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Biology subjects

Francis, M. P.

Publications and source records attributed to Francis, M. P..

3 recordsLinked to original sources

High Precision Peptide-MHC Class I CARs Enhance In Vivo Killing ofTargeted CD8 T Cells to Prevent Type 1 Diabetes in Novel RIP-mOVA Model

Progress has been made to address refractory T cell-mediated diseases with chimeric antigen receptor (CAR) T cell platforms, with the need more specifically target pathogenic cells being addressed by peptide-MHC (pMHC) CAR T cells. However, progress to date has largely been limited to peptide-MHC Class II (pMHC-II) CARs for the targeting of autoreactive CD4 T cells in multiple models. Here, we develop the first single molecule peptide-MHC Class I-{beta}2 microglobulin CAR platform to demonstrate the proof of concept for depletion of antigen-specific CD8 T cells and compare the in vitro and in vivo functionality to a traditional scFv-based CAR that also targets the TCR. We demonstrate that while the scFv CAR is more rapidly and robustly activated by target CD8 T cells in vitro, the greater precision offered by pMHC-I CARs results in greater efficacy in multiple in vivo models. Additionally, we developed a novel adjuvanted ovalbumin vaccination with adoptive transfer model to induce type 1 diabetes (T1D) in RIP-mOVA mice using a polyclonal, polyepitopic T cell population. We demonstrate that our pMHC-I platform depletes SIINFEKL-reactive T cells and that control of the immune response to the dominant SIINFEKL epitope can control a broader antigenic response.

immunology↗

Assembled Cell-Decorated Collagen (AC-DC) bioprinted implants mimic musculoskeletal tissue properties and promote functional recovery

Musculoskeletal tissue injuries, including the damage and rupture of ligaments and tendons, and volumetric muscle loss (VML), are exceptionally commonplace and often lead to permanent disability and deformation. We developed an advanced biomanufacturing platform producing cellularized collagen microfiber implants to facilitate functional repair and regeneration of musculoskeletal soft tissues. This Assembled Cell-Decorated Collagen (AC-DC) bioprinting process rapidly and reproducibly forms 3D implants using clinically relevant cells and strong, microfluidic extruded collagen fibers. Quantitative analysis showed that the directionality and distribution of cells throughout AC-DC implants mimic the cellular properties of native musculoskeletal tissue. AC-DC bioprinted implants further approximate or exceed the strength and stiffness of human tendons and ligaments and exceeded the properties of commonplace collagen hydrogels by orders of magnitude. The regenerative potential of AC-DC implants was also assessed in vivo in a rodent VML model. A critically sized muscle injury in the hindlimb was created and repaired, and limb torque generation potential was measured over 12 weeks. Both acellular and cellular implants were found to promote functional recovery compared to the unrepaired group, with AC-DC implants containing therapeutic muscle progenitor cells promoting the highest degree of recovery. Histological analysis and automated image processing of explanted muscle cross-sections revealed increased total muscle fiber count, median muscle fiber size, and increased cellularization for injuries repaired with cellularized implants. These studies introduce the tremendous potential of an advanced bioprinting method for generating tissue analogs with near-native biological and biomechanical properties with the potential to repair numerous challenging musculoskeletal injuries. One Sentence SummaryBioprinted collagen microfiber-based implants mimic musculoskeletal tissue properties in vitro and promote functional recovery in vivo.

bioengineering↗

Direct Crystal Formation from Micronized Bone and Lactic Acid: The Writing on the Wall for Calcium-Containing Crystal Pathogenesis in Osteoarthritis?\"

IntroductionPathological calcium-containing crystals accumulating in the joints, synovial fluid, and soft tissues are noted in most elderly patients, yet arthritic crystal formation remains idiopathic. Interestingly, elevated lactic acid and bone erosion are frequently among the comorbidities and clinical features of patients with highest incidence of crystal arthropathies. This work shows that bone particulates (modeling bone erosion) dissolve in lactic acid and directly generate crystals, possibly presenting a mechanism for crystal accumulation in osteoarthritis.\n\nMethods and ResultsMicronized human bone (average particle size of 160m x 79m) completely dissolved in lactic acid in 48 hours, and in synovial fluid with 500mM lactic acid in 5 days, generating birefringent rhomboid and rod-shaped crystals. SEM analysis with energy dispersive x-ray spectroscopy of these crystals showed average dimensions of around 2m x 40m, which contained oxygen, calcium and phosphorous at 8.64:1.85:1. Raman spectroscopy of the generated crystals further showed 910/cm and 1049/cm peaks, aligning with calcium oxalate monohydrate and calcium pyrophosphate, respectively.\n\nConclusionsThis work shows that lactic acid and micronized mineralized bone together directly generate calcium-containing crystals. These observations may provide insights into the elusive etiology of arthritis with crystal involvement, possibly indicating lactic acid as a clinical target for treatment.

pathology↗