Highly Active Anti-Retroviral Therapy for HIV-infected individuals impedes bone mineral

Introduction: metabolism and increases prevalence of HAART-related bone disease

Infection with HIV-1, which develops into AIDs By Tara Manny
in almost all hosts, has been classified as a human Results:
pandemic for decades, eliciting life-threatening
• Osteoclastogenesis is associated with increased activities of
opportunistic infections to claim the lives of
millions. Advances in anti-retroviral therapies tartrate-resistant acid phosphatae (TRAP) promoter and NF-kB
(ART) have offered HIV-infected persons an transcription factor.
• Graph in Figure 1A displays how the osteoclastogenic effect of
opportunity for prolonged survival. However,
research has shown that extended exposure to AZT is concentration dependent, with less effect above and
protease inhibitors and nucleoside reverse below 5.0uL AZT +RANKL.
• Presence of AZT increases expression of TRAP and calcitonin
transcriptase inhibitors (NRTI) that compose
ARTs often lead to dangerous mitochondrial receptor (CTR), but RANK and GAPDH2 are unaffected
Figure 2: Osteoclastogenesis (FIG1B).
toxicities and the alteration of transcriptional
increased in PBMCs in presence of • Osteoclastogenesis increases most in the RANKL+AZT sample
processes. One prevalent side effect from highly
active anti-retroviral therapies (HAART) is the AZT. in HIV-positive HAART-naïve PBMCs (Fig2).
The effectiveness of AZT using human • Mice treated with AZT showed a reduction in BMD by 6.15 ±
pathogenesis of osteopenia and osteoporosis. Figure 4: Presence of AZT
peripheral blood mononuclear cells was 2.02% (Fig3A).
observed using HIV+ and HIV- patients. results in mitochondrial toxicity • Histomorphometric analysis shows that mice treated with AZT
but is reversed by application of had a significant increase in osteoclast surface and number.
PBMCs from 2 HIV-positive, HAART-naïve RANKL. • Figure 4A illustrates that RANKL treatment almost completely
patients and HIV-negative individuals were reverses mtDNA damages incurred by the presence of AZT.
(A) RAW264.7 cells were plated and
cultured with RANKL alone (R), or RANKL plus • Mitochondrial aconitase activity showed a 41%decrease in
treated with AZT and AZT plus
AZT (R+A) in the presence of M-CSF for 2 RAW264.7 cells treated with AZT from the control. Activity
RANKL for 1–3 days.
weeks. M-CSF alone treated PBMCs served as a was reversed or recovered by addition of RANKL, to
control (C). The number of multinucleated TRAP approximately 78% of the control(Fig4B).
• Mitochondrial DNA damage
positive osteoclasts were counted.
determined by qPCR. Frequency of
 Conclusion:
mtDNA damage induced by AZT and
AZT+RANKL expressed by
comparison to mtDNA damage in
• As levels of HAART increase and are made more readily available to
RANKL-treated cells. the HIV positive community, coincidences of osteopenia and
osteoporosis, among other complications, are likely to increase as
• (B) Aconitase activity of cell lysates well.
from RAW264.7 cells treated as
indicated for 3 days • Over-production of TRAP-positive osteoclasts leads to bone disease.
• (C) Western blot for mitochondrial
Figure 1: Osteoclastogenic effects by AZT SOD in RAW264.7 cells treated as • PBMC test results support the conclusion that HIV infection does not
indicated [for 1–3 days. itself accelerate osteoclastogenesis or the effect of AZT in vitro.
(Zidovudine) in RAW264.7 cells are
apparent. • AZT induces mtDNA damage while RANKL treatment completely
RAW264.7 cells were treated with AZT at various reverses mtDNA damage associated with AZT treatment.
concentrations in the presence of 50 ng/mL RANKL Figure 3: Presence of AZT decreases bone marrow density
for 3 days. TRAP staining assay gave a osteoclast • Since increased RANKL levels minimize the deleterious effects of
(BMD) and increases number and surface of osteoblasts.
measurement, as shown in the graph. Mice, 5 months old, were orally fed PBS (n = 6, control) or AZT (n 5, AZT 100 NRTIs on osteoclast mitochondria, it is possible that they promote
mg/kg/day) for 30 days. osteopenia/osteoporosis.
(B) For 1 to 3 days, RAW264.7 cells were treated • BMD measured by DEXA.
with RANKL (50 ng/mL, R) or RANKL plus AZT • Histomorphometry was analyzed in three vertebrae of each animal. Osteoclast • The osteoclastogenic effect of AZT requires the presence of RANKL.
(25 μM, R + A) and total mRNAs were isolated. parameters included number (OcN) and surface (OcS) of osteoclasts. Thus, interrupting RANKL by OPG, RANK-Fc, or other compounds
Reported as percentage change using AZT from control. may be potent therapeutic options.