IMMU-22. EARLY RELEASE OF TYPE I INTERFERON DRIVES BRAIN CANCER IMMUNOGENICITY AND RESPONSE TO IMMUNOTHERAPY

Abstract BACKGROUND Molecular drivers of cancer immunogenicity in brain tumors are still being unraveled. While BATF3 expression, STING, and interferon response factors (IRFs) are necessary for cancer immunogenicity, the presence of type I interferon (IFN-I) is contextual having been reported to elicit both anti-tumoral and pro-tumoral effects. A better understanding of IFN-I signaling mechanisms is necessary to elucidate drivers of brain cancer immunogenicity and resistance. OBJECTIVE We sought to assess the role of IFN-I signaling in brain tumor immunogenicity and response to immune checkpoint inhibitors (ICIs) in ICI sensitive brain tumor models (i.e. GL261). We then sought to develop strategies to reset IFN-I signaling in ICI resistant brain tumor models (i.e. KR158b). METHODS To reset IFN-I signaling in immunologically ‘cold’ tumors unresponsive to ICIs, we developed lipid-nanoparticles (NPs) to deliver mRNA payloads to the brain tumor microenvironment (TME). RESULTS In immune-sensitive GL261 tumors, we showed that early release of IFN-I unlocks cancer immunogenicity and ICI response. Blockade of IFN-I during tumorigenesis (within 24h, but not days later) increases tumorigenicity and abrogates ICI activity in sensitive tumors. In ICI resistant KR158b tumors, we show that systemic administration of tumor-derived RNA-NPs localize to myeloid cells within the TME for simultaneous activation of multiple innate pathways including BATF3 (necessary for effector DCs), IRF5 (necessary for M2 to M1 macrophage reprograming), and IRF7 (necessary for IFN-I production). These RNA-NPs induce near-immediate release of IFN-I (within hours), reprogram the brain TME in an IFNAR1 (IFN-I receptor) dependent manner, and elicit significant anti-KR158b efficacy as a monotherapy. Following IFNAR1 blockade, RNA-NP mediated anti-tumor efficacy was abrogated. We demonstrated safety of tumor-specific RNA-NPs (derived from KR158b) in acute/chronic GLP toxicity studies without normal-brain cross-reactivity, and confirmed feasibility/safety and immunologic activity in large-animal studies. FUTURE DIRECTIONS We have since received FDA-IND approval for first-in-human trials (IND#BB-19304) in glioblastoma patients.

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The Type I Interferon Response Determines Differences in Choroid Plexus Susceptibility between Newborns and Adults in Herpes Simplex Virus Encephalitis

mBio ◽

10.1128/mbio.00437-16 ◽

2016 ◽

Vol 7 (2) ◽

Cited By ~ 19

Author(s):

Douglas R. Wilcox ◽

Stephen S. Folmsbee ◽

William J. Muller ◽

Richard Longnecker

Keyword(s):

