Document text
Principal Investigator: KARL J WAHLIN
Organization: UNIVERSITY OF CALIFORNIA, SAN DIEGO
Fiscal Year: 2024
Award: $395,000
Funding agency: National Eye Institute
SUMMARY
Retinal degenerative (RD) diseases, such as Retinitis pigmentosa (RP) and Leber congenital amaurosis
(LCA), cause dysfunction and cell death of photoreceptor (PR) cells, ultimately leading to blindness. LCA
is the leading cause of inherited childhood blindness resulting in a loss of vision at or soon after birth. Though
this is considered to be quite rare, these blinding diseases are devastating for those affected. Current efforts are
being made to develop gene-therapies aimed at correcting some of the genes affected in RD and this approach
has shown some promise in animals and humans for restoring RPE65 gene expression, but there are many
other causes of RD for which there is no cure. In addition, due to the many mutations involved in RD, there are
significant gaps in our understanding of how PR loss occurs. To address this, we will use human pluripotent
stem cell (PSC) based retinal cell-reporter lines with RD-associated alleles to help explore the mechanisms of
PR cell death. Given the typically long period of time required to generate human retinas in the laboratory, the
severity and rapid onset of degeneration in LCA makes it an attractive experimental model to study human RD
and to develop potential therapies. We will study the aryl hydrocarbon receptor interacting protein-like1 (AIPL1)
gene to explore three functional domains that harbor naturally occurring mutations in patients with LCA and
cone-rod dystrophy (CORD). A comparative analysis of different mutations might lead to a better understanding
of how rods and cones die and greater insight into other more common forms of PR degeneration, such as age-
related macular degeneration (AMD). A central hypothesis is that human PSC derived 3D retina organoids
with AIPL1 mutations will recapitulate human retinal dystrophy resulting in PR loss. This hypothesis is
supported by our recent work, and others, showing that human PSCs can be coaxed into becoming retinal
eyecup-like structures with PRs, a laminar morphology and outer segment structures that are similar to an actual
retina. This proposal will bridge two innovative technologies; (1) genome-editing to generate genetically
matched retinal reporter PSC derived retinas with disease-associated mutations and (2) gene-correction to
repair genetic defects and promote PR cell survival. Given the very early onset of LCA it is important to define
the appropriate windows of time for such treatment options. Not only will these studies lead to new insights
into the biology of RD disease, but could also provide an innovative resource to develop therapies for
the treatment of RD.
Terms: <3-D><3-Dimensional><3D><ARA9 protein><Address><Affect><Age related macular degeneration><Age-Related Maculopathy><Alleles><Allelomorphs><Animals><Apoptosis><Apoptosis Pathway><Binding><Binding Proteins><Biologic Models><Biological Models><Biology><Birth><Blindness><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><Cas nuclease technology><Cell Body><Cell Cycle Proteins><Cell Death><Cell Division Cycle Proteins><Cell Survival><Cell Viability><Cell-Cycle Regulatory Proteins><Cells><Childhood><Clinical><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><Complex><Cone><Cyclic GMP><DNA Damage Repair><DNA Repair><DNA Therapy><Data><Defect><Disease><Disorder><Dysfunction><Electrophysiology><Electrophysiology (science)><Experimental Models><Eye diseases><Fluorescence><Freezing><Functional disorder><Gene Expression><Gene Transfer Clinical><Genes><Genetic Alteration><Genetic Change><Genetic Intervention><Genetic defect><Goals><Guanosine Cyclic Monophosphate><Hereditary><Human><Immunoblotting><Inherited><Knock-out><Knockout><Knowledge><Label><Laboratories><Lead><Leber congenital amaurosis><Leber's amaurosis><Leber's congenital amaurosis><Ligand Binding Protein><Ligand Binding Protein Gene><Measures><Mice><Mice Mammals><Model System><Modeling><Modern Man><Molecular><Molecular Interaction><Monitor><Morphology><Murine><Mus><Mutation><Neurophysiology / Electrophysiology><Organoids><Parturition><Pathogenesis><Pathway interactions><Patients><Pb element><Phenotype><Photoreceptor Cell><Photoreceptors><Photosensitive Cell><Physiopathology><Pigmentary Retinopathy><Pluripotent Stem Cells><Population><Primates><Primates Mammals><Progenitor Cells><Programmed Cell Death><Property><Protein Binding><Proteins><RPE65><RPE65 protein><Recombinant adeno-associated virus><Recombinant adeno-associated virus (rAAV)><Recovery of Function><Reporter><Research Resources><Resources><Retina><Retinal Degeneration><Retinal Diseases><Retinal Disorder><Retinal Dystrophy><Retinitis Pigmentosa><Rod><Rods and Cones><Role><Severities><Sorting><Structure><Study models><System><Tapetoretinal Degeneration><Testing><Time><Tissues><Unscheduled DNA Synthesis><Variant><Variation><Vertebrate Photoreceptors><Viral><Visual Receptor><Western Blotting><Western Immunoblotting><Work><XAP2 protein><age dependent macular degeneration><age induced macular degeneration><age related macular disease><age related macular dystrophy><amaurosis congenita of Leber><aryl hydrocarbon receptor-interacting protein><bound protein><cGMP><cdc Proteins><cell type><comparative><cone-rod dystrophy><congenital amaurosis of retinal origin><degenerative retina diseases><develop therapy><disease phenotype><disease-in-a-dish><early onset><electrophysiological><eye disorder><functional recovery><gene corrected><gene correction><gene editing method><gene editing methodology><gene editing strategy><gene editing techniques><gene repair therapy><gene therapy><gene-based therapy><gene-editing approach><genetic therapy><genome editing><genome mutation><genomic correction><genomic editing><genomic therapy><heavy metal Pb><heavy metal lead><hepatitis B virus-associated protein 2><human disease><human pluripotent stem cell><human progenitor cell derived><human stem cell-derived><in vivo><inherited retinal degeneration><innovate><innovation><innovative><innovative technologies><insight><intervention development><knockout gene><live cell image><live cell imaging><live cellular image><live cellular imaging><mRNA Expression><mutant><necrocytosis><ocular disease><ocular disorder><ophthalmopathy><pathophysiology><pathway><pediatric><photoreceptor degeneration><pluripotent progenitor><polyproline><prevent><preventing><protein blotting><rAAV><recombinant AAV><repair><repaired><response><retina degeneration><retina disease><retina disorder><retinal degenerative><retinal degenerative diseases><retinopathy><rod and cone dystrophy><rod-cone dystrophy><selective expression><selectively expressed><senile macular disease><social role><stem cell derived tissues><stem cells><therapy development><three dimensional><treatment development><vision loss><visual loss>