Despite the large number of pregnant women and neonates infected with HIV receiving long-term treatment with antiretroviral agents, little is understood about the handling of these drugs in utero and in the neonatal kidney. Although the drugs are generally considered to be relatively safe for the fetus and neonate over the short-term, there is justifiable concern over undetected toxicities and long-term effects. This is part of a more general concern in pediatric pharmacology that unnecessary preterm and neonatal drug toxicity results from inappropriate dosing due to poorly understood mechanisms of maturation of drug handling capacity. With respect to drugs used for the treatment of HIV, in vitro data from the PI's group and others indicates that organic anion transporters (Oats), Oat1 and/or Oat3, are the key genes regulating antiretroviral handling. The PI's group was the first to identify Oat1, as well as a number of related genes, and has recently published the first adult Oat1 and Oat3 knockout mice data, which demonstrate defective organic anion (OA) transport and altered drug handling. However, the in vivo data in adult mice for certain OA compounds and drugs has revealed differences with in vitro data obtained mainly from Xenopus oocyte transport assays. Moreover, our recent developmental studies, using a novel ex-vivo method to analyze OA handling, suggest that Oat- independent mechanisms of OA transport may be important in utero and in neonates. Drawing on considerable preliminary data, in this REVISED application, we hypothesize that the mechanisms of in vivo OA drug transport are different in preterm and neonatal compared to adults. Furthermore, we hypothesize that, while Oat1 and Oat3 are the key antiretroviral transporters in adults and probably in the neonatal setting, in utero additional novel transporters also contribute to the handling of antiretrovirals used to treat HIV. These pathways need to be defined. We therefore aim to address the following questions: 1) What are the in vivo roles of Oat1 and Oat3 in net transport of antiretrovirals in the kidneys of neonatal wildtype and knockout animals? 2) Are additional pathways for transport of antiretrovirals active in utero? A combination of genetic, in vivo physiological, microarray and expression cloning methods will be employed to approach these questions, leveraging the joint strengths of the PI's group in OAT biology and kidney development. The results should provide key insights into the (likely different) mechanisms of drug handling in the in utero as well as neonatal settings and set the stage for translational studies. We have addressed all the criticisms of the prior application, including translational issues (please see letters from collaborators) and also provide new preliminary data. Despite the large number of pregnant women and neonates infected with HIV receiving long-term treatment with antiretroviral agents, little is understood about the handling of these drugs in utero and in the neonatal kidney. This application attempts to address the basic scientific aspects of antiretroviral drug handling in the immature kidney.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK079784-02
Application #
7568864
Study Section
AIDS Discovery and Development of Therapeutics Study Section (ADDT)
Program Officer
Kimmel, Paul
Project Start
2008-03-01
Project End
2012-02-29
Budget Start
2009-03-01
Budget End
2010-02-28
Support Year
2
Fiscal Year
2009
Total Cost
$328,313
Indirect Cost
Name
University of California San Diego
Department
Pediatrics
Type
Schools of Medicine
DUNS #
804355790
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Gallegos, Thomas F; Kouznetsova, Valentina; Kudlicka, Krystyna et al. (2012) A protein kinase A and Wnt-dependent network regulating an intermediate stage in epithelial tubulogenesis during kidney development. Dev Biol 364:11-21
Bush, Kevin T; Crawford, Brett E; Garner, Omai B et al. (2012) N-sulfation of heparan sulfate regulates early branching events in the developing mammary gland. J Biol Chem 287:42064-70
Vallon, Volker; Eraly, Satish A; Rao, Satish Ramachandra et al. (2012) A role for the organic anion transporter OAT3 in renal creatinine secretion in mice. Am J Physiol Renal Physiol 302:F1293-9
Gallegos, Thomas F; Martovetsky, Gleb; Kouznetsova, Valentina et al. (2012) Organic anion and cation SLC22 ""drug"" transporter (Oat1, Oat3, and Oct1) regulation during development and maturation of the kidney proximal tubule. PLoS One 7:e40796
Sweeney, Derina E; Vallon, Volker; Rieg, Timo et al. (2011) Functional maturation of drug transporters in the developing, neonatal, and postnatal kidney. Mol Pharmacol 80:147-54
Shah, Mita M; Sakurai, Hiroyuki; Gallegos, Thomas F et al. (2011) Growth factor-dependent branching of the ureteric bud is modulated by selective 6-O sulfation of heparan sulfate. Dev Biol 356:19-27
Garner, Omai B; Bush, Kevin T; Nigam, Kabir B et al. (2011) Stage-dependent regulation of mammary ductal branching by heparan sulfate and HGF-cMet signaling. Dev Biol 355:394-403
Torres, Adriana M; Dnyanmote, Ankur V; Bush, Kevin T et al. (2011) Deletion of multispecific organic anion transporter Oat1/Slc22a6 protects against mercury-induced kidney injury. J Biol Chem 286:26391-5
Wikoff, William R; Nagle, Megha A; Kouznetsova, Valentina L et al. (2011) Untargeted metabolomics identifies enterobiome metabolites and putative uremic toxins as substrates of organic anion transporter 1 (Oat1). J Proteome Res 10:2842-51
Nagle, Megha A; Truong, David M; Dnyanmote, Ankur V et al. (2011) Analysis of three-dimensional systems for developing and mature kidneys clarifies the role of OAT1 and OAT3 in antiviral handling. J Biol Chem 286:243-51

Showing the most recent 10 out of 20 publications