Choroid Plexus ◽

Type I Interferon ◽

Adult Brain ◽

Interferon Response ◽

Type I ◽

Β Receptor ◽

Simplex Virus ◽

The Brain ◽

Hsv 1

ABSTRACTNewborns are significantly more susceptible to severe viral encephalitis than adults, with differences in the host response to infection implicated as a major factor. However, the specific host signaling pathways responsible for differences in susceptibility and neurologic morbidity have remained unknown. In a murine model of HSV encephalitis, we demonstrated that the choroid plexus (CP) is susceptible to herpes simplex virus 1 (HSV-1) early in infection of the newborn but not the adult brain. We confirmed susceptibility of the CP to HSV infection in a human case of newborn HSV encephalitis. We investigated components of the type I interferon (IFN) response in the murine brain that might account for differences in cell susceptibility and found that newborns have a dampened interferon response and significantly lower basal levels of the alpha/beta interferon (IFN-α/β) receptor (IFNAR) than do adults. To test the contribution of IFNAR to restricting infection from the CP, we infected IFNAR knockout (KO) adult mice, which showed restored CP susceptibility to HSV-1 infection in the adult. Furthermore, reduced IFNAR levels did not account for differences we found in the basal levels of several other innate signaling proteins in the wild-type newborn and the adult, including protein kinase R (PKR), that suggested specific regulation of innate immunity in the developing brain. Viral targeting of the CP, a region of the brain that plays a critical role in neurodevelopment, provides a link between newborn susceptibility to HSV and long-term neurologic morbidity among survivors of newborn HSV encephalitis.IMPORTANCECompared to adults, newborns are significantly more susceptible to severe disease following HSV infection. Over half of newborn HSV infections result in disseminated disease or encephalitis, with long-term neurologic morbidity in 2/3 of encephalitis survivors. We investigated differences in host cell susceptibility between newborns and adults that contribute to severe central nervous system disease in the newborn. We found that, unlike the adult brain, the newborn choroid plexus (CP) was susceptible early in HSV-1 infection. We demonstrated that IFN-α/β receptor levels are lower in the newborn brain than in the adult brain and that deletion of this receptor restores susceptibility of the CP in the adult brain. The CP serves as a barrier between the blood and the cerebrospinal fluid and plays a role in proper neurodevelopment. Susceptibility of the newborn choroid plexus to HSV-1 has important implications in viral spread to the brain and, also, in the neurologic morbidity following HSV encephalitis.

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Molecular Insight Into the IRE1α-Mediated Type I Interferon Response Induced by Proteasome Impairment in Myeloid Cells of the Brain

Frontiers in Immunology ◽

10.3389/fimmu.2019.02900 ◽

2019 ◽

Vol 10 ◽

Cited By ~ 3

Author(s):

Maja Studencka-Turski ◽

Gonca Çetin ◽

Heike Junker ◽

Frédéric Ebstein ◽

Elke Krüger

Keyword(s):

Type I Interferon ◽

Myeloid Cells ◽

Interferon Response ◽

Type I ◽

The Brain ◽

Insight Into

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The brain parenchyma has a type I interferon response that can limit virus spread

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1618157114 ◽

2016 ◽

Vol 114 (1) ◽

pp. E95-E104 ◽

Cited By ~ 25

Author(s):

Eugene Drokhlyansky ◽

Didem Göz Aytürk ◽

Timothy K. Soh ◽

Ryan Chrenek ◽

Elaine O’Loughlin ◽

...

Keyword(s):

Immune Response ◽

Innate Immune Response ◽

Type I Interferon ◽

Brain Parenchyma ◽

Innate Immune ◽

Interferon Response ◽

Type I ◽

Virus Spread ◽

Viral Spread ◽

The Brain

The brain has a tightly regulated environment that protects neurons and limits inflammation, designated “immune privilege.” However, there is not an absolute lack of an immune response. We tested the ability of the brain to initiate an innate immune response to a virus, which was directly injected into the brain parenchyma, and to determine whether this response could limit viral spread. We injected vesicular stomatitis virus (VSV), a transsynaptic tracer, or naturally occurring VSV-derived defective interfering particles (DIPs), into the caudate–putamen (CP) and scored for an innate immune response and inhibition of virus spread. We found that the brain parenchyma has a functional type I interferon (IFN) response that can limit VSV spread at both the inoculation site and among synaptically connected neurons. Furthermore, we characterized the response of microglia to VSV infection and found that infected microglia produced type I IFN and uninfected microglia induced an innate immune response following virus injection.

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Bud13 Promotes a Type I Interferon Response by Countering Intron Retention in Irf7

10.1101/443820 ◽

2018 ◽

Author(s):

Luke S. Frankiw ◽

Devdoot Majumdar ◽

Christian Burns ◽

Annie Moradian ◽

Michael J. Sweredoski ◽

...

