CN113577242B - Nlrp12在制备治疗冠状病毒感染或相关的炎症性疾病的药物中的应用 - Google Patents
Nlrp12在制备治疗冠状病毒感染或相关的炎症性疾病的药物中的应用 Download PDFInfo
- Publication number
- CN113577242B CN113577242B CN202110423817.XA CN202110423817A CN113577242B CN 113577242 B CN113577242 B CN 113577242B CN 202110423817 A CN202110423817 A CN 202110423817A CN 113577242 B CN113577242 B CN 113577242B
- Authority
- CN
- China
- Prior art keywords
- nlrp12
- inflammatory
- coronavirus
- expression
- infection
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 208000001528 Coronaviridae Infections Diseases 0.000 title claims abstract description 17
- 208000027866 inflammatory disease Diseases 0.000 title claims abstract description 12
- 239000003814 drug Substances 0.000 title claims abstract description 7
- 102100039240 NACHT, LRR and PYD domains-containing protein 12 Human genes 0.000 title claims description 85
- 101000962345 Homo sapiens NACHT, LRR and PYD domains-containing protein 12 Proteins 0.000 title claims description 84
- 241000711573 Coronaviridae Species 0.000 claims abstract description 21
- 230000014509 gene expression Effects 0.000 claims description 29
- 108020004459 Small interfering RNA Proteins 0.000 claims description 14
- 101100025714 Homo sapiens NLRP12 gene Proteins 0.000 claims description 12
- 101150104437 NLRP12 gene Proteins 0.000 claims description 12
- 239000004055 small Interfering RNA Substances 0.000 claims description 11
- 102000053642 Catalytic RNA Human genes 0.000 claims description 4
- 108090000994 Catalytic RNA Proteins 0.000 claims description 4
- 108091027967 Small hairpin RNA Proteins 0.000 claims description 4
- 239000000074 antisense oligonucleotide Substances 0.000 claims description 4
- 238000012230 antisense oligonucleotides Methods 0.000 claims description 4
- 230000002222 downregulating effect Effects 0.000 claims description 4
- 108091092562 ribozyme Proteins 0.000 claims description 4
- 229940121649 protein inhibitor Drugs 0.000 claims description 2
- 239000012268 protein inhibitor Substances 0.000 claims description 2
- 108091034117 Oligonucleotide Proteins 0.000 claims 2
- 230000002068 genetic effect Effects 0.000 claims 1
- 230000002757 inflammatory effect Effects 0.000 abstract description 49
- 102000004127 Cytokines Human genes 0.000 abstract description 21
- 108090000695 Cytokines Proteins 0.000 abstract description 21
- 210000004072 lung Anatomy 0.000 abstract description 18
- 230000028327 secretion Effects 0.000 abstract description 7
- 230000003612 virological effect Effects 0.000 abstract description 5
- 230000007170 pathology Effects 0.000 abstract 1
- 230000008685 targeting Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 32
- 102000004889 Interleukin-6 Human genes 0.000 description 17
- 108090001005 Interleukin-6 Proteins 0.000 description 17
- 241000699670 Mus sp. Species 0.000 description 17
- 241001112090 Pseudovirus Species 0.000 description 17
- 238000001514 detection method Methods 0.000 description 16
- 208000015181 infectious disease Diseases 0.000 description 15
- 230000002829 reductive effect Effects 0.000 description 15
- 230000002401 inhibitory effect Effects 0.000 description 14
- 230000002018 overexpression Effects 0.000 description 14
- 230000002685 pulmonary effect Effects 0.000 description 14
- 108090000623 proteins and genes Proteins 0.000 description 11
- 210000001519 tissue Anatomy 0.000 description 11
- 241000699666 Mus <mouse, genus> Species 0.000 description 10
- 230000004913 activation Effects 0.000 description 10
- 210000002966 serum Anatomy 0.000 description 10
