title: Growth factors/chemokines in diabetic vitreous and aqueous alter the function of bone marrow-derived progenitor (CD34⁺) cells in humans.
abstract: Ocular ischemic microenvironment plays a critical role in the progression of diabetic retinopathy (DR). In this study, we investigated the effect of vitreous and aqueous obtained from proliferative DR patients on the function of CD34⁺ cells derived from healthy humans. Human CD34⁺ cells were incubated with vitreous or aqueous of subjects with PDR. After incubation, cell migration of CD34⁺ was evaluated with CXCL12. Intracellular levels of nitric oxide (NO) were measured with DAF-FM. Tube formation assay was used to evaluate the effect of treated CD34⁺ cells on in vitro angiogenesis. Angiogenic protein array and mass spectrometry (MS) were performed to ascertain the factors secreted by healthy nondiabetic CD34⁺ cells exposed to diabetic vitreous or aqueous. PDR vitreous/aqueous reduced migration of CD34⁺ cells (672.45 ± 42.1/736.75 ± 101.7 AFU; P < 0.01) and attenuated intracellular NO levels (182 ± 1.4/184.5 ± 6.3 AFU, P = 0.002). Pretreatment with PDR vitreous suppressed tube formation of human retinal endothelial cells (64 ± 1.6 vs. 80 ± 2.5). CD34⁺ exposed to PDR vitreous resulted in the increased expression of CXCL4 and serpin F1, whereas CD34⁺ exposed to PDR aqueous showed increased expression of CXCL4, serpin F1, and endothelin-1 (ET-1). MS analysis of CD34⁺ (exposed to PDR vitreous) expressed J56 gene segment, isoform 2 of SPARC-related modular calcium-binding protein 2, isoform 1 of uncharacterized protein c1 orf167, integrin α-M, and 40s ribosomal protein s21. Exposure of healthy nondiabetic CD34⁺ cells to PDR vitreous and aqueous resulted in decreased migration, reduced generation of NO, and altered paracrine secretory function. Our results suggest that the contribution of CD34⁺ cells to the aberrant neovascularization observed in PDR is driven more by the proangiogenic effects of the retinal cells rather than the influence of the vitreous.
authors.affiliation: Department of Ophthalmology, University of Florida College of Medicine, Jacksonville, Florida; and.
authors.forename: Sankarathi
authors.initials: S
authors.lastname: Balaiya
authors.affiliation: Eugene and Marilyn Glick Eye Institute, Indiana University, Indianapolis, Indiana.
authors.forename: Maria B
authors.initials: MB
authors.lastname: Grant
authors.affiliation: Department of Ophthalmology, University of Florida College of Medicine, Jacksonville, Florida; and.
authors.forename: Joshua
authors.initials: J
authors.lastname: Priluck
authors.affiliation: Department of Ophthalmology, University of Florida College of Medicine, Jacksonville, Florida; and kchalam@jax.ufl.edu.
authors.forename: Kakarla V
authors.initials: KV
authors.lastname: Chalam
mesh_terms: D000328:Adult; D053687:Adult Stem Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D000368:Aged; D018952:Antigens, CD34 / Q000378:metabolism; D001082:Aqueous Humor / Q000276:immunology / Q000378:metabolism*; D001854:Bone Marrow Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D002478:Cells, Cultured; D002633:Chemotaxis; D003930:Diabetic Retinopathy / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D019332:Endothelin-1 / Q000819:agonists / Q000737:chemistry / Q000378:metabolism*; D005136:Eye Proteins / Q000819:agonists / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008875:Middle Aged; D009389:Neovascularization, Pathologic / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D009414:Nerve Growth Factors / Q000819:agonists / Q000737:chemistry / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism; D010449:Peptide Mapping; D010978:Platelet Factor 4 / Q000819:agonists / Q000737:chemistry / Q000378:metabolism*; D012160:Retina / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D015843:Serpins / Q000819:agonists / Q000737:chemistry / Q000378:metabolism*; D018630:Vitreoretinopathy, Proliferative / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D014822:Vitreous Body / Q000276:immunology / Q000378:metabolism*; D000099174:Pigment Epithelium-Derived Factor
chemical_list: D018952:Antigens, CD34; D019332:Endothelin-1; D005136:Eye Proteins; D009414:Nerve Growth Factors; D009569:Nitric Oxide; D010978:Platelet Factor 4; D015843:Serpins; D000099174:Pigment Epithelium-Derived Factor; C064936:PF4V1 protein, human
keywords: CD34+; aqueous; progenitor cells; proliferative diabetic retinopathy; vitreous