Enhancing MSC Therapy for Acute GVHD via CRISPLD2 Modulation

Introduction to Acute Graft-Versus-Host Disease and MSC Therapy

Acute Graft-Versus-Host Disease (aGVHD) is a severe complication that arises after allogeneic hematopoietic stem cell transplantation (allo-HSCT), where the donor’s immune cells attack the recipient’s tissues. It poses significant challenges in treatment and often limits the success of transplantation strategies (Xu et al., 2025). The incidence of aGVHD ranges from 30% to 50% in individuals undergoing allo-HSCT, making it a critical area of research in transplantation medicine (Xu et al., 2025).

Mesenchymal stem cells (MSCs), particularly human umbilical cord-derived MSCs (HUC-MSCs), have emerged as a promising therapeutic option for preventing and treating aGVHD due to their immunosuppressive properties and ability to modulate immune responses (Xu et al., 2025). MSCs can inhibit T-cell proliferation and promote the secretion of anti-inflammatory cytokines, such as IL-10, enhancing their therapeutic efficacy in aGVHD (Xu et al., 2025). However, the heterogeneity of MSCs can negatively influence their outcomes, necessitating strategies to enhance their effectiveness.

Significance of CRISPLD2 in Human Umbilical Cord-Derived MSCs

Cysteine-rich secretory protein LCCL domain-containing 2 (CRISPLD2) has been identified as a crucial marker for immunosuppressive HUC-MSCs subsets (Xu et al., 2025). Recent studies have shown that CRISPLD2 expression correlates with the immunomodulatory capabilities of MSCs, suggesting that enhancing its expression could improve the therapeutic potential of HUC-MSCs in treating aGVHD (Xu et al., 2025).

Single-cell transcriptomic analyses have revealed that CRISPLD2 is heterogeneously expressed among HUC-MSCs, highlighting its role in distinguishing functionally distinct subpopulations (Xu et al., 2025). Importantly, CRISPLD2 overexpression in HUC-MSCs has been shown to enhance their immunosuppressive functions, potentially through inhibition of the P2Y11 receptor signaling pathway, which mediates calcium signaling in MSCs (Xu et al., 2025).

Mechanisms of CRISPLD2 in Immunosuppression and Mitochondrial Health

CRISPLD2 overexpression in HUC-MSCs leads to significant changes in gene expression profiles associated with immune modulation and cellular signaling. Functional assays indicate that CRISPLD2 enhances the secretion of IL-10 and indoleamine 2,3-dioxygenase (IDO), both of which play vital roles in T-cell suppression and immune regulation (Xu et al., 2025). Furthermore, CRISPLD2 seems to mitigate the detrimental effects of extracellular ATP and lipopolysaccharides (LPS) on HUC-MSC function, preserving their stemness and immunosuppressive capabilities (Xu et al., 2025).

In addition to its immunomodulatory effects, CRISPLD2 is associated with mitochondrial health in HUC-MSCs. Mitochondria are crucial for cellular metabolism and energy production, and their dysfunction can lead to impaired MSC function. The downregulation of P2Y11, a purinergic receptor that mediates cellular responses to ATP, was observed in CRISPLD2-overexpressing HUC-MSCs, indicating that CRISPLD2 may play a protective role against mitochondrial damage induced by ATP and inflammatory signals (Xu et al., 2025).

Impact of CRISPLD2 Overexpression on T-Cell Function and Cytotoxicity

The overexpression of CRISPLD2 in HUC-MSCs significantly enhances their ability to suppress T-cell proliferation, as demonstrated in coculture experiments with both murine and human T cells (Xu et al., 2025). In these studies, T cells cultured with CRISPLD2-modified HUC-MSCs exhibited lower proliferation rates compared to those cultured with control HUC-MSCs, underscoring the enhanced immunosuppressive function of the modified cells.

Moreover, IL-10 production was markedly increased in the culture media of CRISPLD2-overexpressing HUC-MSCs. This increase in IL-10 levels is critical for the inhibition of T-cell activity, providing a mechanistic understanding of how CRISPLD2 enhances the immunosuppressive capabilities of HUC-MSCs (Xu et al., 2025).

Table 1: Comparison of T-Cell Proliferation Rates

Future Directions for CRISPLD2-Enhanced MSC Therapies in GVHD

The findings from recent research highlight the potential for CRISPLD2 modulation in HUC-MSCs to improve outcomes in patients at risk of aGVHD. Future studies should focus on the following areas:

1. Clinical Trials: Conducting clinical trials to evaluate the safety and efficacy of CRISPLD2-modified HUC-MSCs in patients undergoing allo-HSCT.
2. Mechanistic Studies: Further elucidating the mechanisms through which CRISPLD2 influences mitochondrial health and T-cell interactions to optimize therapeutic strategies.
3. Long-term Outcomes: Investigating the long-term effects of CRISPLD2 overexpression on MSC function and patient outcomes, particularly concerning the balance between immunosuppression and GVL activity.

FAQ Section

What is aGVHD and why is it significant?

Acute Graft-Versus-Host Disease (aGVHD) is a serious complication following allogeneic stem cell transplantation where donor immune cells attack the recipient’s tissues. Its significance lies in its high incidence and the potential to limit the effectiveness of transplant procedures.

How do HUC-MSCs help in treating aGVHD?

HUC-MSCs possess immunosuppressive properties that can inhibit T-cell proliferation and modulate immune responses, thereby reducing the severity of aGVHD.

What role does CRISPLD2 play in HUC-MSCs?

CRISPLD2 is a marker for immunosuppressive subpopulations of HUC-MSCs. Its overexpression enhances the immunosuppressive functions of these cells, potentially improving their efficacy in aGVHD treatment.

Are there any risks associated with using CRISPLD2-modified MSCs?

While preliminary results are promising, further research is necessary to evaluate the long-term safety and potential off-target effects of CRISPLD2 modulation in MSCs.

References

1. Xu, Q., Wang, R., Sui, K., Xu, Y., Zhou, Y., He, Y., Hu, Z., Wang, Q., Xie, X., Wang, X., Yang, S., Zeng, L., Zhong, J. F., & Wang, Z. (2025). Enhance the therapeutic efficacy of human umbilical cord-derived mesenchymal stem cells in prevention of acute graft-versus-host disease through CRISPLD2 modulation. Stem Cell Research & Therapy, 1757-6512. https://doi.org/10.1186/s13287-025-04321-6

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