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 9
- 102100020881 Interleukin-1 alpha Human genes 0.000 description 9
- 230000001965 increasing effect Effects 0.000 description 9
- 230000035897 transcription Effects 0.000 description 9
- 238000013518 transcription Methods 0.000 description 9
- 101710085938 Matrix protein Proteins 0.000 description 8
- 101710127721 Membrane protein Proteins 0.000 description 8
- 230000030279 gene silencing Effects 0.000 description 8
- 230000008595 infiltration Effects 0.000 description 8
- 238000001764 infiltration Methods 0.000 description 8
- 108700002673 Coronavirus M Proteins Proteins 0.000 description 7
- 201000003176 Severe Acute Respiratory Syndrome Diseases 0.000 description 7
- 230000006870 function Effects 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- 108020003175 receptors Proteins 0.000 description 7
- 102000005962 receptors Human genes 0.000 description 7
- 230000004083 survival effect Effects 0.000 description 7
- 230000006378 damage Effects 0.000 description 6
- 230000003247 decreasing effect Effects 0.000 description 6
- 230000003993 interaction Effects 0.000 description 6
- 230000037361 pathway Effects 0.000 description 6
- 239000013612 plasmid Substances 0.000 description 6
- 230000001105 regulatory effect Effects 0.000 description 6
- 238000001890 transfection Methods 0.000 description 6
- 230000009385 viral infection Effects 0.000 description 6
- 102100022691 NACHT, LRR and PYD domains-containing protein 3 Human genes 0.000 description 5
- 206010035664 Pneumonia Diseases 0.000 description 5
- 108010001946 Pyrin Domain-Containing 3 Protein NLR Family Proteins 0.000 description 5
- 208000036142 Viral infection Diseases 0.000 description 5
- 241000700605 Viruses Species 0.000 description 5
- 239000013613 expression plasmid Substances 0.000 description 5
- 210000002540 macrophage Anatomy 0.000 description 5
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 5
- 230000019491 signal transduction Effects 0.000 description 5
- 208000024891 symptom Diseases 0.000 description 5
- 108700003471 Coronavirus 3C Proteases Proteins 0.000 description 4
- 101000979565 Homo sapiens Protein NLRC5 Proteins 0.000 description 4
- 101001074035 Homo sapiens Zinc finger protein GLI2 Proteins 0.000 description 4
- 108010006444 Leucine-Rich Repeat Proteins Proteins 0.000 description 4
- 102100023432 Protein NLRC5 Human genes 0.000 description 4
- 102000005583 Pyrin Human genes 0.000 description 4
- 108010059278 Pyrin Proteins 0.000 description 4
- 238000011529 RT qPCR Methods 0.000 description 4
- 208000027418 Wounds and injury Diseases 0.000 description 4
- 102100035558 Zinc finger protein GLI2 Human genes 0.000 description 4
- 230000028709 inflammatory response Effects 0.000 description 4
- 208000014674 injury Diseases 0.000 description 4
- 238000011813 knockout mouse model Methods 0.000 description 4
- 210000004901 leucine-rich repeat Anatomy 0.000 description 4
- 239000013642 negative control Substances 0.000 description 4
- 239000002773 nucleotide Substances 0.000 description 4
- 125000003729 nucleotide group Chemical group 0.000 description 4
- 230000001737 promoting effect Effects 0.000 description 4
- 108090000426 Caspase-1 Proteins 0.000 description 3
- 241000282412 Homo Species 0.000 description 3
- 206010037660 Pyrexia Diseases 0.000 description 3
- 101000953880 Severe acute respiratory syndrome coronavirus 2 Membrane protein Proteins 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 210000004969 inflammatory cell Anatomy 0.000 description 3
