An iPSC-Derived Myeloid Lineage Model of Herpes Virus Latency and Reactivation

Poole, Emma; Huang, Ching-Kuei; Forbester, Jessica L.; Shnayder, Miri; Nachshon, Aharon; Kweider, Baraa; Basaj, Anna; Smith, Daniel P.; Jackson, Sarah E.; Liu, Bin; Shih, Joy; Kiskin, Fedir; Roche, Kathryn L.; Murphy, Eain A.; Wills, Mark R.; Morrell, Nicholas W.; Dougan, Gordon; Stern-Ginossar, Noam; Amer, Radgonde

doi:10.3389/fmicb.2019.02233

Cited by 18 publications

(19 citation statements)

References 49 publications

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“…Previous studies have shown efficacy of I-BETs in various models of human myeloid leukemia, and many inhibitors are now in clinical development (phases I, II, and III) as single treatments of both oncology and nononcology diseases ( 66 ). Our work also shows that I-BETs can initiate IE gene expression from HCMV latently infected CD34+ cells, which are good models for HCMV latent infection ( 69 ), whereas HDACis such as CHR-4487 (a myeloid-selective HDACi) can have differential effects on CD34+ models with little response in Kasumi-3 cells, consistent with others’ previous work ( 70 ). Hence, with good compound penetrance and known immune surveillance in bone marrow, treatment with I-BETs like GSK726 may also allow targeting of latently infected bone-marrow resident pluripotent CD34+ progenitor cells.…”

Section: Discussionsupporting

confidence: 91%

Bromodomain proteins regulate human cytomegalovirus latency and reactivation allowing epigenetic therapeutic intervention

Groves

Jackson

Poole

et al. 2021

Proc. Natl. Acad. Sci. U.S.A.

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Reactivation of human cytomegalovirus (HCMV) from latency is a major health consideration for recipients of stem-cell and solid organ transplantations. With over 200,000 transplants taking place globally per annum, virus reactivation can occur in more than 50% of cases leading to loss of grafts as well as serious morbidity and even mortality. Here, we present the most extensive screening to date of epigenetic inhibitors on HCMV latently infected cells and find that histone deacetylase inhibitors (HDACis) and bromodomain inhibitors are broadly effective at inducing virus immediate early gene expression. However, while HDACis, such as myeloid-selective CHR-4487, lead to production of infectious virions, inhibitors of bromodomain (BRD) and extraterminal proteins (I-BETs), including GSK726, restrict full reactivation. Mechanistically, we show that BET proteins (BRDs) are pivotally connected to regulation of HCMV latency and reactivation. Through BRD4 interaction, the transcriptional activator complex P-TEFb (CDK9/CycT1) is sequestered by repressive complexes during HCMV latency. Consequently, I-BETs allow release of P-TEFb and subsequent recruitment to promoters via the superelongation complex (SEC), inducing transcription of HCMV lytic genes encoding immunogenic antigens from otherwise latently infected cells. Surprisingly, this occurs without inducing many viral immunoevasins and, importantly, while also restricting viral DNA replication and full HCMV reactivation. Therefore, this pattern of HCMV transcriptional dysregulation allows effective cytotoxic immune targeting and killing of latently infected cells, thus reducing the latent virus genome load. This approach could be safely used to pre-emptively purge the virus latent reservoir prior to transplantation, thereby reducing HCMV reactivation-related morbidity and mortality.

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Section: Discussionsupporting

confidence: 91%

Bromodomain proteins regulate human cytomegalovirus latency and reactivation allowing epigenetic therapeutic intervention

Groves

Jackson

Poole

et al. 2021

Proc. Natl. Acad. Sci. U.S.A.

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“…TB40/E mCherry -US28-3XFLAG and TB40/E mCherry -US28Δ have been described previously (19). TB40/E gfp (82) and TB40/E BAC4 SV40 mCherry IE2-2A-GFP (83) were kind gifts from E. A. Murphy, SUNY Upstate Medical University. TB40/E BAC4 IE2-eYFP has been described previously (60, 84).…”

Section: Methodsmentioning

confidence: 99%

Interferon-Responsive Genes Are Targeted during the Establishment of Human Cytomegalovirus Latency

Elder

Krishna

Williamson

et al. 2019

mBio

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Human cytomegalovirus (HCMV) is a ubiquitous herpesvirus which infects 50 to 100% of humans worldwide. HCMV causes a lifelong subclinical infection in immunocompetent individuals but is a serious cause of mortality and morbidity in the immunocompromised and neonates. In particular, reactivation of HCMV in the transplant setting is a major cause of transplant failure and related disease. Therefore, a molecular understanding of HCMV latency and reactivation could provide insights into potential ways to target the latent viral reservoir in at-risk patient populations.

