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

The gist: CPTCs drive somatic–visceral communication via the Wnt axis in somatic mechanotherapy, where fascial CPTCs act as primary mechanosensors that translate mechanical cues into systemic regenerative signals in distal colonic CPTCs > <ref:2602.05451#pg14>

How it works

The study employs a custom interpretable deep learning framework named CARSS to identify CD34+PDGFRα+ telocytes (CPTCs) as the primary mechanosensors in both fascia and colon during bacterial colitis > <ref:2602.05451#pg18> The CARSS framework reframes the biological challenge of identifying mechanosensitive cells into a computational problem of evaluating machine learning model generalization performance > <ref:2602.05451#pg19> By leveraging the powerful capacity of deep neural networks (DNNs) to extract non-linear features, this framework can unbiasedly pinpoint cell populations most responsive to mechanical stimulation from high-dimensional single-cell data without reliance on prior knowledge > <ref:2602.05451#pg21>

The researchers demonstrated a Somatic–Visceral Force–Immune Axis orchestrated by CPTCs, showing that CPTCs in the subcutaneous fascia act as primary responders to acupuncture mechanical stimulation, sensing stress via an AP-1/Hsp70 axis and secreting Wnt2-loaded extracellular vesicles (EVs) into the circulation > <ref:2602.05451#pg22> This systemic elevation of Wnt signals induces a transcriptional resonance in distal colonic CPTCs, which subsequently reprograms the colonic immune microenvironment and activates the Wnt/β-catenin/Myc regenerative program in epithelial cells to repair injury caused by bacterial colitis > <ref:2602.05451#pg22>

Key Findings on Mechanosensing and Signaling

The CARSS framework revealed that Fascial CPTCs emerged as the top-ranking responder, achieving a training accuracy of 99.86% and a remarkable generalization accuracy of 86.32% in complex mixed datasets (Figure 2A) > <ref:2602.05451#pg18> This high Generalization Index (G-Index < 1.0) identifies them as the most robust cellular biosensors for acupuncture stimulation > <ref:2602.05451#pg18> Mechanistic dissection using SHAP analysis identified the AP-1/Hsp70 axis as the core mechanotransduction pathway in CPTCs, with immediate-early genes Jun and Fos being core predictive features > <ref:2602.05451#pg21>

Systemic Communication and Transcriptional Resonance

The study observed a striking transcriptional resonance—despite their anatomical distance, they shared a highly conserved transcriptional trajectory and Wnt signaling profile (Figure 6D, see also Figure S7A, B) > <ref:2602.05451#pg22> This systemic synchronization was quantitatively confirmed using single-cell data from fascial CPTCs and colonic CPTCs only, showing a strong positive correlation (R = 0.75) in the transcriptional fold-changes of key mechanosensitive genes between the two CPTC populations > <ref:2602.05451#pg22> Notably, the mechanical alarmin Il33 and the regenerative ligand Wnt2 exhibited a lock-step upregulation pattern in both fascial and colonic CPTCs (Figure S7C) > <ref:2602.05451#pg22>

Functional Outcomes of Somatic Mechanotherapy

The therapeutic efficacy was confirmed at the macroscopic level, where acupuncture treatment significantly reversed pathological changes in rat colon tissues, restoring crypt morphology and reducing inflammatory infiltration > <ref:2602.05451#pg4> At single-cell resolution, mechanotherapy promoted epithelial proliferation by consistently increasing the proportion of cells in the G2/M phase across multiple subsets including enterocytes and goblet cells > <ref:2602.05451#pg4> Furthermore, the CPTC secretome was shown to directly rescue epithelial barrier integrity by suppressing TLR4 and IL6 expression while robustly upregulating the Wnt target genes CTNNB1 (β-catenin) and MYC in the epithelial cells > <ref:2602.05451#pg4>

Conclusion on Communication

The findings suggest that acupuncture induces a systemic healing state where fascial and colonic CPTCs operate in transcriptional unison to restore tissue homeostasis, with serum Wnt2 acting as a potential quantifiable biomarker for acupuncture intervention efficacy against bacterial dysentery > <ref:2602.05451#pg4> This mechanism demonstrates that peripheral mechanical stress is faithfully translated into visceral regenerative signals through a systemic microenvironmental reprogramming characterized primarily by enhanced reparative immunomodulatory signaling > <ref:2602.05451#pg4>

How it works

The study established a complete chain of events: (1) subcutaneous fascial CPTCs immediately sense and respond to mechanical stress through the AP-1 axis; (2) acupuncture intervention signals are transduced into systemic upregulation of Wnt signaling; (3) subcutaneous fascial CPTCs and colonic CPTCs undergo resonance, while strengthening the Wnt signaling circuit, targeting intestinal epithelium and other intestinal microenvironment cells to mitigate the impact of intestinal inflammatory storms and promote the overall transition of the intestine toward an immune-repair mode > <ref:2602.05451#pg4>