- 230000001404 mediated effect Effects 0.000 description 3
- 102000007863 pattern recognition receptors Human genes 0.000 description 3
- 108010089193 pattern recognition receptors Proteins 0.000 description 3
- 230000000770 proinflammatory effect Effects 0.000 description 3
- 239000003531 protein hydrolysate Substances 0.000 description 3
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 2
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 description 2
- 108020000948 Antisense Oligonucleotides Proteins 0.000 description 2
- 208000035143 Bacterial infection Diseases 0.000 description 2
- 102000021350 Caspase recruitment domains Human genes 0.000 description 2
- 108091011189 Caspase recruitment domains Proteins 0.000 description 2
- 102100035904 Caspase-1 Human genes 0.000 description 2
- 206010011224 Cough Diseases 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- -1 G-SCF Proteins 0.000 description 2
- 102000014150 Interferons Human genes 0.000 description 2
- 108010050904 Interferons Proteins 0.000 description 2
- 102000003777 Interleukin-1 beta Human genes 0.000 description 2
- 108090000193 Interleukin-1 beta Proteins 0.000 description 2
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 2
- 241000127282 Middle East respiratory syndrome-related coronavirus Species 0.000 description 2
- 101710084317 NACHT, LRR and PYD domains-containing protein 12 Proteins 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 108091007494 Nucleic acid- binding domains Proteins 0.000 description 2
- 241000315672 SARS coronavirus Species 0.000 description 2
- 108010065667 Viral Matrix Proteins Proteins 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000010100 anticoagulation Effects 0.000 description 2
- 230000006907 apoptotic process Effects 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 208000022362 bacterial infectious disease Diseases 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 108700010904 coronavirus proteins Proteins 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 230000007783 downstream signaling Effects 0.000 description 2
- 239000012636 effector Substances 0.000 description 2
- 210000002919 epithelial cell Anatomy 0.000 description 2
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 2
- 238000011065 in-situ storage Methods 0.000 description 2
- 229940047124 interferons Drugs 0.000 description 2
- 201000005202 lung cancer Diseases 0.000 description 2
- 210000005265 lung cell Anatomy 0.000 description 2
- 208000020816 lung neoplasm Diseases 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 230000008807 pathological lesion Effects 0.000 description 2
- 210000005259 peripheral blood Anatomy 0.000 description 2
- 239000011886 peripheral blood Substances 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 208000023504 respiratory system disease Diseases 0.000 description 2
- 230000000451 tissue damage Effects 0.000 description 2
- 231100000827 tissue damage Toxicity 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 241000228245 Aspergillus niger Species 0.000 description 1
- 102100021943 C-C motif chemokine 2 Human genes 0.000 description 1
- 102100025248 C-X-C motif chemokine 10 Human genes 0.000 description 1
- 101710098275 C-X-C motif chemokine 10 Proteins 0.000 description 1
- 108700012434 CCL3 Proteins 0.000 description 1
- 208000025721 COVID-19 Diseases 0.000 description 1
- 241001678559 COVID-19 virus Species 0.000 description 1
- 102000000013 Chemokine CCL3 Human genes 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 241000701022 Cytomegalovirus Species 0.000 description 1
