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Principal Investigator: Naoki Nakayama
Organization: STEADMAN PHILIPPON RESEARCH INSTITUTE
Fiscal Year: 2024
Award: $333,971
Funding agency: National Institute of Arthritis and Musculoskeletal and Skin Diseases
ABSTRACT
Impairment of articular cartilage function after injury and disease like osteoarthritis (OA), remains a major health
problem. One of the major drawbacks of tissue engineering-based therapies for damaged joint articular cartilage
is that the cartilaginous repair tissue formed by implanted mesenchymal stromal cells or endogenous progenitors
does not resemble articular cartilage, likely due to fibrochondrogenesis and the endochondral ossification
process. Joint cartilage is generated during embryogenesis by specialized GDF5+ cells called ‘interzone’ cells or
‘joint progenitors’. They are distinct from progenitors that give rise to growth plate chondrocytes. In this proposal,
we aim to define the molecular targets that control articular-like permanent chondrocyte formation versus growth
plate-like transient chondrocyte formation, by using novel GDF5+ mesenchymal cells developed from human
pluripotent stem cells (hPSCs). During in vitro chondrogenesis, such cells express signs of primitive (or
embryonic) articular chondrocytes but not of chondrocyte hypertrophy. Significantly, after transplantation of the
cartilage they develop, no mineralization was observed for 8 weeks (i.e., permanent cartilage). Therefore, the
hPSC-derived GDF5+ cells may share the activity of joint progenitors, although genome-wide RNA-sequencing
(seq) analyses suggested association with developing tenocytes or ligamentocytes. In contrast, alternative
hPSC-derived chondroprogenitors, SOX9+ cells, generated cartilage that readily underwent complete
mineralization, mimicking growth-plate chondroprogenitors. Interestingly, when mixed with GDF5+ cells, the
SOX9+ cell-derived cartilage behaved as permanent cartilage in a GDF5+ cell-dose-dependent manner,
suggesting the involvement of a non-cell autonomous mechanism. Therefore, we first propose to test if the
GDF5+ cells have a joint progenitor-like activity, characterize the (permanent) cartilage developed from them,
and shed lights on how the cells generate permanent cartilage (Aim 1). Second, we plan to identify genes
potentially involved in permanent cartilage formation from GDF5+ cells through comparative RNA-seq analyses
of cartilage pellets developed from the GDF5+ and SOX9+ cells (Aim 2). We will then functionally validate the
candidate genes (and their encoded proteins, inhibitors and activators if commercially available) for their ability
to enable SOX9+ cells to generate articular-like permanent chondrocytes using gene knockout and
overexpression techniques (Aim 3). We will then examine whether GDF5+ cells and such gene-modified SOX9+
cells induce more sustained repair of damaged joint cartilage than SOX9+ cells (Aim 4). Lastly, any of the genes
defined in these studies will be manipulated similarly in therapeutically relevant adult mesenchymal stromal cells
to confirm that targeting the same mechanisms will convert the adult stem cells to articular cartilage-forming cells
(Aim 4). Thus, success of the proposed research will provide mechanistic insights into how articular-like
permanent cartilage can be selectively formed from various chondrogenic cells, potentially leading to novel
therapeutic strategies for effective, sustained repair of damaged cartilage.
