NAME: Rafael L. Ufret-Vincenty

eRA COMMONS USER NAME (credential, e.g., agency login): RUFRET

POSITION TITLE: Associate Professor in Ophthalmology, UT Southwestern Medical Center

EDUCATION/TRAINING

A.        Personal Statement

As a retina specialist and physician-scientist, I am interested in understanding the pathogenesis of retinal diseases.  I have had a long-standing interest in basic research and currently have 40% of my time protected for my basic research work.  My laboratory has generated a mouse model of early dry AMD based on the known association of the genetic variants in complement factor H.  We have also developed a simple new model of light induced retinal degeneration that is effective in mice expressing the 450 Met variant of RPE65.  We have shown that this model is mediated by oxidative stress and that it can serve as a model of retinal recovery from injury.  Currently we are using a forward genetics approach to search in an unbiased manner for novel genes important in retinal development and homeostasis and in retinal immune surveillance.  To this end we have established a collaboration with Dr. Bruce Beutler.  In this proposal, Dr. Beutler and I plan to apply a similar approach to discovering genes that modulate the development of early diabetic retinopathy.

  a. Chen B, Aredo B, Ding Yi, Zhong X, Zhu Y, Zhao CX, Kumar A, Xing C, Gautron L, Lyon S, Russell J, Li X, Tang M, Anderton P, Ludwig S, Moresco EMY, Beutler B, Ufret-Vincenty RL.  Forward genetic analysis using OCT screening identifies Sfxn3 mutations leading to progressive outer retinal degeneration in mice. PNAS In Print May 2020

  b. Chen B, Aredo B, Zhu Y, Ding Y, Xin-Zhao C, Ufret-Vincenty RL. A Mouse Model of Retinal Recovery From Photo-Oxidative/Photo-Inflammatory Injury: Nrf2, SOD1, DJ-1, and Parkin Are Not Essential to Recovery. Invest Ophthalmol Vis Sci. 2019;60:1165-1174.

  c. Ding Y, Aredo B, Zhong X, Zhao CX, Ufret-Vincenty RL. Increased susceptibility to fundus camera-delivered light-induced retinal degeneration in mice deficient in oxidative stress response proteins. Exp Eye Res. 2017 Mar 20;159:58-68.

B.        Positions and Honors

Positions

2016                                        Associate Professor of Ophthalmology - UT Southwestern Medical Center

2006                                        Assistant Professor of Ophthalmology - UT Southwestern Medical Center

Honors and Awards

2019-2025            NIH Study Section member, DPVS

2019-2022            UT Southwestern IRB member

2016,2017,2018   Listed in Texas Super Doctors, Texas Monthly magazine

2014-2016            David M. Crowley Foundation Award to Support AMD Research                      

2009-2012                     Disease Oriented Clinical Scholars Program          

1999-2001                     Student Director at the National Board of Directors- Alpha Omega Alpha

2000                                          Faculty Award - Most Outstanding Medical Student- UPR School of Medicine

2000                                          Summa Cum Laude – MD – UPR School of Medicine

2000                                          Dr. Ramon Ruiz Arnau Award for Clinical Research- UPR School of Medicine

C.        Contributions to Science
 

1. Forward genetics approach to understanding retinal physiology and disease:  Understanding of the mechanisms of retinal development and homeostasis is essential for the advancement of retinal regeneration strategies and conception of novel therapies for multifactorial retinal diseases.  Forward genetic analysis allows for the unbiased identification of non-redundant genes involved in the generation and maintenance of retinal integrity.  We are combining a special forward genetics protocol, in which all mice in tested pedigrees have been pre-genotyped at all mutant loci, with a sensitive and reproducible retinal screening assay, optical coherence tomography. This robust pipeline is allowing us to detect subtle phenotype-genotype associations which can then be characterized and explored mechanistically.

    a. Chen B, Aredo B, Ding Yi, Zhong X, Zhu Y, Zhao CX, Kumar A, Xing C, Gautron L, Lyon S, Russell J, Li X, Tang M, Anderton P, Ludwig S, Moresco EMY, Beutler B, Ufret-Vincenty RL.  Forward genetic analysis using OCT screening identifies Sfxn3 mutations leading to progressive outer retinal degeneration in mice. PNAS In Print May 2020

2. Models of age-related macular degeneration (AMD):  Despite important recent advances on the treatment of the neovascular form of AMD, clinical outcomes are still suboptimal.  Also, there is no effective therapy for geographic atrophy.  A better understanding of the mechanisms of disease is needed to develop new effective therapies.  My laboratory developed a mouse model of the early stages of the disease based on the complement factor H variant that increases the risk for the disease in humans.  Using this model we were able to show that expression of the “at risk” variant of CFH was sufficient to increase the sensitivity of the retina to oxidative stress and to lead to a pro-inflammatory phenotype of subretinal microglia.  I also participated on the conceptualization and early experimental design of another model of AMD based on immunization with an oxidation neo-epitope (Hollyfield et al. below).  Finally, I helped with the laser CNV experiments on a project that demonstrated that members of the microRNA-23~27~24 cluster are important in the process of choroidal neovascularization (Zhou et al.).

