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Principal Investigator: Donald G Phinney
Organization: UNIVERSITY OF FLORIDA
Fiscal Year: 2024
Award: $827,141
Funding agency: National Heart Lung and Blood Institute
Project Summary
Mesenchymal stem/stromal cells (MSCs) are under intensive investigation as therapeutics for treating a diverse
array of human diseases, and while some trials have demonstrated significant patient benefits, most have yielded
suboptimal outcomes. Currently, most clinical trials employ similar manufacturing schemes to treat medically
unrelated diseases, which reflects knowledge gaps in the critical quality attributes that confer potency and
undervalues the impacts of donor selection and manufacturing on potency. We previously identified TWIST1 as
a biomarker that quantifies inter-donor differences in stem/progenitor (growth, CFU-F activity, tri-lineage
potential) and paracrine (pro-angiogenic, anti-inflammatory, immuno-modulatory) functions of MSCs, thereby
linking these functions mechanistically. We then developed a Clinical Indications Prediction (CLIP) scale based
on TWIST1 that predicts inter-donor differences in MSC potency for a range of activities to produce MSC
products tailored to specific disease indications or patient populations. Most recently, we showed that TWIST1
levels correlate with donor stature, that MSCs from short statured donors exhibit poor growth and potent immuno-
modulatory activity, and that cGMP manufacturing platforms are biased against these donor populations. Based
on these findings, the central goals of this application are to further refine the CLIP scale and establish it as an
industry wide platform to inform donor selection and manufacturing processes to produce MSC-based
therapeutics of defined potency. To accomplish these goals, aim 1 will delineate epigenetic mechanisms that
confer inter-donor differences in TWIST1 levels and correlate outcomes with donor stature to establish stature
as a critical quality attribute that confers potency. MSCs from donors of varying stature will also be exposed to
agents that license pro-angiogenic (FGF2), anti-inflammatory (IL1, TNF) and immuno-modulatory (IFNG)
activities and their impacts on TWIST1 expression will be quantified to determine if the CLIP scale predicts their
sensitivity to these agents. These data will establish the importance of donor selection on MSC potency, which
to date has been largely overlooked. Aim 2 will subject MSCs from donors of variable stature to large-scale
expansion using standard (academic) and proprietary (industry) cGMP manufacturing platforms and quantify
impacts on potency pre- and post-manufacturing using well-established pre-clinical models. Methods (small
molecules, gene editing) to modulate TWIST1 levels post-manufacturing to alter potency will also be explored.
By revealing how biases in donor selection/manufacturing impact product potency these studies will highlight
limitations of current manufacturing platforms that reduce MSC therapeutic efficacy. Lastly, aim 3 will determine
if the CLIP scale predicts inter-donor differences in MSC-derived extracellular vesicle (EV) biogenesis, cargo
content, and potency. Expanding the CLIP scale to EVs will provide a platform to guide their manufacturing as
they enter clinical testing. Successful completion of these studies will deliver a robust, industry-wide platform to
produce MSC therapeutics of defined potency for treating specific disease indications.
Terms: <(TNF)-α><Acute Lung Injury><Acute Pulmonary Injury><Address><Adopted><Animal Model><Animal Models and Related Studies><Anti-Inflammatories><Anti-Inflammatory Agents><Anti-inflammatory><Assay><Basic Fibroblast Growth Factor><Basic Fibroblast Growth Factor Gene><Binding><Bioassay><Biogenesis><Biological Agent><Biological Assay><Biological Markers><Biological Products><Brittle Diabetes Mellitus><Cachectin><Cell Body><Cell Therapy><Cells><Clinical><Clinical Evaluation><Clinical Research><Clinical Study><Clinical Testing><Clinical Trials><Cyclic GMP><Data><Disease><Disorder><Donor Screening><Donor Selection><Epigenetic><Epigenetic Change><Epigenetic Mechanism><Epigenetic Process><Equation><Exhibits><Exposure to><FGF-2><FGF2><FGF2 gene><FGFB><Fibroblast Growth Factor 2><Fibroblast Growth Factor 2 Gene><Funding><Generalized Growth><Genes><Genomics><Goals><Growth><Guanosine Cyclic Monophosphate><HBGF-2><Heparin-Binding Growth Factor 2><Heparin-Binding Growth Factor Class II><Homolog of Drosophila TWIST 1><Homolog of Drosophila TWIST1><Human><IDDM><IFN-Gamma><IFN-g><IFN-γ><IFNG><IFNγ><IL-1><IL1><Immune Interferon><Immunomodulation><Industry><Industry Standard><Insulin-Dependent Diabetes Mellitus><Interferon Gamma><Interferon Type II><Interleukin I><Interleukin-1><Investigation><Juvenile-Onset Diabetes Mellitus><Ketosis-Prone Diabetes Mellitus><Knowledge><Laboratories><Licensing><Link><Lymphocyte-Stimulating Hormone><Macrophage Cell Factor><Macrophage-Derived TNF><Medical><Mesenchymal><Mesenchymal Progenitor Cell><Mesenchymal Stem Cells><Mesenchymal progenitor><Mesenchymal stromal/stem cells><Methods><Methylation><Modern Man><Molecular Interaction><Monitor><Monocyte-Derived TNF><Non-Polyadenylated RNA><Origin of Life><Outcome><Patient outcome><Patient-Centered Outcomes><Patient-Focused Outcomes><Patients><Population><Pre-Clinical Model><Preclinical Models><Process><Production><Prostate Epithelial Cell Growth Factor><RNA><RNA Gene Products><Ribonucleic Acid><Scheme><Stromal Cells><Sudden-Onset Diabetes Mellitus><T Helper Factor><T1 DM><T1 diabetes><T1D><T1DM><TNF><TNF A><TNF Alpha><TNF gene><TNF-stimulated gene 6 protein><TNF-α><TNFA><TNFα><TSG-6 protein><TWIST gene><TWIST1><TWIST1 gene><Testing><Therapeutic><Tissue Growth><Transcription Factor TWIST><Treatment Efficacy><Tumor Necrosis Factor><Tumor Necrosis Factor-alpha><Type 1 Diabetes Mellitus><Type 1 diabetes><Type I Diabetes Mellitus><bFGF><bio-markers><biologic marker><biologics><biomarker><biomarker identification><biopharmaceutical><biotherapeutic agent><cGMP><cell mediated therapies><cell-based therapeutic><cell-based therapy><cellular therapeutic><cellular therapy><clinical applicability><clinical application><clinical relevance><clinical test><clinically relevant><cohort><efficacy outcomes><epigenetically><extracellular vesicles><full scale manufacturing><histone modification><human disease><identification of biomarkers><identification of new biomarkers><immune modulation><immune regulation><immunologic reactivity control><immunomodulatory><immunoregulation><immunoregulatory><improved><improved outcome><insulin dependent diabetes><insulin dependent type 1><intervention efficacy><juvenile diabetes><juvenile diabetes mellitus><ketosis prone diabetes><lFN-Gamma><large scale manufacturing><large scale production><lymphocyte activating factor><manufacture><manufacturing process><marker identification><mass production><matrigel><mesenchymal stromal progenitor cells><mesenchymal-derived stem cells><model of animal><ontogeny><paracrine><patient oriented outcomes><patient population><patient stratification><population based><potency testing><pre-clinical><preclinical><progenitor><promoter><promotor><protein expression><research clinical testing><small molecule><stem><stratified patient><therapeutic efficacy><therapy efficacy><twist protein><type I diabetes><type one diabetes>