Terms: <21+ years old><3'5'-cyclic ester of AMP><Adenosine Cyclic 3',5'-Monophosphate><Adenosine Cyclic Monophosphate><Adenosine, cyclic 3',5'-(hydrogen phosphate)><Adult><Adult Human><Aging><Antibodies><Assay><Basal Transcription Factor><Basal transcription factor genes><Basic Research><Basic Science><Bio-Informatics><Bioassay><Bioinformatics><Biological Assay><Body Tissues><CRISPR approach><CRISPR based approach><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><Candidate Disease Gene><Candidate Gene><Cartilage><Cartilaginous Tissue><Cas nuclease technology><Cell Body><Cell Communication and Signaling><Cell Signaling><Cells><Characteristics><Chemicals><Chondrocytes><Chondrogenesis><Clustered Regularly Interspaced Short Palindromic Repeats approach><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><Coleonol><Cyclic AMP><Degenerative Arthritis><Degenerative polyarthritis><Disease><Disorder><Dose><Embryo><Embryo Development><Embryogenesis><Embryonic><Embryonic Development><Epiphyseal Plate><Epiphysial cartilage><Exhibits><Forskolin><Foundations><GDF5><GDF5 gene><Gene Modified><General Transcription Factor Gene><General Transcription Factors><Genes><Genomics><Growth Differentiation Factor 5 Gene><Growth Plate><Health><Hypercalcemic Hormone of Malignancy><Hypertrophy><Immunocompromised><Immunocompromised Host><Immunocompromised Patient><Immunosuppressed Host><Impairment><Implant><In Vitro><Injury><Intracellular Communication and Signaling><Joints><Knee><Mesenchymal><Methods><Mice><Mice Mammals><Molecular><Molecular Genetics><Molecular Target><Murine><Mus><Natural regeneration><Osteoarthritis><Osteoarthrosis><PTH Like Tumor Factor><PTH-Like Protein><PTH-Related Peptide><PTHrP><Paracrine Communication><Paracrine Signaling><Parathyroid Hormone Like Tumor Factor><Parathyroid Hormone-Like Hormone><Parathyroid Hormone-Like Protein><Parathyroid Hormone-Related Peptide><Patients><Physiologic Ossification><Physiological Ossification><Process><Production><Progenitor Cells><Property><Proteins><RNA Seq><RNA sequencing><RNAseq><Receptor Gene><Recombinant Parathyroid Hormone-Related Protein><Regeneration><Regulation><Regulator Genes><Research><Rodent Model><Sight><Signal Pathway><Signal Transduction><Signal Transduction Systems><Signaling><Site><Techniques><Technology><Testing><Therapeutic><Tissue Engineering><Tissues><Transcription Factor Proto-Oncogene><Transcription factor genes><Transcriptional Regulatory Elements><Transplantation><Tumor Hypercalcemic Factor><Validation><Vision><adenosine 3'5' monophosphate><adult progenitor><adult stem cell><adulthood><articular cartilage><bioengineered tissue><biological signal transduction><cAMP><candidate validation><cartilage development><cartilage graft><cartilage regeneration><cartilage repair><cartilage transplant><cartilage transplantation><cartilaginous><cell type><clinical relevance><clinically relevant><comparative><degenerative joint disease><engineered tissue><functional restoration><gene modification><genetically modified><genome scale><genome-wide><genomewide><human adult stem cell><human pluripotent stem cell><hypertrophic arthritis><immunosuppressed patient><in vivo><inhibitor><injured><injuries><insight><joint damage><joint injury><joint trauma><knockout gene><mesenchymal stromal cell><mineralization><new drug treatments><new drugs><new pharmacological therapeutic><new therapeutic approach><new therapeutic intervention><new therapeutic strategies><new therapeutics><new therapy><new therapy approaches><new treatment approach><new treatment strategy><next generation therapeutics><normal ossification><novel><novel drug treatments><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel therapeutic approach><novel therapeutic intervention><novel therapeutic strategies><novel therapeutics><novel therapy><novel therapy approach><ossification><osteoarthritic><overexpress><overexpression><paracrine><parathyroid hormone-related protein><physical property><progenitor><progenitor-like cell><regenerate><regeneration based therapy><regeneration therapy><regenerative therapeutics><regenerative therapy><regulatory gene><repair><repaired><restore function><restore functionality><restore lost function><small molecular inhibitor><small molecule inhibitor><somatic progenitor><somatic stem cell><stem cells><stem-like cell><success><therapeutically effective><tissue repair><trans acting element><transcription factor><transcriptome sequencing><transcriptomic sequencing><transcriptomics><transplant><validations><visual function>