Manipulation of inflammasomes and NF-kB signaling in human myeloid cells by Myxom

Myxom 操纵人骨髓细胞中的炎症小体和 NF-kB 信号传导

• 批准号: 9382931
• 负责人: Grant McFadden
• 金额: $ 38.63万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9382931
• 关键词: Adaptor Signaling Protein; Address; Allografting; Apoptosis; Area; Binding; Binding Proteins; CASP1 gene; Carcinoma in Situ; Cell Line; Cells; Clinical; Complex; Cytokine Signaling; Dendritic Cells; Disease; Disease model; Engineering; European; Exhibits; Family; Family member; Human; Immune; Immune Evasion; Immune signaling; Immunocompetent; Immunocompromised Host; In Situ; Individual; Infection; Inflammasome; Inflammatory; Innate Immune Response; Interferons; Interleukin-1 beta; Interleukin-18; Investigation; Knockout Mice; Knowledge; Laboratories; Lagomorpha; MAP Kinase Gene; MAP2K1 gene; MAPK3 gene; Malignant Neoplasms; Mediating; Modeling; Molecular; Multiple Myeloma; Mus; Myelogenous; Myeloid Cells; Myeloid Leukemia; Myxoma virus; NF-kappa B; Nature; Oncolytic; Oncolytic viruses; Oryctolagus cuniculus; Pathway interactions; Penetration; Permeability; Pharmaceutical Preparations; Phosphotransferases; Play; Poxviridae; Process; Proteins; Rattus; Recombinants; Regulation; Reporting; Role; Sentinel; Signal Pathway; Signal Transduction; TNF gene; Testing; Therapeutic; Transducers; Transplantation; Transplanted tissue; Tropism; Variant; Vascular Diseases; Viral; Viral Proteins; Virotherapy; Virus; Virus Diseases; ataxia telangiectasia mutated protein; base; cancer cell; cell type; cytokine; immunoregulation; in vivo; macrophage; marenostrin; monocyte; mouse model; mutant; novel therapeutic intervention; oncolysis; oncolytic virotherapy; permissiveness; public health relevance; receptor; response; sensor; tool; tumor; virus tropism

DESCRIPTION (provided by applicant): In this proposal, we will determine how key innate immune response pathways to viral infection are altered in human cancer cells, particularly by comparing primary innate immune sentinel myeloid cells to their transformed counterparts. We will investigate the mechanistic roles of virus sensing pathways and their linkage to downstream innate immune responses via inflammasomes and NF-κB signaling in determining the tropism and sensing of myxoma virus (MYXV), a rabbit specific poxvirus that also exhibits the capacity to infect a wide spectrum of human cancer cells. We will investigate MYXV sensing and infection of human myeloid cells, especially monocyte-derived macrophages and dendritic cells. We will examine the molecular mechanisms used by MYXV-encoded pyrin domain (PYD)-containing host range protein M013 to co-regulate two distinct innate immune pathways of human myeloid cells, particularly as mediated by inflammasomes and by the NF-κB-mediated inflammatory signaling responses. We have reported that M013 binds the cellular adaptor protein ASC-1 (apoptosis-associated speck-like protein containing a CARD) of various inflammasome complexes and NF-κB1/p105 of NF-κB signaling pathway. Investigation of this unique co-regulation will have potential clinical utility for developing drugs having the capacity o target multiple inflammatory signaling pathways. Finally, we will take advantage of our knowledge of regulation of the inflammasome and NF-κB pathways to study the mechanism(s) of oncolysis by MYXV using myeloid leukemia models. Using knock-out mice for key innate immune pathways and MYXV constructed mutated at the M013 locus, we will study the role of these pathways in MYXV virotherapy for myeloid cancer. The three overlapping areas to be investigated in this proposal are: Aim 1. Study the alteration of sensing pathways due to transformation of human myeloid cells and its impact on MYXV tropism: We will determine the role of RLR and TLR signaling in inducing type I IFN and pro-inflammatory cytokines in human myeloid cells that are permissive for MYXV (eg transformed myeloid cell lines like THP-1) or nonpermissive (eg primary human monocytes/macrophages); determine the role of NLRP3 inflammasome components and NF-κB1 in the synthesis of precursor IL-1ß and IL-18 and release of these mature cytokines in response to MYXV infection; study the co-ordinating upstream role of virus-induced MAPK, in particular the role of MEK1/2-ERK1/2 kinases, in co-inducing both the type I IFN and pro-inflammatory cytokine signaling pathways in response to MYXV infection; and identify specifically which cellular sensor/transducer components become dysfunctional in transformed human myeloid cells. Aim 2. Create engineered variants of the PYD-containing protein M013 that differentially regulate the inflammasome and NF-κB signaling pathways and test these M013 constructs in an in vivo caspase 1- dependent rat transplant vascular disease model: We will generate M013 mutants that selectively inhibit either the inflammasome or NF-κB pathways by interaction with only ASC-1 or NF-κB1 and exploit these mutants to construct recombinant MYXVs to test their effects in the myeloid cells described above; we will also test M013 variants engineered to allow intracellular penetration in a rat allograft transplant vascular disease (TVD) model that has been shown to be regulated by inflammasome-mediated activation of caspase 1. Aim 3. Elucidate the roles of host inflammasomes and NF-κB signaling in MYXV-induced oncolytic clearance of myeloid cancer in situ: We will compare the roles of key inflammasome and NF-κB components (ie ASC-1, caspase-1, NLRP3 and NF-κB1) in MYXV-induced anti-tumor responses and clearance of human (eg THP-1) or murine (eg RAW426.7) myeloid leukemias in NSG immunocompromised mice and in C57BL/6-based immunocompetent mouse models, respectively; study the role of M013 variants in selective knockout mice using the M013KO vs wild-type MYXV, and the M013 variant-expressing viruses that selectively inhibit only the inflammasome or NF-κB pathways.

