Mesenchymal stromal cells (MSCs) are among the best-characterized cell types in regenerative medicine. They can be readily expanded in culture and differentiated into various tissues and cells, while also exhibiting immunomodulatory characteristics. However, MSCs have limited therapeutic efficacy because they have low survival rates, are heterogeneous, and lose their ability to function after transplantation. New advances in genome editing with CRISPR-Cas9 have opened the door to targeted genetic manipulation of MSCs, ushering in an era of improved regenerative potential, particularly for bone and cartilage repair. This review focuses on recent advances in using CRISPR-modified MSCs for musculoskeletal regeneration, with particular emphasis on promoting osteogenesis, chondrogenesis, angiogenesis, immunomodulation, and extracellular matrix remodeling. Preliminary studies have identified several key targets, including the transcription factor RUNX2, which is associated with positive differentiation outcomes; CXCR4, which is linked to tissue-specific homing; IL-10 as an anti-inflammatory target; and PD-L1 as an immunomodulatory target with promising therapeutic potential. Furthermore, cell-free therapeutic approaches, based on the secretome and extracellular vesicles (EVs) engineered with MSCs, have gained increasing attention. Recently, studies have also focused on enhancing tissue regeneration by using genetically engineered MSCs in biomaterials and hydrogel systems, as well as scaffold-based systems. Although preclinical results are encouraging, challenges such as off-target effects, genomic stability, delivery efficiency, manufacturing standardization, and long-term safety concerns still hinder clinical translation. More importantly, no clinical trials have registered the use of engineered MSCs using the CRISPR for cartilage and/or bone regeneration, indicating a disconnect between experimental progress and clinical use. Overall, this CRISPR-based “genetic modification” approach to MSCs is a promising strategy to enhance their therapeutic potential and develop the next generation of regenerative treatments for skeletal diseases.
Photo-oxidative stress is a key contributor to vision loss in older adults and involves progressive dysfunction and loss of photoreceptors and retinal pigment epithelial (RPE) cells. Effective therapies that directly preserve outer retinal cell survival under such stress remain limited. Ascl1 is a basic helix-loop-helix transcription factor best known for neurogenesis and cellular plasticity and has been widely studied in the retina for its ability to reprogram Müller glia. Here we asked whether Ascl1 gene augmentation can also confer cytoprotection in the adult outer retina under acute degenerative stress. We used a mouse model of blue light-induced retinal injury to impose photo-oxidative stress on the outer retina and RPE, and tested the protective effect of Ascl1 gene augmentation using AAV8-CAG-Ascl1-GFP (with AAV8-CAG-GFP as the control). Retinal structure and function were assessed using immunostaining, behavioral tests, and electroretinography. To explore the underlying mechanisms, we further performed a dual-luciferase reporter assay, bulk RNA sequencing, qPCR, and western blotting. Ascl1 overexpression was detected in the outer retina and RPE, where it mitigated blue light-induced degeneration, resulting in the partial preservation of outer retinal structure and retinal function (ERG b-wave responses) and light/dark preference behavior. Consistent with an anti-apoptotic effect, Ascl1 markedly reduced TUNEL-positive cells in the ONL and was accompanied by decreased cleaved Caspase-8 and BAX and increased Bcl2. Dual-luciferase assays confirmed that Ascl1 directly activates the Bcl2 promoter. In parallel, Ascl1 increased PI3K and AKT phosphorylation, suggesting engagement of pro-survival signaling. Injury-associated glial/microglial activation and RPE junctional disruption were also attenuated in Ascl1-treated eyes. These data support a cytoprotective dimension of Ascl1 augmentation in addition to its reprogramming-associated role and demonstrate that AAV-based Ascl1 augmentation can enhance retinal stress resilience following acute photo-oxidative injury, motivating further evaluation in chronic, age-associated models with dose/safety optimization and pathway-level validation.
Colorectal cancer (CRC) constitutes a major global health burden, and its high mortality is closely associated with tumor immune evasion. Metabolic reprogramming of regulatory T cells (Tregs) in the tumor microenvironment (TME) is a key mechanism that maintains their immunosuppressive phenotype. Key metabolic pathways, including glycolysis, fatty acid oxidation (FAO), amino acid metabolism, the pentose phosphate pathway (PPP), and the PI3K-AKT-mTOR signaling cascade, coordinately enable Tregs to adapt to the hypoxic and nutrient-deprived TME and preserve their suppressive functions. The gut microbiome and its metabolites, such as short-chain fatty acids (SCFAs) and bile acids (BAs), exert distinct regulatory effects on Treg metabolism within CRC. Targeting these metabolic dependencies offers innovative therapeutic strategies to attenuate Treg-mediated immunosuppression. This review systematically summarizes the metabolic adaptation of Tregs in the CRC TME, and delineates the regulatory roles of the gut microbiome. It further evaluates the synergistic potential of combining metabolic interventions with current immunotherapies, including immune checkpoint inhibitors (ICIs) and adoptive cell therapies (ACTs). Finally, we outline emerging translational opportunities and propose optimized clinical trial designs to address existing challenges and chart future research directions. Clinical trial numberNot applicable.
Regional homogeneity (ReHo) reflects local synchronization of resting-state brain activity, but the structural and molecular correlates of ReHo across the Alzheimer’s disease (AD) continuum remain unclear. We included 421 cognitively normal participants, 226 patients with mild cognitive impairment, and 68 patients with AD. T1-weighted MRI, diffusion MRI, and resting-state functional MRI were used to quantify cortical thickness, regional streamline count, structure-function coupling (SFC), and ReHo. Disease abnormality indices were constructed across diagnostic stages using ordinal regression. Generalized linear models and repeated cross-validation were used to assess the associations of structural and coupling measures with regional ReHo. Residual ReHo after adjustment for cortical thickness, regional streamline count, and SFC was spatially associated with gene-expression maps using partial least squares regression, followed by pathway, protein interaction, and cell-type enrichment analyses. ReHo, cortical thickness, regional streamline count, and SFC showed stage-dependent and spatially heterogeneous abnormalities. Disease abnormality indices of ReHo, cortical thickness, regional streamline count, and SFC were associated with broad cognitive and functional impairment. ReHo was consistently associated with cortical thickness (β, −0.210 to −0.129; all p < 0.001), regional streamline count (β, 0.001 to 0.059; all p < 0.05), and SFC (β, 0.166 to 0.197; all p < 0.001), which jointly explained regional ReHo variance (R2, 0.346 to 0.371). Residual ReHo was linked to cortical gene-expression gradients enriched for neurovascular signaling, energy metabolism, proteostasis, and stage-specific astrocytic, microglial, excitatory neuronal, and inhibitory neuronal processes. ReHo alterations across the AD continuum are associated with structural, functional, and molecular features. This work advances our understanding of regional functional alterations across the AD continuum and helps inform the development of targeted intervention strategies.
