanjing China -

Co-author 12

Tiannan Guo guotiannan@westlake.edu.cn Westlake University School of Medicine Hangzhou China -

Co-author 13

Yi Zhu zhuyi@westlake.edu.cn Westlake University School of Medicine Hangzhou China -

Co-author 14

Co-author 15

Introduction

Calciphylaxis, also known as calcific uremic arteriolopathy (CUA), is a rare and high fatality rate disease with skin ischemia and necrosis. It is imperative to identify novel biomarkers for treatment guidance.

Methods

We rescued CUA patients with human amnion-derived mesenchymal stem cells (hAMSCs). In a discovery cohort including uremic patients (n=10) and CUA patients (n=3), plasma proteomic analysis was investigated for the differentially expressed proteins (DEPs). Plasma core DEPs were measured by ELISA. Skin tissue was analyzed for calcification and target proteins by multiplex immunofluorescence. Due to the lack of animal models related to calciphylaxis, we developed a microvascular chip to evaluate the damage caused by biomarker and the protective effects of hAMSCs.

Results

Plasma proteomics (discovery cohort: 3 CUA, 10 uremic) revealed decreased Thrombospondin 1(THBS1) and Latent TGF-β binding protein 1(LTBP1) after hAMSC therapy, linked to coagulation and wound healing. In vitro, blocking THBS1/TGF-β1 impaired endothelial adhesion and coagulation. ELISA in a validation cohort (8 CUA, 20 uremic) confirmed elevated THBS1/TGF-β1 in calciphylaxis, reduced post-treatment (6 patients), but rebounded with infrequent therapy (2 patients). Multiplex immunofluorescence showed THBS1 and CD47 co-localized with CD31 and integrin β3(ITGB3) in injuried microvessels. The human microvascular chip demonstrated that THBS1 inhibition or hAMSC-conditioned medium alleviates microvascular injury under uremic conditions（Fig 1).

Conclusion

These findings suggest THBS1/TGF-β1 as potential biomarkers for calciphylaxis and support hAMSC therapy as a promising regenerative approach（Fig 2).

The content presented in this abstract was discussed at the International Symposium on Microphysiological Systems (MPS 2025).

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