描述（由适用提供）：在此提案中，我们将确定人类癌细胞中关键的先天免疫响应途径如何改变了病毒感染的途径，尤其是通过比较原发性先天性免疫素髓样细胞与其转化的对应物。我们将研究病毒敏感性途径的机理作用及其通过炎症和NF-κB信号与下游先天免疫响应的联系在确定粘液瘤病毒（Myxv）的性质和敏感性（一种兔子特异性蛇毒）的敏感性方面，这也表现出具有人类癌症癌细胞的能力。我们将研究人髓样细胞的MyXV敏感性和感染，尤其是单核细胞衍生的巨噬细胞和树突状细胞。我们将研究由MyXV编码的吡啶结构域（PYD）含有宿主范围蛋白M013使用的分子机制，以共同调节人髓样细胞的两个不同的先天免疫途径，特别是由炎症和NF-κB介导的炎症信号传导反应。我们报道了M013结合了各种炎性体复合物的细胞适配器蛋白ASC-1（含有凋亡相关的含有卡片样蛋白）和NF-κB信号通路的NF-κB1/p105。对这种独特的共同调节的研究将具有潜在的临床实用性，以开发具有o目标多重炎症信号通路的能力的药物。最后，我们将利用对炎性体和NF-κB途径的调节知识来研究使用髓样白血病模型来研究MyxV癌变的机制。使用在M013基因座进行突变的关键先天免疫途径的敲除小鼠，我们将研究这些途径在MyXV病毒疗法中的髓样癌中的作用。 The three overlapping areas to be investigated in This proposal are: Aim 1. Study the alteration of sensing pathways due to transformation of human myeloid cells and its impact on MYXV tropism: We will determine the role of RLR and TLR signaling in induced type I IFN and pro-inflammatory cytokines in human myeloid cells that are permissive for MYXV (eg transformed myeloid cell lines like THP-1) or nonpermissive （例如原代人单核细胞/巨噬细胞）；确定NLRP3炎性组成分和NF-κB1在响应MYXV感染的响应的前体IL-1ß和IL-18合成中的作用以及这些成熟细胞因子的释放；研究病毒诱导的MAPK的协调上游作用，尤其是MEK1/2-ERK1/2激酶的作用，在共同引起的I型IFN和促炎性细胞因子信号传导途径均对MyXV感染响应；并专门确定哪些细胞传感器/换能器成分在转化的人髓样细胞中会功能失调。 AIM 2。创建含PYD的蛋白M013的工程变体，对炎症体和NF-κB信号传导途径进行不同的调节，并在体内caspase 1-依赖性大鼠移植血管疾病模型中测试这些M013构建体，并将仅与ASS-4 FAINS ASS-4 FATICTATION INTER pATICTATION cONTISTINAL ORS-NF-by-nf-bef-by-by-nf-by-nf-nf-be。 NF-κB1并利用这些突变体构建重组myxvs，以测试上述髓样细胞的作用。我们还将测试经过设计的M013变体，允许在大鼠同种异体移植血管疾病（TVD）模型中细胞内穿透模型，该模型已被炎症组介导的caspase 1的激活所调节，目标3。阐明宿主炎症和NF-κB信号的损害造成的孔隙率的损害的作用。炎症和 MYXV诱导的抗肿瘤反应中的NF-κB成分（IE ASC-1，caspase-1，NLRP3和NF-κB1）在NSG免疫症状的小鼠和C57BL/6型内的NSG免疫性小鼠中的人（例如THP-1）或鼠（例如RAW426.7）的鼠（例如RAW426.7）的清除率（例如THP-1）或鼠（例如RAW426.7）中的清除;研究使用M013KO与野生型MYXV的M013变体在选择性基因敲除小鼠中的作用，以及仅选择性地抑制炎症体或NF-κB途径的M013表达变体病毒。

项目成果

Recombinant Myxoma Virus-Derived Immune Modulator M-T7 Accelerates Cutaneous Wound Healing and Improves Tissue Remodeling.

• DOI: 10.3390/pharmaceutics12111003
• 发表时间: 2020-10-22
• 期刊: Pharmaceutics
• 影响因子: 5.4
• 作者: Yaron JR;Zhang L;Guo Q;Awo EA;Burgin M;Schutz LN;Zhang N;Kilbourne J;Daggett-Vondras J;Lowe KM;Lucas AR
• 通讯作者: Lucas AR

Targeting gallbladder cancer: oncolytic virotherapy with myxoma virus is enhanced by rapamycin in vitro and further improved by hyaluronan in vivo.

• DOI: 10.1186/1476-4598-13-82
• 发表时间: 2014-04-13
• 期刊: Molecular cancer
• 影响因子: 37.3
• 作者: Weng M;Gong W;Ma M;Chu B;Qin Y;Zhang M;Lun X;McFadden G;Forsyth P;Yang Y;Quan Z
• 通讯作者: Quan Z