Stem cell therapy finds an ideal proving ground in the eye, an organ with relative immune privilege that is accessible yet isolated. Therapies for the eye are generally neither invasive nor systemic; and since the eye is optically transparent, treatment can be monitored easily and non-invasively in living animals-a major advantage over other systems. In preclinical studies of retinal disease, grafts of healthy retinal pigment epithela (RPE) can restore damaged retina. Looking ahead to clinical applications of RPE grafting, we posit that RPE grafts grown from autologous stem cells (a patient's own stem cells) would be the optimal approach to develop. Nevertheless, clinical use of this method awaits resolution of several knowledge gaps: Are an adult's stem cells pluripotent enough to render functional RPE grafts? Even if patient grafts are functional, will in vitro culture make them antigenic? Also, eve if autologous RPE grafts work, will gene therapy be more effective at late stages of RP? These questions are addressed by Aims 1, 2, and 3, respectively. The therapies explored here aim to repair hereditary retinal degeneration. Although, in general our results will pertain to retinitis pigmentosa (RP), these studies focus on RP caused by rare mutations in membrane frizzled- related gene (MFRP). MFRP retinopathy is ideal for our studies for three reasons. First, known MFRP point mutations cause an RP phenotype. In addition, MFRP retinopathy has a decades-long window of opportunity for treatment: Over the life span, electroretinogram (ERG) changes lag behind photoreceptor loss, suggesting that despite retinal damage, MFRP retinopathy patients retain sight and are likely treatable until late stages of the disease. The third reason fr choosing MFRP retinopathy is that we can take advantage of the well characterized, MFRP mutant mouse line, rd6 that also suffers blinding RP. For our study, MFRP-deficient patient stem cells will be isolated, cultured, and transduced with an Adeno-Associated-virus (AAV) to express wild-type MFRP. These MFRP gene repaired patient stem cells will be differentiated into RPE and grafted into right eyes of immunodeficient (Scid), MFRP-deficient (rd6) mice. Left and right eyes will be compared for reversal of the RP phenotype. Our long-term goals are to create cell- and gene-therapy cures for hereditary retinal diseases, and develop strategies that extend to other diseases. Our objective in this proposal is to find ways to use patient stem cells as a source of retinal grafts. We will test our central hypothesis that patient- derived RPE grafts, repaired by AAV in vitro, are already a therapeutic option.

Public Health Relevance

The proposed research is relevant to public health because the goal is to develop gene therapy approaches that explore in vivo gene editing and correction tools for the eye to treat retinitis pigmentosa, which are inherited eye disorders that ultimately cause blindness. Thus, the proposed research is relevant to NEI's 'Audacious Goals' pertaining to research with respect to blinding eye diseases and preservation of sight.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY026682-02
Application #
9244782
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Neuhold, Lisa
Project Start
2016-04-01
Project End
2020-03-31
Budget Start
2017-04-01
Budget End
2018-03-31
Support Year
2
Fiscal Year
2017
Total Cost
Indirect Cost
Name
University of Iowa
Department
Pediatrics
Type
Schools of Medicine
DUNS #
062761671
City
Iowa City
State
IA
Country
United States
Zip Code
52242
Jauregui, Ruben; Park, Karen Sophia; Tsang, Stephen H (2018) Two-year progression analysis of RPE65 autosomal dominant retinitis pigmentosa. Ophthalmic Genet 39:544-549
Velez, Gabriel; Tang, Peter H; Cabral, Thiago et al. (2018) Personalized Proteomics for Precision Health: Identifying Biomarkers of Vitreoretinal Disease. Transl Vis Sci Technol 7:12
Jauregui, Ruben; Park, Karen Sophia; Duong, Jimmy K et al. (2018) Quantitative progression of retinitis pigmentosa by optical coherence tomography angiography. Sci Rep 8:13130
Liu, Katherine Y; Sengillo, Jesse D; Velez, Gabriel et al. (2018) Missense mutation in SLIT2 associated with congenital myopia, anisometropia, connective tissue abnormalities, and obesity. Orphanet J Rare Dis 13:138
DiCarlo, James E; Mahajan, Vinit B; Tsang, Stephen H (2018) Gene therapy and genome surgery in the retina. J Clin Invest 128:2177-2188
Cui, Xuan; Jauregui, Ruben; Park, Karen Sophia et al. (2018) Multimodal characterization of a novel mutation causing vitamin B6-responsive gyrate atrophy. Ophthalmic Genet 39:512-516
Sengillo, Jesse D; Lee, Winston; Bilancia, Colleen G et al. (2018) Phenotypic expansion and progression of SPATA7-associated retinitis pigmentosa. Doc Ophthalmol 136:125-133
Jauregui, Ruben; Thomas, Amanda L; Liechty, Benjamin et al. (2018) SCAPER-associated nonsyndromic autosomal recessive retinitis pigmentosa. Am J Med Genet A :
Cho, Galaxy Y; Schaefer, Kellie A; Bassuk, Alexander G et al. (2018) CRISPR GENOME SURGERY IN THE RETINA IN LIGHT OF OFF-TARGETING. Retina 38:1443-1455
Petersen-Jones, Simon M; Occelli, Laurence M; Winkler, Paige A et al. (2018) Patients and animal models of CNG?1-deficient retinitis pigmentosa support gene augmentation approach. J Clin Invest 128:190-206

Showing the most recent 10 out of 54 publications