- 208000000059 Dyspnea Diseases 0.000 description 1
- 206010013975 Dyspnoeas Diseases 0.000 description 1
- 102100038513 E3 ubiquitin-protein ligase ARIH2 Human genes 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 108091062183 EsiRNA Proteins 0.000 description 1
- 102100037388 Gasdermin-D Human genes 0.000 description 1
- 102100028539 Guanylate-binding protein 5 Human genes 0.000 description 1
- 101710110795 Guanylate-binding protein 5 Proteins 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- 101000808888 Homo sapiens E3 ubiquitin-protein ligase ARIH2 Proteins 0.000 description 1
- 101001026262 Homo sapiens Gasdermin-D Proteins 0.000 description 1
- 101000852483 Homo sapiens Interleukin-1 receptor-associated kinase 1 Proteins 0.000 description 1
- 101000979575 Homo sapiens NLR family CARD domain-containing protein 3 Proteins 0.000 description 1
- 101000979572 Homo sapiens NLR family CARD domain-containing protein 4 Proteins 0.000 description 1
- 101001125032 Homo sapiens Nucleotide-binding oligomerization domain-containing protein 1 Proteins 0.000 description 1
- 101001125026 Homo sapiens Nucleotide-binding oligomerization domain-containing protein 2 Proteins 0.000 description 1
- 101000588545 Homo sapiens Serine/threonine-protein kinase Nek7 Proteins 0.000 description 1
- 241000711467 Human coronavirus 229E Species 0.000 description 1
- 241001109669 Human coronavirus HKU1 Species 0.000 description 1
- 241000482741 Human coronavirus NL63 Species 0.000 description 1
- 241001428935 Human coronavirus OC43 Species 0.000 description 1
- 102000002227 Interferon Type I Human genes 0.000 description 1
- 108010014726 Interferon Type I Proteins 0.000 description 1
- 102000000589 Interleukin-1 Human genes 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- 102100036342 Interleukin-1 receptor-associated kinase 1 Human genes 0.000 description 1
- 108010082786 Interleukin-1alpha Proteins 0.000 description 1
- 208000004852 Lung Injury Diseases 0.000 description 1
- 208000019693 Lung disease Diseases 0.000 description 1
- 102000043136 MAP kinase family Human genes 0.000 description 1
- 108091054455 MAP kinase family Proteins 0.000 description 1
- 101710091439 Major capsid protein 1 Proteins 0.000 description 1
- 208000025370 Middle East respiratory syndrome Diseases 0.000 description 1
- 102100024193 Mitogen-activated protein kinase 1 Human genes 0.000 description 1
- 241000711408 Murine respirovirus Species 0.000 description 1
- 101100025715 Mus musculus Nlrp12 gene Proteins 0.000 description 1
- 241000187479 Mycobacterium tuberculosis Species 0.000 description 1
- 108010084333 N-palmitoyl-S-(2,3-bis(palmitoyloxy)propyl)cysteinyl-seryl-lysyl-lysyl-lysyl-lysine Proteins 0.000 description 1
- 102000012064 NLR Proteins Human genes 0.000 description 1
- 102100023382 NLR family CARD domain-containing protein 3 Human genes 0.000 description 1
- 102100023435 NLR family CARD domain-containing protein 4 Human genes 0.000 description 1
- 101150042416 Nlrp1b gene Proteins 0.000 description 1
- 101710141454 Nucleoprotein Proteins 0.000 description 1
- 102100029424 Nucleotide-binding oligomerization domain-containing protein 1 Human genes 0.000 description 1
- 102100029441 Nucleotide-binding oligomerization domain-containing protein 2 Human genes 0.000 description 1
- 206010068319 Oropharyngeal pain Diseases 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 201000007100 Pharyngitis Diseases 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 108010009341 Protein Serine-Threonine Kinases Proteins 0.000 description 1
