Surface-modified injectable poly(ethylene-glycol) diacrylate-based cryogels for localized gene delivery.

Lentiviral transduction is widely used in research, has shown promise in clinical trials involving gene therapy and has been approved for CAR-T cell immunotherapy. However, most modifications are doneex vivoand rely on systemic administration of large numbers of transduced cells for clinical applications. A novel approach utilizingin situbiomaterial-based gene delivery can reduce off-target side effects while enhancing effectiveness of the manipulation process. In this study, poly(ethylene glycol) diacrylate (PEGDA)-based scaffolds were developed to enablein situlentivirus-mediated transduction. Compared to other widely popular biomaterials, PEGDA stands out due to its robustness and cost-effectiveness. These scaffolds, prepared via cryogelation, are capable of flowing through surgical needles in bothin vitroandin vivoconditions, and promptly regain their original shape. Modification with poly(L-lysine) (PLL) enables lentivirus immobilization while interconnected macroporous structure allows cell infiltration into these matrices, thereby facilitating cell-virus interaction over a large surface area for efficient transduction. Notably, these preformed injectable scaffolds demonstrate hemocompatibility, cell viability and minimally inflammatory response as shown by ourin vitroandin vivostudies involving histology and immunophenotyping of infiltrating cells. This study marks the first instance of using preformed injectable scaffolds for delivery of lentivectors, which offers a non-invasive and localized approach for delivery of factors enablingin situlentiviral transduction suitable for both tissue engineering and immunotherapeutic applications.

Acellular scaffold-based approach for in situ genetic engineering of host T-cells in solid tumor immunotherapy.

BACKGROUND: Targeted T-cell therapy has emerged as a promising strategy for the treatment of hematological malignancies. However, its application to solid tumors presents significant challenges due to the limited accessibility and heterogeneity. Localized delivery of tumor-specific T-cells using biomaterials has shown promise, however, procedures required for genetic modification and generation of a sufficient number of tumor-specific T-cells ex vivo remain major obstacles due to cost and time constraints. METHODS: Polyethylene glycol (PEG)-based three-dimensional (3D) scaffolds were developed and conjugated with positively charged poly-L-lysine (PLL) using carbamide chemistry for efficient loading of lentiviruses (LVs) carrying tumor antigen-specific T-cell receptors (TCRs). The physical and biological properties of the scaffold were extensively characterized. Further, the scaffold loaded with OVA-TCR LVs was implanted in B16F10 cells expressing ovalbumin (B16-OVA) tumor model to evaluate the anti-tumor response and the presence of transduced T-cells. RESULTS: Our findings demonstrate that the scaffolds do not induce any systemic inflammation upon subcutaneous implantation and effectively recruit T-cells to the site. In B16-OVA melanoma tumor-bearing mice, the scaffolds efficiently transduce host T-cells with OVA-specific TCRs. These genetically modified T-cells exhibit homing capability towards the tumor and secondary lymphoid organs, resulting in a significant reduction of tumor size and systemic increase in anti-tumor cytokines. Immune cell profiling revealed a significantly high percentage of transduced T-cells and a notable reduction in suppressor immune cells within the tumors of mice implanted with these scaffolds. CONCLUSION: Our scaffold-based T-cell therapy presents an innovative in situ localized approach for programming T-cells to target solid tumors. This approach offers a viable alternative to in vitro manipulation of T-cells, circumventing the need for large-scale in vitro generation and culture of tumor-specific T-cells. It offers an off-the-shelf alternative that facilitates the use of host cells instead of allogeneic cells, thereby, overcoming a major hurdle.

Novel combination of bioactive agents in bilayered dermal patches provides superior wound healing.

In present study, multifunctional bilayered dermal patches with antibacterial, antioxidant and anti-inflammatory properties were developed using solvent casting or electrospinning methods and compared for performance. Top layer was made up of polycaprolactone (PCL) and chitosan (CS) while bottom layer was made of polyvinyl alcohol (PVA) with curcumin nanoparticles and soluble eggshell membrane protein (SESM) as the wound healing agents. Curcumin nanoparticles showed reduction in the production of reactive oxygen species (ROS) and inflammatory cytokines and markers in mice T cells or human macrophages, confirming their antioxidant and anti-inflammatory properties while SESM improved migration of human adult dermal fibroblasts, suggesting its contribution to wound healing. The dermal patches were hemocompatible and antibacterial and also provided adequate absorption of wound exudates, support and components required for recruitment of cells and deposition of extracellular matrix to enable superior wound healing than its commercial counterpart in a full thickness excision wound model in rats.

Immunomodulation via macrophages to fight solid tumor malignancies.

The paper 'A C-Terminal Fragment of Adhesion Protein Fibulin-7 Inhibits Growth of Murine Breast Tumor by Regulating Macrophage Reprogramming' by Chakraborty et al. highlights that Fbln7-C could be explored as a potential immunomodulatory agent against various solid cancers and have shown its abilities to regulate tumor microenvironment reprogramming of TAMs in a breast cancer model. Fbln7, which is a secreted glycoprotein, has been shown to be anti-angiogenic and has an immunomodulatory role regulating various functional properties of monocytes, macrophages, and neutrophils, thereby influencing inflammation. In this study, the authors have shown that in a murine breast tumor model, intravenous administration of Fbln7-C significantly reduces the size of tumors via macrophage reprogramming. Comment on: https://doi.org/10.1111/febs.15333.

Glycated collagen - a 3D matrix system to study pathological cell behavior.

A hyperglycemic condition like diabetes in patients renders them with an increased risk of developing breast cancer. Hyperglycemia and ageing increase the non-enzymatic glycosylation (glycation) of nearly all proteins in our body including collagen type I, which is an important extracellular matrix (ECM) component. This results in the formation of advanced glycated end products (AGEs), which can form covalent crosslinks in collagen fibers and change the overall architecture and stiffness of the matrix. In this study we have used MDA-MB-231 breast cancer cells to study the interaction of tumor cells with glycated collagen and have explored the role of matrix architecture and RAGE-mediated signaling in cellular behavior. We mimicked the non-enzymatic glycation of protein by treating collagen I with glucose or ribose and found that crosslinking due to AGEs induces collagen fiber bundling and an increase in pore size and stiffness of the matrix. We also observed that AGE formation triggers AGE-RAGE signaling playing a role in the morphology and migration of cells. Furthermore, our study suggests an interplay of the pore size of the collagen matrix and RAGE mediated signaling in 3D invasion of cells and our findings demonstrate that the effect of the AGE-RAGE interaction is more pronounced than that of an altered matrix architecture. This study has helped us develop a 3D system using glycated collagen to study the effects of pathological conditions such as diabetes on extracellular matrix proteins, which may have downstream effects on cell behavior and dysfunction.