Keyword(s):

Rna Binding ◽

Type I Interferon ◽

Intron Retention ◽

Global Analysis ◽

Interferon Response ◽

Type I ◽

Dependent Manner ◽

Protein Levels ◽

Expression Of Genes ◽

Gene Expression Control

SUMMARYIntron retention (IR) has emerged as an important mechanism of gene expression control. Despite this, the factors that control IR events remain poorly understood. We observed consistent IR in one intron of the Irf7 gene and identified Bud13 as an RNA-binding protein that acts at this intron to increase the amount of successful splicing. Deficiency in Bud13 led to increased IR, decreased mature Irf7 transcript and protein levels, and consequently to a dampened type I interferon response. This impairment of Irf7 production in Bud13-deficient cells compromised their ability to withstand VSV infection. Global analysis of Bud13 knockdown and BUD13 cross-linking to RNA revealed a subset of introns that share many characteristics with the one found in Irf7 and are spliced in a Bud13-dependent manner. Deficiency of Bud13 led to decreased mature transcript from genes containing such introns. Thus, by acting as an antagonist to IR, Bud13 facilitates the expression of genes at which IR occurs.

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A type I interferon response defines a conserved microglial state required for effective phagocytosis

10.1101/2021.04.29.441889 ◽

2021 ◽

Author(s):

Leah C Dorman ◽

Phi T Nguyen ◽

Caroline C Escoubas ◽

Ilia D Vainchtein ◽

Yinghong Xiao ◽

...

Keyword(s):

Type I Interferon ◽

Meta Analysis ◽

Transcriptional Profiling ◽

Innate Immune ◽

Interferon Response ◽

Type I ◽

Whole Cells ◽

Microglial Phagocytosis ◽

The Brain

Microglia, the innate immune cells of the brain, are exquisitely sensitive to dynamic changes in the brain environment. We used single cell RNA sequencing to define glial responses in the early postnatal somatosensory cortex after partial whisker lesion, revealing transcriptomic shifts in both astrocytes and microglia during the resulting topographic remapping. The most distinct change was the emergence of a type I interferon (IFN-I) responsive microglia population that was rare in the resting cortex but expanded 20-fold after whisker deprivation. The top gene candidate in this cluster, Ifitm3, marked a conserved but transient subset of microglia that were in the process of phagocytosing whole cells. IFITM3 protein identified this subset in vivo, where it was enriched in early microglial phagosomes. Loss of canonical IFN-I signaling in Ifnar1-/- animals resulted in abnormal 'bubble' microglia with deficient phagolysosomal processing. In a meta-analysis of transcriptomes, we identified the IFN-I signature in microglia across a range of pathologies. We identified phagocytic IFITM3+ microglia in two murine disease models: SARS-CoV-2 infection and Alzheimer's Disease. These data reveal the potential of transcriptional profiling after defined perturbation to elicit transient microglial states, and identify a novel role for IFN-I signaling in regulating microglial phagocytosis.

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Suppression of Type I Interferon Signaling by E1A via RuvBL1/Pontin

Journal of Virology ◽

10.1128/jvi.02484-16 ◽

2017 ◽

Vol 91 (8) ◽

Cited By ~ 8

Author(s):

Oladunni Olanubi ◽

Jasmine Rae Frost ◽

Sandi Radko ◽

Peter Pelka

Keyword(s):

Gene Expression ◽

Cellular Response ◽

Type I Interferon ◽

Early Gene ◽

Interferon Response ◽

Type I ◽

Dependent Manner ◽

Interferon Signaling ◽

Interferon Pathway ◽

Regulated Promoters

ABSTRACT Suppression of interferon signaling is of paramount importance to a virus. Interferon signaling significantly reduces or halts the ability of a virus to replicate; therefore, viruses have evolved sophisticated mechanisms that suppress activation of the interferon pathway or responsiveness of the infected cell to interferon. Adenovirus has multiple modes of inhibiting the cellular response to interferon. Here, we report that E1A, previously shown to regulate interferon signaling in multiple ways, inhibits interferon-stimulated gene expression by modulating RuvBL1 function. RuvBL1 was previously shown to affect type I interferon signaling. E1A binds to RuvBL1 and is recruited to RuvBL1-regulated promoters in an interferon-dependent manner, preventing their activation. Depletion of RuvBL1 impairs adenovirus growth but does not appear to significantly affect viral protein expression. Although RuvBL1 has been shown to play a role in cell growth, its depletion had no effect on the ability of the virus to replicate its genome or to drive cells into S phase. E1A was found to bind to RuvBL1 via the C terminus of E1A, and this interaction was important for suppression of interferon-stimulated gene transcriptional activation and recruitment of E1A to interferon-regulated promoters. Here, we report the identification of RuvBL1 as a new target for adenovirus in its quest to suppress the interferon response. IMPORTANCE For most viruses, suppression of the interferon signaling pathway is crucial to ensure a successful replicative cycle. Human adenovirus has evolved several different mechanisms that prevent activation of interferon or the ability of the cell to respond to interferon. The viral immediate-early gene E1A was previously shown to affect interferon signaling in several different ways. Here, we report a novel mechanism reliant on RuvBL1 that E1A uses to prevent activation of interferon-stimulated gene expression following infection or interferon treatment. This adds to the growing knowledge of how viruses are able to inhibit interferon and identifies a novel target used by adenovirus for modulation of the cellular interferon pathway.

