CPTCs Drive Somatic-Visceral Communication via the Wnt Axis in Somatic Mechanotherapy: A Single-Cell Deep Learning Study

Somatic mechanical stimulation (e.g., acupuncture) exerts systemic immunomodulatory effects, yet the cellular bridge translating peripheral physical force into visceral repair remains elusive. Here, employing a custom interpretable deep learning framework (CARSS) on single-cell RNA sequencing data, we identify CD34$^{+}$PDGFR$α$$^{+}$ telocytes (CPTCs) as the primary mechanosensors in both fascia and colon during bacterial colitis. We show that somatic mechanotherapy triggers an AP-1/Hsp70-dependent transcriptional program in fascial CPTCs, inducing systemic Wnt elevation, which elicits a “transcriptional resonance” in colonic CPTCs, reprogramming their communication network from an inflammatory amplifier to a Wnt-driven regenerative hub. Mechanistically, this axis activates epithelial $β$-catenin/Myc signaling, suppressing apoptosis and restoring barrier integrity independent of immune cells. Our findings define a CPTC-Driven Mechano-Resonance Axis, where CPTCs serve as synchronized relay stations that convert local mechanical cues into systemic regenerative microenvironments.

💡 Research Summary

This study investigates how localized mechanical stimulation, such as acupuncture, can produce systemic regenerative effects, focusing on the cellular conduit that translates peripheral force into distal organ repair. The authors combined a novel interpretable deep‑learning pipeline—Cellular Acupuncture Response Scoring System (CARSS)—with high‑resolution single‑cell RNA sequencing of both subcutaneous fascia and colon from a rat model of Salmonella‑induced colitis.

In the animal model, eight‑week‑old female Sprague‑Dawley rats were infected orally with Salmonella typhimurium and then randomized to receive daily 20‑minute acupuncture at three abdominal midline points (CV4, CV6, CV12) for seven days, or to a restraint‑only control. After treatment, fascia and colon tissues were harvested for histology, electron microscopy, ELISA, and single‑cell transcriptomics.

Single‑cell analysis identified 15 major cell types in each tissue. Among them, a CD34⁺PDGFRα⁺ telocyte population—named CPTC (mechanosensitive telocyte)—was present in both fascia and colon. CPTCs displayed long, thin processes and expressed the mechanosensitive channel Piezo2.

CARSS reframes the problem of “which cells respond to acupuncture” as a machine‑learning generalization task. Adaptive multilayer perceptron (MLP) models with 4–8 hidden layers, 20–128 neurons per layer, and various activation functions (ReLU, ELU, Hardswish) were trained on a 90:10 train‑test split. A two‑stage grid search explored 22,680 hyper‑parameter combinations. Model performance was evaluated in two ways: (1) “Self‑matrix” testing on pure CPTC data, and (2) “Generalization” testing on mixed‑cell datasets containing all identified cell types. The ratio of generalization accuracy to self‑matrix accuracy (G‑Index) quantified how robustly a cell type’s response signature persisted amid cellular heterogeneity. CPTCs achieved the highest G‑Index, indicating they are the primary mechanosensitive population.

Interpretability was achieved with DeepSHAP. The top 100 SHAP‑ranked genes were enriched for AP‑1 transcription factors (Fos, Jun), heat‑shock protein Hsp70 (Hspa1a/b), the mechanosensor Piezo2, and the secreted ligand Wnt2. Gene‑set enrichment analysis highlighted pathways related to mechanical stimulus response, extracellular vesicle signaling, and canonical Wnt signaling.

Functional validation showed that acupuncture markedly increased serum Wnt2 levels (≈2.3‑fold). In the colon, Wnt2 elevation triggered nuclear translocation of β‑catenin, up‑regulation of Myc, and a reduction in TUNEL‑positive apoptotic cells. Importantly, depletion of CD45⁺ immune cells did not abolish these effects, indicating that the regenerative cascade operates largely independent of immune mediation.

In vitro, CPTCs cultured from fascia were co‑cultured with human colonic epithelial cells (NCM460) in a transwell system. When Salmonella was added (MOI = 1) to the epithelial compartment, CPTC‑derived extracellular vesicles containing Wnt2 suppressed bacterial colony formation and restored tight‑junction proteins (ZO‑1, Occludin). Pharmacological blockade of Wnt secretion (IWP‑2) or β‑catenin activity (XAV‑939) prevented Myc induction and barrier recovery, confirming the necessity of the Wnt/β‑catenin/Myc axis.

Cross‑tissue homology analysis using Harmony‑corrected integration demonstrated that fascial and colonic CPTCs share a highly overlapping transcriptomic signature, supporting the concept of a “mechano‑resonance axis” in which mechanically activated fascia CPTCs broadcast a systemic Wnt signal that re‑programs distal CPTCs. CellCall and PySCENIC analyses revealed that, after acupuncture, fascia CPTCs shift from an inflammatory communication network (dominated by cytokine‑TNF signaling) to a regenerative network centered on Wnt ligand–receptor interactions.

Overall, the paper delivers three major contributions:

1. Methodological Innovation – CARSS provides a scalable, interpretable framework for pinpointing stimulus‑responsive cell populations within complex single‑cell datasets, overcoming the limitations of conventional differential‑expression pipelines.

2. Mechanistic Insight – The AP‑1/Hsp70‑driven secretion of Wnt2 from CD34⁺PDGFRα⁺ telocytes constitutes a non‑neural, hormone‑like conduit that translates localized mechanical cues into a systemic regenerative signal.

3. Therapeutic Implication – By demonstrating that Wnt/β‑catenin/Myc activation can restore epithelial integrity without immune cell involvement, the study suggests that targeted modulation of telocyte‑derived Wnt signaling could be harnessed for treating inflammatory bowel diseases or other organ injuries where mechanical therapy is feasible.

The authors make all raw sequencing data (GSA CRA020447, GEO GSE261489) and analysis scripts (GitHub) publicly available, facilitating reproducibility and future extensions of the CARSS platform to other mechanobiology contexts.