Osteoarthritis (OA) is a debilitating chronic joint disease affecting large populations of patients, especially the elderly. The pathological mechanisms of OA are currently unknown. Multiple risk factors are involved in OA development. Among these risk factors, alterations of mechanical loading in the joint leading to changes in biological signaling pathways have been known as a key event in OA development. The importance of AMPK-β-catenin-Runx2 signaling in the initiation and progression of OA has been recognized in recent years. In this review, we discuss the recent progress in understanding the role of this signaling pathway and the underlying interaction mechanisms during OA development. We also discuss the drug development aiming to target this signaling pathway for OA treatment.
Nanotechnology has made a significant impact on the development of nanomedicine. Nonviral vectors have been attracting more attention for the advantage of biosafety in gene delivery. Polyethylenimine (PEI)-conjugated chitosan (chitosan-g-PEI) emerged as a promising nonviral vector and has been demonstrated in many tumor cells. However, there is a lack of study focused on the behavior of this vector in stem cells which hold great potential in regenerative medicine. Therefore, in this study, in vitro gene delivering effect of chitosan-g-PEI was investigated in bone marrow stem cells. pIRES2-ZsGreen1-hBMP2 dual expression plasmid containing both the ZsGreen1 GFP reporter gene and the BMP2 functional gene was constructed for monitoring the transgene expression level. Chitosan-g-PEI-mediated gene transfer showed 17.2% of transfection efficiency and more than 80% of cell viability in stem cells. These values were higher than that of PEI. The expression of the delivered BMP2 gene in stem cells enhanced the osteogenic differentiation. These results demonstrated that chitosan-g-PEI is capable of applying in delivering gene to stem cells and providing potential applications in stem cell-based gene therapy.
A high strength hydrogel was fabricated by one-step copolymerization of dipole–dipole interaction-containing monomer, acrylonitrile, super-hydrophilic comonomer, 2-methacryloyloxyethyl phosphorylcholine and crosslinker, polyethylene glycol diacrylate (Mn = 575, PEGDA575). This dipole–dipole reinforced (DDR) hydrogel demonstrated intriguing combinations of properties such as withstanding several MPa tensile stress, tens of MPa compressive strength, excellent fatigue resistance and no yielding during tensile tests. The equilibrium water content and transparency of DDR hydrogels could be tuned by varying monomer concentration and monomer ratio. The gels exhibited low cytotoxicity and antifouling characteristic. Biodegradable high strength hydrogel could also be constructed by merely replacing PEGDA575 with bioreducible crosslinker. The method reported here offers a general strategy to design biocompatible high-strength hydrogels for tissue engineering scaffolds by copolymerizing monomer containing dipole–dipole pairing with other hydrophilic monomer.
NeoTCR-P1 is a personalized autologous T cell therapy for treatment of patients with solid tumors. Neoantigen-specific T cell receptors (neoTCRs) were isolated from the patients' own circulating CD8 T cells using the imPACT Isolation Technology®, followed by non-viral precision genome engineering into an autologous apheresis product for infusion back into the patient.
Methods
This phase 1 trial is a first-in-human, multi-center, dose-escalation study to evaluate the safety, tolerability, and manufacturing feasibility of NeoTCR-P1 alone or in combination with IL-2 in solid tumors. Patients with TCRs identified at screening and meeting eligibility criteria underwent apheresis to manufacture personalized NeoTCR-P1 cell product. Lymphodepleted patients received a single dose of up-to-three distinct NeoTCR cell products at dose levels of 0.4, 1.2, or 4×109 NeoTCR-edited T cells. Pre- and post-treatment blood and biopsy samples were collected to evaluate NeoTCR-P1 pharmacokinetics, tumor trafficking, signs of T cell engagement or potential mechanisms of resistance.
Results
Sixteen patients were infused with NeoTCR-P1 T cells including patients with MSS-colorectal cancer (11), breast cancer (2), ovarian cancer (1), melanoma (1), or non-small cell lung cancer (1). Four of the sixteen patients were treated with NeoTCR-P1 + IL-2. Two patients experienced toxicities associated with NeoTCR-P1 cell infusions: a grade 1 CRS and a grade 2 ICANS. Five patients had stable disease as their best response at their first tumor assessment (day 28). NeoTCR+ T cells detected in the peripheral blood had an average peak of 3.6% (range 0.9-7.3%) for DL1, 11.7% (7.7-20.8%) for DL2, and 19.8% (12.0-37.3%) for DL3. Increases in NeoTCR T cells were observed at higher dose levels, stronger lymphodepletion, or higher gene editing rates of the infused product. Eight post-infusion biopsies were available for sequencing and imaging analysis; 17 of 22 neoTCR-T cells were detected in post-infusion biopsies with 12 neoTCRs among the top 4% of CDR3 sequences detected. The targeted neoantigens were detected in 7 of 8 post-treatment biopsies (15 of 22 targets), and personalized ctDNA confirmed targeting of a predicted sub-clonal mutation. An APOBEC signature and HLA-LOH were identified as potential mechanisms of resistance. By single-cell, spatial molecular imaging, neoTCR-T cells were visualized in post-treatment biopsies and found to differentially express potential markers of engagement.
Conclusions
This study demonstrates the feasibility of isolating and manufacturing NeoTCR-T cells using non-viral precision genome engineering, the safety of infusing up-to-three gene edited NeoTCR-T cell products, and T cell persistence and trafficking to a variety of solid tumors.
Trial Registration
NCT03970382
Ethics Approval
Ethics approvals have been obtained from each clinical site enrolling patients: City of Hope, Duarte California; University of California Los Angeles, Los Angeles California; University of California, Irvine Medical Center, Orange, California; University of California, Davis, Sacramento California; University of California, San Francisco, San Francisco California; Northwestern University Medical Center, Chicago Illinois; Memorial Sloan Kettering Cancer Center, New York, New York; Tennessee Oncology, Nashville, Tennessee; and Fred Hutchinson Cancer Research Center, Seattle, Washington.
Abstract Large segment bone defects pose a significant challenge in the field of orthopedic surgery, requiring effective and innovative approaches for restoration. However, many existing scaffolds are bioinert and do not support crucial processes such as cell adhesion, proliferation, and vascularization. In this study, a dual‐bionic 3D printing bredigite scaffold is developed, featuring a combination of physical structure and bioactive functions. Specifically, the structure‐mimetic scaffold has an isotropic single‐cell structure suitable for defects with varying load‐bearing requirements and allowing the ingrowth of vessels and bone. Meanwhile, an extracellular matrix peptide‐mimetic β‐amino acid polymer DM 50 CO 50 and deferoxamine are modified onto the scaffold simultaneously to promote the adhesion of bone marrow mesenchymal stem cells and vascularization. The dual‐bionic scaffolds demonstrate outstanding osteogenic and angiogenic properties in a rat model with large segment bone defects to promote bone restoration, implying a promising strategy in designing scaffolds to promote osteoconductivity and angiogenesis for large segment bone restoration.
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