- 102000009516 Protein Serine-Threonine Kinases Human genes 0.000 description 1
- 240000000191 Prunus tomentosa Species 0.000 description 1
- 239000013614 RNA sample Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 208000036071 Rhinorrhea Diseases 0.000 description 1
- 206010039101 Rhinorrhoea Diseases 0.000 description 1
- 108091005774 SARS-CoV-2 proteins Proteins 0.000 description 1
- 102100031400 Serine/threonine-protein kinase Nek7 Human genes 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 101710196623 Stimulator of interferon genes protein Proteins 0.000 description 1
- 101710172711 Structural protein Proteins 0.000 description 1
- 206010051379 Systemic Inflammatory Response Syndrome Diseases 0.000 description 1
- 206010069363 Traumatic lung injury Diseases 0.000 description 1
- 108010087302 Viral Structural Proteins Proteins 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 230000033289 adaptive immune response Effects 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000001640 apoptogenic effect Effects 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 206010006451 bronchitis Diseases 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000014564 chemokine production Effects 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 238000000749 co-immunoprecipitation Methods 0.000 description 1
- 230000008045 co-localization Effects 0.000 description 1
- 239000000571 coke Substances 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000016396 cytokine production Effects 0.000 description 1
- 206010052015 cytokine release syndrome Diseases 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 230000005860 defense response to virus Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 241001493065 dsRNA viruses Species 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- 208000019622 heart disease Diseases 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 210000003917 human chromosome Anatomy 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000010166 immunofluorescence Methods 0.000 description 1
- 238000011532 immunohistochemical staining Methods 0.000 description 1
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 230000004957 immunoregulator effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000006749 inflammatory damage Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 230000015788 innate immune response Effects 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 231100000515 lung injury Toxicity 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000001616 monocyte Anatomy 0.000 description 1
- 210000005087 mononuclear cell Anatomy 0.000 description 1
- 108091005763 multidomain proteins Proteins 0.000 description 1
- 230000008383 multiple organ dysfunction Effects 0.000 description 1
- 239000007923 nasal drop Substances 0.000 description 1
- 229940100662 nasal drops Drugs 0.000 description 1
- 201000009240 nasopharyngitis Diseases 0.000 description 1
- 230000004719 natural immunity Effects 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 238000006384 oligomerization reaction Methods 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 230000031339 positive regulation of inflammatory response Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000007115 recruitment Effects 0.000 description 1
- 102000037983 regulatory factors Human genes 0.000 description 1
