HIV integrase inhibitor, Elvitegravir, impairs RAG functions and inhibits V(D)J recombination

Abstract

Integrase inhibitors are a class of antiretroviral drugs used for the treatment of AIDS that target HIV integrase, an enzyme responsible for integration of viral cDNA into host genome. RAG1, a critical enzyme involved in V(D)J recombination exhibits structural similarity to HIV integrase. We find that two integrase inhibitors, Raltegravir and Elvitegravir, interfered with the physiological functions of RAGs such as binding, cleavage and hairpin formation at the recombination signal sequence (RSS), though the effect of Raltegravir was limited. Circular dichroism studies demonstrated a distinct change in the secondary structure of RAG1 central domain (RAG1 shares DDE motif amino acids with integrases), and when incubated with Elvitegravir, an equilibrium dissociation constant (Kd) of 32.53±2.9 μM was determined by Biolayer interferometry, leading to inhibition of its binding to DNA. Besides, using extrachromosomal assays, we show that Elvitegravir inhibited both coding and signal joint formation in pre-B cells. Importantly, treatment with Elvitegravir resulted in significant reduction of mature B lymphocytes in 70% of mice studied. Thus, our study suggests a potential risk associated with the use of Elvitegravir as an antiretroviral drug, considering the evolutionary and structural similarities between HIV integrase and RAGs.

Biochemical activity of RAGs is impeded by Dolutegravir, an HIV integrase inhibitor

“…A recent report has shown that Elvitegravir, an integrase inhibitor, can inhibit RAG functions both in vitro and in vivo, but not Raltegravir 20 . In the present study, we have investigated the impact of second-generation integrase inhibitor, Dolutegravir, on V(D)J recombinase and its action.…”

“…Mice fed with Elvitegravir showed a decrease in maturation of B cells 20 . A significant abrogation of RAG1 activity in vitro and in vivo was observed for Elvitegravir, on the other hand Raltegravir caused limited abrogation of RAG1 activity in vitro 20 . There are also reports of high rate of Non-Hodgkin's lymphoma in patients undergoing treatment with integrase inhibitors as well as early onset of autoimmune diseases 21,22 .…”

“…Both these compounds act by binding to the integrase-viral DNA complex, thus inhibiting the strand transfer reaction but not the 3′-processing reaction 19 . Elvitegravir and Raltegravir, were studied previously and it was shown that Elvitegravir impeded RAG activity in vitro as well as in vivo 20 . Mice fed with Elvitegravir showed a decrease in maturation of B cells 20 .…”

“…Elvitegravir and Raltegravir, were studied previously and it was shown that Elvitegravir impeded RAG activity in vitro as well as in vivo 20 . Mice fed with Elvitegravir showed a decrease in maturation of B cells 20 . A significant abrogation of RAG1 activity in vitro and in vivo was observed for Elvitegravir, on the other hand Raltegravir caused limited abrogation of RAG1 activity in vitro 20 .…”

“…Based on the previous reports of effect of integrase inhibitors on RAG1 functions and in turn V(D)J recombination 5,20 , in the current study, we investigated the effects of Dolutegravir using biochemical assays. Here we report that unlike Elvitegravir, Dolutegravir has several fold lower effect on RAG functions.…”