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March 21, 2026Advanced Science0 citationsOpen Access

RENAL‐CHIP: Rejection Evaluation via Non‐Invasive Analysis of Circulating Podocytes With Herringbone‐Chip Isolation Platform

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JSJuan SongYLYing LiuZZZeyuan Zheng

Key Points

  • The study aims to evaluate the potential of circulating podocytes as biomarkers for acute renal allograft rejection.
  • Isolated donor-derived circulating podocytes using a herringbone microfluidic chip.
  • Analyzed blood samples from 65 participants, confirming rejection via biopsy.
  • Used fluorescence in situ hybridization (FISH) for donor origin verification.
  • Performed single-cell RNA sequencing on isolated circulating podocytes.
  • Transplant recipients with biopsy-confirmed rejection had ≥35 CPCs per mL.
  • Capture efficiency of the microfluidic chip was 84.4%; release was 95.5%; viability was 96.0%.
  • Single-cell RNA-seq revealed up-regulated immune response genes in rejecting versus stable samples.

Abstract

Immune rejection limits long-term renal allograft survival, yet current diagnostics lack non-invasive, and precise detection. Here, donor-derived circulating podocytes (CPCs) are identified as a blood-based surrogate of acute rejection. A magnetically reversible herringbone microfluidic chip equipped with epithelial cell adhesion molecule (EpCAM) antibody-functionalized magnetic beads efficiently isolates CPCs from 1 mL of peripheral blood (capture efficiency 84.4%, release 95.5%, viability 96.0%). Among 65 participants, only transplant recipients with biopsy-confirmed rejection exhibited≥35 CPCs mL- 1 (AUC = 1.0). Fluorescence in situ hybridization (FISH) of sex-mismatched transplants confirmed donor origin. Single-cell RNA-seq of 10 isolated CPCs revealed up-regulation of podocyte-injury and innate/adaptive immune genes (such as NF-κB, TNF, NOD-like receptor pathways) in rejection versus stable samples. CPC enumeration thus provides a minimally invasive, mechanistically informed warning of renal allograft rejection.

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Cite This Study

Song et al. (2026) studied this question.

synapsesocial.com/papers/69be35f96e48c4981c6747c8https://doi.org/10.1002/advs.202520426
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