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“…TB40/E mCherry -US28-3XFLAG and TB40/E mCherry -US28Δ have been described previously 19 . TB40/E gfp 47 and TB40/E BAC4 SV40 mCherry IE2-2A-GFP 48 were kind gifts from E.A. Murphy, SUNY Upstate Medical University.…”

Section: Methodsmentioning

confidence: 99%

Interferon-responsive genes are targeted during the establishment of human cytomegalovirus latency

Krishna

Williamson

Lim

et al. 2019

Preprint

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12Human cytomegalovirus (HCMV) latency is an active process which remodels the latently infected 13 cell to optimise latent carriage and reactivation. This is achieved, in part, through the expression of 14 viral genes, including the G-protein coupled receptor US28. Here, we use an unbiased proteomic 15 screen to assess changes in host proteins induced by US28, revealing that interferon-inducible genes 16 are downregulated by US28. We validate that MHC Class II and two PYHIN proteins, MNDA and 17 IFI16, are downregulated during experimental latency in primary human CD14 + monocytes. We find 18 that IFI16 is targeted rapidly during the establishment of latency in a US28-dependent manner, but 19 only in undifferentiated myeloid cells, a natural site of latent carriage. Finally, by overexpressing 20 IFI16, we show that IFI16 can activate the viral major immediate early promoter and immediate early 21 2 gene expression during latency via NF-κB, a function which explains why downregulation of IFI16 22 during latency is advantageous for the virus. 23 Importance 24 Human cytomegalovirus (HCMV) is a ubiquitous herpesvirus which infects 50-100% of humans 25 worldwide. HCMV causes a lifelong subclinical infection in immunocompetent individuals, but is a 26 serious cause of mortality and morbidity in the immunocompromised and in neonates. In particular, 27 reactivation of HCMV in the transplant setting is a major cause of transplant failure and related 28 disease. Therefore, a molecular understanding of HCMV latency and reactivation could provide 29 insights into potential ways to target the latent viral reservoir in at-risk patient populations. 30 31 both host and viral factors 18 . One viral factor that suppresses MIEP activity is the G-protein coupled 46 receptor (GPCR) US28, a virally encoded chemokine receptor homologue, which is expressed de 47 novo during latency as well as being delivered to cells with the incoming virion 19-24 and this 48 incoming viral US28 is functional 25 . US28 modulates the signalling pathways of early myeloid cells; it 49 attenuates MAP kinases, NF-κB, and c-fos, whilst activating STAT3 and iNOS 21,23,25 . All of these 50 contribute to the repression of MIEP activity. This US28-mediated signalling is so critical to latency 51 that US28-deleted viruses, or the loss of G-protein coupling by the US28 mutant R129A, result in lytic 52 infection of undifferentiated myeloid cells 19,21,23,25 . Furthermore, when examined, these US28-53 mediated effects on cell signalling did not occur during lytic infection or in permissive cells 21 , 54 implying that US28 represses the MIEP during latency but does not impair reactivation following 55 cellular differentiation. This is reflective of the cell type-specific nature of US28-mediated signalling 56 21,26 . 57Since US28 can modulate all these pathways and control the MIEP, we hypothesised that US28 58 would also cause changes in host protein expression. Here, we perform a proteomic screen 59 comparing host cell protein abundance in myelomonocytic TH...

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An iPSC-Derived Myeloid Lineage Model of Herpes Virus Latency and Reactivation

Cited by 18 publications

References 49 publications

Bromodomain proteins regulate human cytomegalovirus latency and reactivation allowing epigenetic therapeutic intervention

Bromodomain proteins regulate human cytomegalovirus latency and reactivation allowing epigenetic therapeutic intervention

Interferon-Responsive Genes Are Targeted during the Establishment of Human Cytomegalovirus Latency

Interferon-responsive genes are targeted during the establishment of human cytomegalovirus latency

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