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Human Exportin-1 is a Target for Combined Therapy of HIV and AIDS Related Lymphoma.

Boons E, Vanstreels E, Jacquemyn M, Nogueira TC, Neggers JE, Vercruysse T, van den Oord J, Tamir S, Shacham S, Landesman Y, Snoeck R, Pannecouque C, Andrei G, Daelemans D - EBioMedicine (2015)

Bottom Line: Here we report on the dual anti-HIV and anti-PEL effect of targeting a single process common in both diseases.At the same time, SINE caused the nuclear accumulation of p53 tumor suppressor protein as well as inhibition of NF-κB activity in PEL cells resulting in cell cycle arrest and effective apoptosis induction.Our findings provide strong rationale for inhibiting XPO1 as an innovative strategy for the combined anti-retroviral and anti-neoplastic treatment of HIV and PEL and offer perspectives for the treatment of other AIDS-associated cancers and potentially other virus-related malignancies.

View Article: PubMed Central - PubMed

Affiliation: KU Leuven, Department of Microbiology and Immunology, Laboratory of Virology and Chemotherapy, Rega Institute for Medical Research, B-3000 Leuven, Belgium.

ABSTRACT
Infection with HIV ultimately leads to advanced immunodeficiency resulting in an increased incidence of cancer. For example primary effusion lymphoma (PEL) is an aggressive non-Hodgkin lymphoma with very poor prognosis that typically affects HIV infected individuals in advanced stages of immunodeficiency. Here we report on the dual anti-HIV and anti-PEL effect of targeting a single process common in both diseases. Inhibition of the exportin-1 (XPO1) mediated nuclear transport by clinical stage orally bioavailable small molecule inhibitors (SINE) prevented the nuclear export of the late intron-containing HIV RNA species and consequently potently suppressed viral replication. In contrast, in CRISPR-Cas9 genome edited cells expressing mutant C528S XPO1, viral replication was unaffected upon treatment, clearly demonstrating the anti-XPO1 mechanism of action. At the same time, SINE caused the nuclear accumulation of p53 tumor suppressor protein as well as inhibition of NF-κB activity in PEL cells resulting in cell cycle arrest and effective apoptosis induction. In vivo, oral administration arrested PEL tumor growth in engrafted mice. Our findings provide strong rationale for inhibiting XPO1 as an innovative strategy for the combined anti-retroviral and anti-neoplastic treatment of HIV and PEL and offer perspectives for the treatment of other AIDS-associated cancers and potentially other virus-related malignancies.

No MeSH data available.


Related in: MedlinePlus

Structure and anti-HIV activity of KPT-185.(A) Structure of KPT-185.(B) Activity of KPT-185 against HIV-1 CXCR4- (IIIB) and CCR5- (BaL) using virus and a multidrug resistant clinical isolate (MDR) in primary peripheral blood lymphocytes. Virus-infected PBMCs were washed 4 days after infection to remove virus in the supernatant and were subsequently incubated with different concentrations of KPT-185 for 1 day. Virus production was analyzed by monitoring the virus-associated p24 core protein in the supernatant by ELISA. Cellular toxicity was measured in parallel using calcein AM live staining and AnnexinV-PI flow cytometry. Error bars represent standard deviations, n = 5.
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f0005: Structure and anti-HIV activity of KPT-185.(A) Structure of KPT-185.(B) Activity of KPT-185 against HIV-1 CXCR4- (IIIB) and CCR5- (BaL) using virus and a multidrug resistant clinical isolate (MDR) in primary peripheral blood lymphocytes. Virus-infected PBMCs were washed 4 days after infection to remove virus in the supernatant and were subsequently incubated with different concentrations of KPT-185 for 1 day. Virus production was analyzed by monitoring the virus-associated p24 core protein in the supernatant by ELISA. Cellular toxicity was measured in parallel using calcein AM live staining and AnnexinV-PI flow cytometry. Error bars represent standard deviations, n = 5.

Mentions: KPT-185 (Fig. 1A) is a SINE compound that effectively and selectively inhibits the XPO1-mediated nuclear export (Neggers et al., 2015). To evaluate the effect of inhibition on HIV replication, we determined the anti-HIV activity of KPT-185 in primary human peripheral blood mononuclear cells (PBMCs). Upon treatment of HIV-infected PBMCs for 24 h, KPT-185 displayed potent anti-HIV activity in these primary cells (IC50: 40 ± 14 nM) (Fig. 1B). The compound proved active against viral strains using the CXCR4 or CCR5 chemokine co-receptor while it caused cytotoxic effects only at concentrations that were 850-fold higher than the active concentration (CC50: 34 ± 13 μM) as measured by calcein AM staining and confirmed by annexin-PI flow cytometry. Furthermore, KPT-185 also suppressed the replication of a clinical isolate (MDR) that was resistant to nucleoside reverse transcriptase inhibitors (NRTI) and protease inhibitors (PI) as well as clinical virus isolates from different subtypes of group M (Table 1). These results illustrate the broad-spectrum anti-HIV activity of XPO1 inhibition.


