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2018 ; 497
(1
): 19-24
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Nephron segment-specific gene expression using AAV vectors
#MMPMID29407172
Asico LD
; Cuevas S
; Ma X
; Jose PA
; Armando I
; Konkalmatt PR
Biochem Biophys Res Commun
2018[Feb]; 497
(1
): 19-24
PMID29407172
show ga
AAV9 vector provides efficient gene transfer in all segments of the renal
nephron, with minimum expression in non-renal cells, when administered
retrogradely via the ureter. It is important to restrict the transgene expression
to the desired cell type within the kidney, so that the physiological endpoints
represent the function of the transgene expressed in that specific cell type
within kidney. We hypothesized that segment-specific gene expression within the
kidney can be accomplished using the highly efficient AAV9 vectors carrying the
promoters of genes that are expressed exclusively in the desired segment of the
nephron in combination with administration by retrograde infusion into the kidney
via the ureter. We constructed AAV vectors carrying eGFP under the control of:
kidney-specific cadherin (KSPC) gene promoter for expression in the entire
nephron; Na(+)/glucose co-transporter (SGLT2) gene promoter for expression in the
S1 and S2 segments of the proximal tubule; sodium, potassium, 2 chloride
co-transporter (NKCC2) gene promoter for expression in the thick ascending limb
of Henle's loop (TALH); E-cadherin (ECAD) gene promoter for expression in the
collecting duct (CD); and cytomegalovirus (CMV) early promoter that provides
expression in most of the mammalian cells, as control. We tested the specificity
of the promoter constructs in vitro for cell type-specific expression in mouse
kidney cells in primary culture, followed by retrograde infusion of the AAV
vectors via the ureter in the mouse. Our data show that AAV9 vector, in
combination with the segment-specific promoters administered by retrograde
infusion via the ureter, provides renal nephron segment-specific gene expression.