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Aging-induced type I interferon response at the choroid plexus negatively affects brain function

Science ◽

10.1126/science.1252945 ◽

2014 ◽

Vol 346 (6205) ◽

pp. 89-93 ◽

Cited By ~ 239

Author(s):

Kuti Baruch ◽

Aleksandra Deczkowska ◽

Eyal David ◽

Joseph M. Castellano ◽

Omer Miller ◽

...

Keyword(s):

Cognitive Decline ◽

Choroid Plexus ◽

Brain Function ◽

Type I Interferon ◽

Negative Influence ◽

Hippocampal Neurogenesis ◽

Interferon Response ◽

Type I ◽

Aged Mice ◽

The Brain

Aging-associated cognitive decline is affected by factors produced inside and outside the brain. By using multiorgan genome-wide analysis of aged mice, we found that the choroid plexus, an interface between the brain and the circulation, shows a type I interferon (IFN-I)–dependent gene expression profile that was also found in aged human brains. In aged mice, this response was induced by brain-derived signals, present in the cerebrospinal fluid. Blocking IFN-I signaling within the aged brain partially restored cognitive function and hippocampal neurogenesis and reestablished IFN-II–dependent choroid plexus activity, which is lost in aging. Our data identify a chronic aging-induced IFN-I signature, often associated with antiviral response, at the brain’s choroid plexus and demonstrate its negative influence on brain function, thereby suggesting a target for ameliorating cognitive decline in aging.

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Whole-brain fluorescence-MRI coregistration for precise anatomical mapping of virus infection

10.1101/2021.11.16.468804 ◽

2021 ◽

Author(s):

Nunya Chotiwan ◽

Stefanie M.A. Willekens ◽

Erin Schexnaydre ◽

Max Hahn ◽

Federico Morini ◽

...

Keyword(s):

Virus Infection ◽

Type I Interferon ◽

Ex Vivo ◽

Interferon Response ◽

Type I ◽

Virus Infections ◽

Whole Brain ◽

Neurotropic Virus ◽

Langat Virus ◽

The Brain

Neurotropic virus infections cause tremendous disease burden. Methods visualizing infection in the whole brain remain unavailable which greatly impedes understanding of viral neurotropism and pathogenesis. We devised an approach to visualize the distribution of neurotropic virus infection in whole mouse brain ex vivo. Optical projection tomography (OPT) signal was coregistered with a unique magnetic resonance imaging (MRI) brain template, enabling precise anatomical mapping of viral distribution, and the effect of type I interferon on distribution of infection was analyzed. Guided by OPT-MR, we show that Langat virus specifically targets sensory brain systems and the lack of type I interferon response results in an anatomical shift in infection patterns in the brain. We confirm this regional tropism, observed with whole brain OPT-MRI, by confocal and electron microscopy to provide unprecedented insight into viral neurotropism. This approach can be applied to any fluorescently labeled target in the brain.