The in vitro co-culture experiments demonstrated that merely introducing the subcutaneous fascial CPTC secretome as a microenvironmental component was sufficient to reduce Salmonella proliferation capacity and dramatically reduce infection-induced apoptosis, while simultaneously activating the Wnt/β-catenin/Myc pathway in intestinal epithelial cells > <ref:2602.05451#pg4> This result aligns perfectly with the latest research demonstrating that peripheral blood Wnt2 is an acute-phase anti-apoptotic factor > <ref:2602.05451#pg4>

The mechanism for long-distance transport of Wnt proteins is addressed by TEM evidence showing that EVs enriched around CPTCs suggest that acupuncture-mobilized Wnt2 does not exist as free protein but is encapsulated within lipid bilayers, conferring endocrine hormone-like remote homing capability > <ref:2602.05451#pg4> This vesicular packaging mechanism protects Wnt proteins from degradation and suggests that serum Wnt2 acts as a convergence pool for this signal, marking the organism's entry into a pro-regenerative state > <ref:2602.05451#pg4>

The study concludes that the therapeutic efficacy of acupuncture intervention can be summarized at the systems biology level as a systemic microenvironmental reprogramming characterized primarily by enhanced reparative immunomodulatory signaling > <ref:2602.05451#pg4> This approach provides a reusable research paradigm for acupuncture research based on systems biology knowledge > <ref:2602.05451#pg4>

How it works

The CARSS deep learning framework was designed with adaptive MLP models and a two-phase grid search to systematically evaluate 21,600 parameter combinations for each cell type > <ref:2602.05451#pg18> The model evaluation utilized a dual strategy of self-matrix testing and generalization testing, calculating the Generalization Index (G-Index = Generalization Accuracy / Self-Test Accuracy) to quantify the robustness of mechanosensitive signatures > <ref:2602.05451#pg21>

The interpretation of molecular features was performed using SHAP (SHapley Additive exPlanations) analysis, which identified genes such as Hspa1b and Jun as core predictive features in fascial CPTCs, establishing the AP-1/Hsp70 axis as the primary mechanotransduction mechanism > <ref:2602.05451#pg21> This echoes previous biomechanical studies showing that mechanical force can directly induce rapid AP-1 phosphorylation and nuclear translocation, initiating downstream gene transcription > <ref:2602.05451#pg4>

The analysis also revealed a hierarchical response among fascial cell types, where Fascial CPTCs exhibited the strongest positive enrichment in the Response to mechanical stimulus pathway (GO:0009612), while other responsive cells showed nonsignificant negative enrichment for this pathway > <ref:2602.05451#pg4> This highlights that macrophages and fibroblasts may undergo reprogramming of their other functions by acupuncture rather than directly responding to mechanical stimulation > <ref:2602.05451#pg4>

The study utilized CellChat analysis to predict a robust Wnt signaling flow from fascial CPTCs to colonic CPTCs, mediated by Wnt2/Wnt10b ligands, which was validated by observing significantly enhanced Wnt signaling pathway in the acupuncture group > <ref:2602.05451#pg22> This confirms that the visceral regenerative program acts as a direct scalar echo of the peripheral mechanical response > <ref:2602.

1. Bold header: Cellular Response Scoring System (CARSS) Integration

The improved system can perform unsupervised identification of mechanosensitive cell populations by leveraging generalization performance as a machine learning problem, allowing it to unbiasedly pinpoint cell populations most responsive to mechanical stimulation from high-dimensional single-cell data without reliance on prior knowledge.

2. Bold header: Mechanistic Axis Decoding via SHAP Analysis

The system can decode the core molecular features driving cellular state transitions by using SHAP (SHapley Additive exPlanations) analysis, specifically identifying genes like Jun and Fos as the immediate-early response signature of cells responding to mechanical strain.

3. Bold header: Cross-Tissue Transcriptional Resonance Mapping

The improved system can identify systemic communication between distant tissues by analyzing the transcriptional resonance between fascial and colonic CPTCs, confirming that they share a highly conserved transcriptional trajectory and Wnt signaling profile.

4. Bold header: Systemic Biomarker Prediction

The system can predict acupuncture intervention efficacy by identifying serum Wnt2 as a potential quantifiable biomarker, based on the finding that the acupuncture-induced elevation of serum Wnt2 protein concentration suggests a systemic pro-regenerative state.

5. Bold header: Causal Intervention Simulation

The system can simulate therapeutic outcomes by integrating CPTC secretome data to predict barrier rescue, as it demonstrates that the CPTC secretome—whether acting locally or systemically via Wnt2-loaded EVs—is sufficient to activate epithelial regeneration and restore barrier integrity.

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+PDGFRα+ 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-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 somatic mechanical stimulation into systemic regenerative microenvironments.