bioRxiv Science⌕ Search

Biology subjects

Moradpour, S.

Publications and source records attributed to Moradpour, S..

3 recordsLinked to original sources

Non-Canonical Activation of HSF1 Stimulates mTORC1-Mediated Translation in HCMV-Infected Monocytes

Human cytomegalovirus (HCMV) is a major cause of organ disease among immunonaive and immunocompromised individuals. HCMV infection stimulates the survival of normally short-lived circulating monocytes, allowing these blood cells to mediate the dissemination of the virus from the initial point of infection to distant organ sites. We previously showed that HCMV induces a non-canonical phosphorylation of Akt within infected monocytes that activates the stress response transcription factor Heat Shock Factor 1 (HSF1). In this study, we demonstrate that HSF1 is necessary for the survival of HCMV-infected monocytes using both pharmacological and genetic approaches. In contrast, HSF1 inhibition had minimal effect on the viability of uninfected cells, indicating the specific involvement of HSF1 on the survival of infected monocytes. Surprisingly, the aberrant activation of HSF1 by HCMV did not trigger nuclear relocalization, suggesting that HSF1s regulation of monocyte viability occurs within the cytoplasm. Indeed, we found that HCMV-activated, cytoplasmic HSF1 directly binds to mTOR, a critical component of the mTORC1 complex involved in the regulation of mRNA translation. SUnSET (Surface Sensing of Translation) assays revealed HCMV-activated HSF1 increases mRNA translation through mTORC1. Ribosomal profiling identified the increased translation of a selected subset of pro-survival transcripts, including cIAP2, which we validated to selectively stimulate the survival of HCMV-infected monocytes. Taken together, these data demonstrate that the non-canonical activation of HSF1 in infected monocytes drives mTORC1-dependent translation of antiapoptotic transcripts, ensuring the survival and dissemination of infected monocytes. IMPORTANCEHCMV is a primary driver of morbidity and mortality in individuals with compromised or immature immune systems. Spread of HCMV throughout the body relies on the infection of peripheral blood monocytes, which spread the virus to end-organ tissues. However, the naturally short lifespan of monocytes must be overcome to allow for viral spread to occur. Here, we demonstrate that HCMV uniquely regulates the cellular stress response to promote the long-term survival of infected monocytes. Specifically, HCMV activates the cellular stress response transcription factor HSF1 to block the progression of apoptosis. In contrast to traditional heat shock stress where HSF1 translocates into the nucleus to mediate transcription, HCMV infection retains activated HSF1 in the cytoplasm where it binds to mTOR to promote protein synthesis of prosurvival factors necessary for the survival of infected monocytes. Overall, our study provides insight into the complex regulator mechanisms through which HCMV usurps host stress responses to promote viral dissemination.

microbiology↗

Human cytomegalovirus triggered necroptosis is suppressed by sequestration of MLKL in the nucleus of infected monocytes

The systemic spread of human cytomegalovirus (HCMV) is associated with severe morbidity and mortality in immunocompromised and immunonaive patients. Hematogenous dissemination of HCMV to different organ sites is facilitated by peripheral blood monocytes. Circulating monocytes have a short lifespan due, in part, to their intrinsic biological programming to initiate caspase 8-mediated apoptosis upon entry into the circulation from the bone marrow. We previously reported that HCMV extends the lifespan of infected monocytes by blocking procaspase 8 cleavage, yet the precise viral mechanism responsible for suppressing caspase 8 activity remains unknown. Here, we demonstrate that HCMV entry into monocytes rapidly increases the abundance of the antiapoptotic cellular FLICE-like inhibitory protein long (cFLIPL), which prevents procaspase 8 cleavage into active caspase 8. However, others have demonstrated that inhibition of caspase 8 opens a "trapdoor" cell death response termed necroptosis. Accordingly, we found the increased levels of cFLIPL, along with a co-stimulatory signal from toll like receptor 3 (TLR3), activates the receptor-interacting protein kinase 3 (RIPK3) responsible for initiating necroptosis. Despite triggering of the necroptotic cascade within infected monocytes, the final execution of this death pathway is thwarted by nuclear sequestering of mixed lineage kinase domain like pseudokinase (MLKL), the executioner of necroptosis. Together, our data reveal a multitude of countermeasures employed by HCMV to obstruct cellular antiviral death responses within infected monocytes. ImportanceHCMV is highly prevalent in the adult population with a seroprevalence of 50-80% in the United States. Although immunocompetent individuals are generally asymptomatic, HCMV infection can cause multiorgan disease in immunocompromised and immunonaive patients. Peripheral blood monocytes are responsible for the systemic dissemination of HCMV. However, the inherently short lifespan of monocytes combined with the induction of antiviral cellular death responses requires HCMV to circumvents cell death pathways to allow for viral spread. In this work, we show that HCMV induces cFLIPL levels to inhibit caspase 8-mediated apoptosis. However, the inhibition of apoptosis, combined with TLR3 activation, triggers a secondary cell death pathway termed necroptosis. As a countermeasure to block necroptosis, HCMV sequesters MLKL within the nucleus of infected monocytes. Defining the precise mechanisms through which HCMV stimulates survival will provide insight into novel therapeutics able to target infected monocytes.

microbiology↗

B cell receptor dependent enhancement of dengue virus infection

Dengue virus (DENV) is the causative agent of dengue, a mosquito-borne disease that represents a significant and growing public health burden around the world. A unique pathophysiological feature of dengue is immune-mediated enhancement, wherein preexisting immunity elicited by a primary infection can enhance the severity of a subsequent infection by a heterologous DENV serotype. A leading mechanistic explanation for this phenomenon is antibody dependent enhancement (ADE), where sub-neutralizing concentrations of DENV-specific IgG antibodies facilitate entry of DENV into Fc{gamma}R expressing cells such as monocytes, macrophages, and dendritic cells. Accordingly, this model posits that phagocytic mononuclear cells are the primary reservoir of DENV. However, analysis of samples from individuals experiencing acute DENV infection reveals that B cells are the largest reservoir of infected circulating cells, representing a disconnect in our understanding of immune-mediated DENV tropism. In this study, we demonstrate that the expression of a DENV-specific B cell receptor (BCR) renders cells highly susceptible to DENV infection, with the infection-enhancing activity of the membrane-restricted BCR correlating with the ADE potential of the IgG version of the antibody. In addition, we observed that the frequency of DENV-infectable B cells increases in previously flavivirus-naive volunteers after a primary DENV infection. These findings suggest that BCR-dependent infection of B cells is a novel mechanism immune-mediated enhancement of DENV-infection.

immunology↗