Human Exportin-1 is a Target for Combined Therapy of HIV and AIDS Related Lymphoma.

Boons E, Vanstreels E, Jacquemyn M, Nogueira TC, Neggers JE, Vercruysse T, van den Oord J, Tamir S, Shacham S, Landesman Y, Snoeck R, Pannecouque C, Andrei G, Daelemans D - EBioMedicine (2015)

Structure and anti-HIV activity of KPT-185.(A) Structure of KPT-185.(B) Activity of KPT-185 against HIV-1 CXCR4- (IIIB) and CCR5- (BaL) using virus and a multidrug resistant clinical isolate (MDR) in primary peripheral blood lymphocytes. Virus-infected PBMCs were washed 4 days after infection to remove virus in the supernatant and were subsequently incubated with different concentrations of KPT-185 for 1 day. Virus production was analyzed by monitoring the virus-associated p24 core protein in the supernatant by ELISA. Cellular toxicity was measured in parallel using calcein AM live staining and AnnexinV-PI flow cytometry. Error bars represent standard deviations, n = 5.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4588406&req=5

f0005: Structure and anti-HIV activity of KPT-185.(A) Structure of KPT-185.(B) Activity of KPT-185 against HIV-1 CXCR4- (IIIB) and CCR5- (BaL) using virus and a multidrug resistant clinical isolate (MDR) in primary peripheral blood lymphocytes. Virus-infected PBMCs were washed 4 days after infection to remove virus in the supernatant and were subsequently incubated with different concentrations of KPT-185 for 1 day. Virus production was analyzed by monitoring the virus-associated p24 core protein in the supernatant by ELISA. Cellular toxicity was measured in parallel using calcein AM live staining and AnnexinV-PI flow cytometry. Error bars represent standard deviations, n = 5.
Mentions: KPT-185 (Fig. 1A) is a SINE compound that effectively and selectively inhibits the XPO1-mediated nuclear export (Neggers et al., 2015). To evaluate the effect of inhibition on HIV replication, we determined the anti-HIV activity of KPT-185 in primary human peripheral blood mononuclear cells (PBMCs). Upon treatment of HIV-infected PBMCs for 24 h, KPT-185 displayed potent anti-HIV activity in these primary cells (IC50: 40 ± 14 nM) (Fig. 1B). The compound proved active against viral strains using the CXCR4 or CCR5 chemokine co-receptor while it caused cytotoxic effects only at concentrations that were 850-fold higher than the active concentration (CC50: 34 ± 13 μM) as measured by calcein AM staining and confirmed by annexin-PI flow cytometry. Furthermore, KPT-185 also suppressed the replication of a clinical isolate (MDR) that was resistant to nucleoside reverse transcriptase inhibitors (NRTI) and protease inhibitors (PI) as well as clinical virus isolates from different subtypes of group M (Table 1). These results illustrate the broad-spectrum anti-HIV activity of XPO1 inhibition.

Bottom Line: Here we report on the dual anti-HIV and anti-PEL effect of targeting a single process common in both diseases.At the same time, SINE caused the nuclear accumulation of p53 tumor suppressor protein as well as inhibition of NF-κB activity in PEL cells resulting in cell cycle arrest and effective apoptosis induction.Our findings provide strong rationale for inhibiting XPO1 as an innovative strategy for the combined anti-retroviral and anti-neoplastic treatment of HIV and PEL and offer perspectives for the treatment of other AIDS-associated cancers and potentially other virus-related malignancies.

View Article: PubMed Central - PubMed

Affiliation: KU Leuven, Department of Microbiology and Immunology, Laboratory of Virology and Chemotherapy, Rega Institute for Medical Research, B-3000 Leuven, Belgium.

ABSTRACT
Infection with HIV ultimately leads to advanced immunodeficiency resulting in an increased incidence of cancer. For example primary effusion lymphoma (PEL) is an aggressive non-Hodgkin lymphoma with very poor prognosis that typically affects HIV infected individuals in advanced stages of immunodeficiency. Here we report on the dual anti-HIV and anti-PEL effect of targeting a single process common in both diseases. Inhibition of the exportin-1 (XPO1) mediated nuclear transport by clinical stage orally bioavailable small molecule inhibitors (SINE) prevented the nuclear export of the late intron-containing HIV RNA species and consequently potently suppressed viral replication. In contrast, in CRISPR-Cas9 genome edited cells expressing mutant C528S XPO1, viral replication was unaffected upon treatment, clearly demonstrating the anti-XPO1 mechanism of action. At the same time, SINE caused the nuclear accumulation of p53 tumor suppressor protein as well as inhibition of NF-κB activity in PEL cells resulting in cell cycle arrest and effective apoptosis induction. In vivo, oral administration arrested PEL tumor growth in engrafted mice. Our findings provide strong rationale for inhibiting XPO1 as an innovative strategy for the combined anti-retroviral and anti-neoplastic treatment of HIV and PEL and offer perspectives for the treatment of other AIDS-associated cancers and potentially other virus-related malignancies.

No MeSH data available.


Related in: MedlinePlus