The candidate is applying for a Mentored Clinical Scientist Research Career Development Award (K08) to support a 5 year research career training program that will significantly bolster his transition to faculty independence as a fellowship trained clinician-scientist specializing in vitreoretinal and macular diseases. The rigorous training program has been designed with extensive input and unyielding support from the candidate's mentor, co-mentor, chairman, and advisory committee. All training will take place in the excellent scientific and academic environment of the University of Kentucky, Department of Ophthalmology and Visual Sciences which includes an ocular angiogenesis research consortium consisting of four independent investigators. The applicant will spend the first 3 years of the program as a Research Assistant Professor and Clinical Fellow. The specific design of the fellowship is precisely detailed in the Career Development section and accompanied by a very strong institutional commitment that allows the candidate at least 80% effort performing research and 20% performing clinical duties. After completion of the fellowship, the candidate will transition to a full-time faculty position as Assistant Professor at University of Kentucky with the same institutional commitment of 80% research and 20% clinical effort. The long-term goal of the award is to offer the candidate the critical career development support to establish a pathway to faculty independence that will extensively train him as a biomedical researcher, confident and masterful specialist in the diagnosis and treatment of vitreoretinal and macular diseases, and leading academician. The candidate aims to have an independent laboratory up and running within the initial 12-18 months of the award period in order to gain early independence with his research project and develop critical principal investigator skills with guidance from his established mentors and advisory committee. The research plan has been devised by the candidate and his mentors and aims to extend his scientific expertise beyond basic vascular biology with a rich experience to develop new techniques and to utilize such methods in experimental designs to obtain critical data that expand his current fund of knowledge in order to yield fundamental insights on endothelial cell dysregulation, immuno-modulation in vascular growth, and therapeutic targets in neovascular age-related macular degeneration (AMD). AMD is a devastating blinding disease that affects as many as 10 million Americans, a number that rivals the prevalence of all cancers combined and is unfortunately expected to double by 2020. The overwhelming cause of severe vision loss in AMD is choroidal neovascularization (CNV), the growth of abnormal blood vessels beneath the retina. Despite the use of recently approved molecular therapeutics targeting vascular endothelial growth factor (VEGF)-A, the majority of patients do not regain functional vision, and a significant fraction progress to legal blindness. This treatment failure is attributed to the frequent clinical situation in which patients present to eye physicians after vision is already lost and irreversible structural and functional tissue damage has already occurred. Moreover, while patients with AMD often undergo at least one or more dilated eye examinations per year, the physician is often not able to detect CNV before it grows beneath the retina and wreaks havoc. Fortunately, there is an exciting development that may provide the first available technique for detecting incipient CNV in patients with AMD thereby allowing for earlier detection and treatment to prevent loss of vision and quality of life. This discovery was recently reported in the journal Nature and described as the first molecular marker that is specific for human CNV (Takeda et al. Nature 2009). It is an immune receptor called chemokine receptor 3 (CCR3) that is involved in allergic response and recruitment of white blood cells but may also be found on inflamed and growing blood vessels. The identification of CCR3 as a molecular signature has opened a gateway to a new understanding in vascular disease in AMD. CCR3 expression is found on the surface of the specialized endothelial cells that line the abnormal blood vessels. In this proposal, rigorous basic science investigations of the cellular circuitry that mediates this pathogenic switch will be explored in conjunction with the biological relationships between endothelial cell dysregulation, aging, immune activation, and oxidative stress. The initial aim of the study is to decipher how these established AMD risk factors regulate the progression of disease through chemokine and other immune signaling pathways in order to develop appropriately targeted diagnostics and therapeutics that will enable clinicians to offer improved care of patients suffering from AMD.
A second aim focuses specifically on CCR3 biology with scientific exploration of its pathogenetic induction, expression patterns and signaling effects during endothelial cell dysregulation in neovascular AMD. In a final aim, optimized fluorescent CCR3- targeting probes will be engineered and applied to pre-clinical animal models of CNV and human cellular toxicology studies in order to obtain necessary data to proceed with future clinical studies. These specific experimental aims will provide fundamental insights into the modulatory function of the immune system, chemokine signaling, and the role of CCR3 as a molecular switch during the endothelial cell dysregulation that heralds vision loss in neovascular AMD.

Public Health Relevance

The abnormal growth of blood vessels beneath the retina results in devastating vision loss in age-related macular degeneration (AMD), yet the specific molecular and cellular mechanisms by which this process occurs are an area of active scientific investigation. The proposal aims to provide fundamental insights into the dysregulation of the specialized endothelial cells that line the abnormal blood vessels during AMD progression. Importantly, this award will grant the candidate a critical opportunity to gain essential research training while greatly enhancing career development and a confident transition to faculty independence.

National Institute of Health (NIH)
National Eye Institute (NEI)
Clinical Investigator Award (CIA) (K08)
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Study Section
Special Emphasis Panel (ZEY1)
Program Officer
Agarwal, Neeraj
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University of Kentucky
Schools of Medicine
United States
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Kleinman, Mark E; Ambati, Jayakrishna (2016) Complement Activation and Inhibition in Retinal Diseases. Dev Ophthalmol 55:46-56
Ouseph, Madhu M; Kleinman, Mark E; Wang, Qing Jun (2016) Vision loss in juvenile neuronal ceroid lipofuscinosis (CLN3 disease). Ann N Y Acad Sci 1371:55-67
Berner, Andre K; Kleinman, Mark E (2016) Therapeutic Approaches to Histone Reprogramming in Retinal Degeneration. Adv Exp Med Biol 854:39-44
Gelfand, Bradley D; Wright, Charles B; Kim, Younghee et al. (2015) Iron Toxicity in the Retina Requires Alu RNA and the NLRP3 Inflammasome. Cell Rep 11:1686-93
Frederick, Paul A; Kleinman, Mark E (2014) The Immune System and AMD. Curr Ophthalmol Rep 2:14-19
Kleinman, Mark E; Ambati, Jayakrishna (2013) Clocking in on diabetic retinopathy. Diabetes 62:29-30
Dridi, Sami; Hirano, Yoshio; Tarallo, Valeria et al. (2012) ERK1/2 activation is a therapeutic target in age-related macular degeneration. Proc Natl Acad Sci U S A 109:13781-6
Kleinman, Mark E; Kaneko, Hiroki; Cho, Won Gil et al. (2012) Short-interfering RNAs induce retinal degeneration via TLR3 and IRF3. Mol Ther 20:101-8
Tarallo, Valeria; Hirano, Yoshio; Gelfand, Bradley D et al. (2012) DICER1 loss and Alu RNA induce age-related macular degeneration via the NLRP3 inflammasome and MyD88. Cell 149:847-59
Kleinman, Mark E; Baffi, Judit Z; Ambati, Jayakrishna (2010) The multifactorial nature of retinal vascular disease. Ophthalmologica 224 Suppl 1:16-24