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.
| Original language | English |
|---|---|
| Pages (from-to) | E695-E702 |
| Journal | American Journal of Physiology - Endocrinology and Metabolism |
| Volume | 307 |
| Issue number | 8 |
| DOIs | |
| State | Published - Oct 15 2014 |
Fingerprint
Keywords
- Aqueous
- CD34
- Progenitor cells
- Proliferative diabetic retinopathy
- Vitreous
ASJC Scopus subject areas
- Physiology
- Physiology (medical)
- Endocrinology, Diabetes and Metabolism
- Medicine(all)
Cite this
Growth factors/chemokines in diabetic vitreous and aqueous alter the function of bone marrow-derived progenitor (CD34+) cells in humans. / Balaiya, Sankarathi; Grant, Maria B.; Priluck, Joshua; Chalam, Kakarla V.
In: American Journal of Physiology - Endocrinology and Metabolism, Vol. 307, No. 8, 15.10.2014, p. E695-E702.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Growth factors/chemokines in diabetic vitreous and aqueous alter the function of bone marrow-derived progenitor (CD34+) cells in humans
AU - Balaiya, Sankarathi
AU - Grant, Maria B.
AU - Priluck, Joshua
AU - Chalam, Kakarla V.
PY - 2014/10/15
Y1 - 2014/10/15
N2 - 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.
AB - 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.
KW - Aqueous
KW - CD34
KW - Progenitor cells
KW - Proliferative diabetic retinopathy
KW - Vitreous
UR - http://www.scopus.com/inward/record.url?scp=84908043563&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84908043563&partnerID=8YFLogxK
U2 - 10.1152/ajpendo.00253.2014
DO - 10.1152/ajpendo.00253.2014
M3 - Article
C2 - 25159325
AN - SCOPUS:84908043563
VL - 307
SP - E695-E702
JO - American Journal of Physiology
JF - American Journal of Physiology
SN - 0193-1857
IS - 8
ER -