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The Second-Generation XPO1 Inhibitor Eltanexor Inhibits Human Cytomegalovirus (HCMV) Replication and Promotes Type I Interferon Response

Frontiers in Microbiology ◽

10.3389/fmicb.2021.675112 ◽

2021 ◽

Vol 12 ◽

Author(s):

Yueyan Liao ◽

Xiangyu Ke ◽

Tianyi Deng ◽

Qingsong Qin

Keyword(s):

Human Cytomegalovirus ◽

Viral Entry ◽

Nuclear Export ◽

Second Generation ◽

Type I Interferon ◽

Opportunistic Pathogen ◽

Interferon Response ◽

Type I ◽

Dependent Manner ◽

Hcmv Replication

Human cytomegalovirus (HCMV) is a ubiquitous opportunistic pathogen and can be life-threatening for immunocompromised individuals. There is currently no available vaccine for the prevention of HCMV- associated diseases and most of the available antiviral drugs that target viral DNA synthesis become ineffective in treating HCMV mutants that arise after long-term use in immunocompromised patients. Here, we examined the effects of Eltanexor, a second-generation selective inhibitor of nuclear export (SINE), on HCMV replication. Eltanexor effectively inhibits HCMV replication in human foreskin fibroblasts in a dose-dependent manner. Eltanexor does not significantly inhibit viral entry and nuclear import of viral genomic DNA, but rather suppress the transcript and protein levels of viral immediate-early (IE), early (E) and late (L) genes, and abolishes the production of infectious virions. We further found Eltanexor treatment promotes proteasome-mediated degradation of XPO1, which contributes to the nuclear retention of interferon regulatory factor 3 (IRF-3), resulting in increased expression of type I interferon as well as interferon stimulating genes ISG15 and ISG54. This study reveals a novel antiviral mechanism of Eltanexor which suggests it has potential to inhibit a broad spectrum of viral pathogens.

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Porcine Epidemic Diarrhea Virus Infection Inhibits Interferon Signaling by Targeted Degradation of STAT1

Journal of Virology ◽

10.1128/jvi.01091-16 ◽

2016 ◽

Vol 90 (18) ◽

pp. 8281-8292 ◽

Cited By ~ 34

Author(s):

Longjun Guo ◽

Xiaolei Luo ◽

Ren Li ◽

Yunfei Xu ◽

Jian Zhang ◽

...

Keyword(s):

Virus Infection ◽

Porcine Epidemic Diarrhea Virus ◽

Molecular Mechanisms ◽

Type I Interferon ◽

Infection Type ◽

Interferon Response ◽

Type I ◽

Dependent Manner ◽

Diarrhea Virus ◽

Porcine Epidemic Diarrhea

ABSTRACTPorcine epidemic diarrhea virus (PEDV) is a worldwide-distributed alphacoronavirus, but the pathogenesis of PEDV infection is not fully characterized. During virus infection, type I interferon (IFN) is a key mediator of innate antiviral responses. Most coronaviruses develop some strategy for at least partially circumventing the IFN response by limiting the production of IFN and by delaying the activation of the IFN response. However, the molecular mechanisms by which PEDV antagonizes the antiviral effects of interferon have not been fully characterized. Especially, how PEDV impacts IFN signaling components has yet to be elucidated. In this study, we observed that PEDV was relatively resistant to treatment with type I IFN. Western blot analysis showed that STAT1 expression was markedly reduced in PEDV-infected cells and that this reduction was not due to inhibition of STAT1 transcription. STAT1 downregulation was blocked by a proteasome inhibitor but not by an autophagy inhibitor, strongly implicating the ubiquitin-proteasome targeting degradation system. Since PEDV infection-induced STAT1 degradation was evident in cells pretreated with the general tyrosine kinase inhibitor, we conclude that STAT1 degradation is independent of the IFN signaling pathway. Furthermore, we report that PEDV-induced STAT1 degradation inhibits IFN-α signal transduction pathways. Pharmacological inhibition of STAT1 degradation rescued the ability of the host to suppress virus replication. Collectively, these data show that PEDV is capable of subverting the type I interferon response by inducing STAT1 degradation.IMPORTANCEIn this study, we show that PEDV is resistant to the antiviral effect of IFN. The molecular mechanism is the degradation of STAT1 by PEDV infection in a proteasome-dependent manner. This PEDV infection-induced STAT1 degradation contributes to PEDV replication. Our findings reveal a new mechanism evolved by PEDV to circumvent the host antiviral response.

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