  a. Aredo B, Li T, Chen X, Zhang K, Xin-Zhao Wang C, Gou D, Zhao B, He Y and Ufret-Vincenty RL. A chimeric Cfh transgene leads to increased retinal oxidative stress, inflammation and accumulation of activated subretinal microglia in mice. Invest Ophthalmol Vis Sci. 2015;56:3427-40.

  b. Ufret-Vincenty RL, Aredo B, Liu X, McMahon A, Chen PW, Sun H, Niederkorn JY, Kedzierski W. Transgenic mice expressing variants of complement factor H develop AMD-like retinal findings. Invest Ophthalmol Vis Sci. 2010;51:5878-87. PubMed PMID: 20538999.

  c. Hollyfield JG, Bonilha VL, Rayborn ME, Yang X, Shadrach KG, Lu L, Ufret RL, Salomon RG, Perez VL. Oxidative damage-induced inflammation initiates age-related macular degeneration. Nat Med. 2008;14:194-8. PMCID: PMC2748836.

  d. Zhou Q, Gallagher R, Ufret-Vincenty R, Li X, Olson EN, Wang S. Regulation of angiogenesis and choroidal neovascularization by members of microRNA-23~27~24 clusters. Proc Natl Acad Sci USA. 2011;108:8287-92. PMCID: PMC3100947.

2. Understanding the role of microglia and RPE in retinal physiology and in the pathogenesis of AMD. RPE cells are key to retinal homeostasis, and are likely very important in the pathogenesis of AMD and other retinal diseases.  Most studies of gene expression in RPE cells involve either cell lines or RPE cell cultures.  We developed an easy and reproducible technique to generate RNA from RPE cells obtained from mouse eyes that leads to a high yield of highly pure RNA from RPE cells (no significant contamination from retina, choroid or sclera).

  a. Aredo B, Zhang K, Chen X, Wang CX, Li T, Ufret-Vincenty RL. Differences in the distribution, phenotype and gene expression of subretinal microglia/macrophages in C57BL/6N (Crb1(rd8/rd8)) versus C57BL6/J (Crb1(wt/wt)) mice. J Neuroinflammation. 2015;12:6. PMCID: PMC4305240.

  b. Xin-Zhao Wang C, Zhang K, Aredo B, Lu H, Ufret-Vincenty RL. Novel method for the rapid isolation of RPE cells specifically for RNA extraction and analysis. Exp Eye Res. 2012;102:1-9. PMCID: PMC3432720.

4. We are the first to describe that choroidal neovascular endothelium has exposure of phosphatidylserine (PS) in the outer leaflet of the cell membrane.  Furthermore, targeting exposed PS with antibodies can lead to reduction in choroidal angiogenesis as seen in the laser CNV model and the ex vivo choroidal sprouting assay. These findings open the possibility of combination therapies that could increase the treatment efficacy and decrease the treatment load on patients with AMD.

a. Li T, Aredo B, Zhang K, Zhong X, Pulido JS, Wang S, He YG, Huang X, Brekken RA, Ufret-Vincenty RL. Phosphatidylserine (PS) Is Exposed in Choroidal Neovascular Endothelium: PS-Targeting Antibodies Inhibit Choroidal Angiogenesis In Vivo and Ex Vivo. Invest Ophthalmol Vis Sci. 2015;56:7137-45. PMCID: PMC4634629.

5. Inflammatory responses directed against microorganisms may lead to injury to self-tissues via the phenomenon of molecular mimicry.  Using the model of experimental autoimmune encephalomyelitis I was able to show that T cells directed against various viral peptides could cross-react with myelin basic protein and lead to CNS inflammation and MS-like disease.  Furthermore, these cells could survive in vivo for months and be reactivated by exposure to the same or other cross-reacting viral peptides and lead to disease.  These observations are relevant to MS, but also were important in our generation of the CEP-adduct immunization model of early AMD.

a. Ufret-Vincenty RL, Quigley L, Tresser N, Pak S, Gado A, Hausmann S, Wucherpfennig KW, Brocke S. In vivo survival of viral antigen-specific T cells that induce experimental autoimmune encephalomyelitis. J Exp Med. 1998;188:1725-38. PMCID: PMC2212527.

Complete List of Published Work in MyBibliography:   https://www.ncbi.nlm.nih.gov/sites/myncbi/1nQ3gs-PzbU5g/bibliography/47992531/public/?sort=date&direction=descending

D.        Additional Information: Research Support and/or Scholastic Performance

Patricia and Col. William Massad Retina Research Fund    Ufret-Vincenty (PI)                   2018-2021

The goal of this grant is to support research to understand retinal degeneration

R01EY022652                Ufret-Vincenty (PI)                                                          9/1/2012-8/31/2017           

Role of complement factor H and immunity in AMD: a novel transgenic model.

-Goals: 1. study the role of a complement factor H (CFH) variant in determining the susceptibility of the retina/RPE to oxidative stress and inflammation, 2. understand the role of macrophages/microglia in disease, 3. determine the relevance of molecular interactions between CFH and Crp, malondialdehyde and glycosaminoglycans in the development of disease.

David M. Crowley Foundation AMD Research Fund     Ufret-Vincenty (PI)                          2013-2015 

The goal of this grant is support the development of animal models of AMD.