- 108091008025 regulatory factors Proteins 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 208000013220 shortness of breath Diseases 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- 241000701447 unidentified baculovirus Species 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 230000029812 viral genome replication Effects 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/177—Receptors; Cell surface antigens; Cell surface determinants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Virology (AREA)
- Analytical Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Immunology (AREA)
- Wood Science & Technology (AREA)
- Animal Behavior & Ethology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Genetics & Genomics (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Oncology (AREA)
- Communicable Diseases (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Gastroenterology & Hepatology (AREA)
- Cell Biology (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Epidemiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本发明属于生物技术和医学技术领域，本发明通过靶向NLRP12，有效抑制炎性细胞因子分泌，降低病毒载量，改善肺部病理。通过本申请公开的方法，可实现对冠状病毒引起的炎症性疾病的控制，为冠状病毒感染的治疗提供靶点。
Description
技术领域
本发明涉及生物技术和医学领域，更具体地，涉及核苷酸结合结构域和富含亮氨酸重复 结构域的受体12(nucleotide-binding domain leucine-rich repeat-containing receptors 12， NLRP12)在制备治疗冠状病毒感染或相关的炎症性疾病的药物中的应用。
背景技术
冠状病毒是广泛存在于自然界，具囊膜(envelope)、基因组为线性单股正链的RNA病 毒。目前已经发现7种可感染人的冠状病毒。其中常见的229E、NL63、OC43和HKU1型通 常会引起轻度或中度的上呼吸道疾病，如感冒。症状主要包括流鼻涕、头痛、咳嗽、咽喉痛、 发热等，有时会引起肺炎或支气管炎等下呼吸道疾病；在心肺疾病患者、免疫力低下人群、 婴儿和老年人中较为常见。MERS-CoV，SARS-CoV常引起较为严重症状。MERS症状通常 包括发热、咳嗽和呼吸急促，甚至发展为肺炎，病死率约为34.4％。SARS症状通常包括发热、 畏寒和身体疼痛，甚至发展为肺炎，病死率约为9.6％。此外，SARS-CoV-2病毒也能引起严 重症状，约有5％的患者恶化为严重的肺损伤甚至多器官功能障碍，综合死亡率约为2％。
冠状病毒入侵机体后，会被宿主细胞中的多种模式识别受体(PRRs)所识别，进而引发 天然免疫信号通路的活化，产生一系列炎性细胞因子。研究发现，COVID-19患者的炎性细 胞因子TNF-α、IL-1β、IL-6、G-SCF、IP-10、MCP1和MIP1α水平显著升高。已有研究发现，细胞因子风暴引发的过度炎症反应在COVID-19的免疫致病机理中起重要作用，约73.1％的新冠病人死于全身性炎症反应综合征。因此抑制过度的炎症反应是控制新冠感染的重要手段。
核苷酸结合寡聚化结构域样受体(Nod-like receptors，NLRs)家族蛋白是一类广泛存在 于细胞质内的PRRs，通过调控机体的炎症信号通路，参与天然免疫应答。NLRP12是NLRs 家族重要的成员，能够靶向不同的炎症信号通路发挥调控作用。NLRP12(NLR familyPYRIN domain containing 12)，又称PYPAF7(PYRIN-containing Apaf1-like protein7)，是NLRP亚 家族中的一员，其编码基因位于人19号染色体上。与大多数NLRP受体一样，NLRP12由 三大结构域组成，分别为N端的PYRIN结构域、中间的NACHT结构域及C端的LRRs结构域。体外研究发现，NLRP12具有较为广泛的调控作用。一方面，NLRP12能够负调控 经典/非经典NF-κB信号通路、抑制促炎因子和趋化因子产生。NLRP12通过与IRAK1相互 作用，并阻碍其磷酸化，可有效抑制Pam3Cys(TLR1/2激活剂)或LPS介导的经典NF-κB 通路的活化；同时，NLRP12还能与NIK相互作用，通过蛋白酶途径水解NIK，从而显著 抑制CD40介导的非经典NF-κB通路的活化。Dr.Ting实验室在结核分枝杆菌感染的 THP-1单核细胞系中，下调NLRP12表达后，发现经典NF-κB通路的活化明显增强，导致 炎性因子IL-6大量产生。随后，Dr.Kanneganti实验室在沙门氏菌感染的巨噬细胞中，发现 NLRP12不仅具有抑制经典NF-κB通路活化的功能，还能够明显抑制MAPKs/ERK通路活 化，从而显著减少IL-6、TNF-α和NO的产生。此外NLRP12还能组装炎症小体的组装和 中性粒细胞的招募和激活以发挥促炎作用。但是NLRP12在抗冠状病毒感染领域内的功能还 没有确切报道。
综上所述，在抗冠状病毒感染的天然免疫领域内，目前迫切需要开发能够增强机体抗病 毒反应能力，防止炎症反应过度活化造成免疫炎症损伤的免疫调节分子。
发明内容
本发明为了克服现有技术中存在的上述问题，提供了核苷酸结合结构域和富含亮氨酸重 复结构域的受体12(nucleotide-binding domain leucine-rich repeat-containing receptors 12， NLRP12)在制备治疗冠状病毒感染或相关的炎症性疾病的药物中的应用。
本发明的目的通过以下技术方案实现：
NLRP12在制备抗冠状病毒感染和/或冠状病毒感染相关的炎症性疾病的功能产品中的应 用。
优选的，所述功能产品具有下调NLRP12基因的表达、转录、或者其表达产物的功能。
优选的，所述冠状病毒包括但不仅限于新型冠状病毒、人冠状病毒229E、人冠状病毒 OC43、人冠状病毒NL63、人冠状病毒HKU1、SARS病毒、MERS病毒及小鼠冠状病毒。
更优选的，所述冠状病毒选自新型冠状病毒和小鼠冠状病毒。
优选的，所述功能产品能够抑制冠状病毒感染引起的炎性因子的分泌。
更优选的，所述炎性因子选自IL-1α，IL-1β和IL-6。
本发明发现，在肺上皮细胞系A549和巨噬细胞系THP-1中过表达NLRP12能够促进新 冠病毒M蛋白假病毒引起的IL-1α，IL-1β和IL-6分泌；沉默NLRP12能够抑制新冠病毒M蛋白假病毒引起的IL-1α，IL-1β和IL-6分泌。在小鼠中敲除NLRP12，然后再用小鼠冠状病毒MHV-A59进行肺部感染，敲除小鼠和未敲除小鼠相比，发生了以下变化：(1)生存率增 加；(2)肺部病毒载量降低；(3)肺部炎性浸润和组织损伤减少；(4)血清炎性细胞因子水 平降低。
因此，在实际应用中，可以通过抑制NLRP12基因及表达产物，以达到抑制冠状病毒， 减轻炎症损伤的目的。
更优选的，NLRP12基因的表达产物是指NLRP12基因在各阶段中的各种形式的分子， 例如但不限于NLRP12基因在扩增、复制、转录、剪接、加工、翻译、修饰过程中所产生的分子，例如cDNA、mRNA、前体蛋白、成熟的蛋白、及其片段等。
作为一种优选的实施方式，本发明上述功能产品包括：NLRP12蛋白抑制剂、NLRP12基因缺陷或者沉默的免疫相关细胞、其分化细胞或者基因重组构建体中的一种或者多种。
更优选的，所述功能产品包括以下任意一种：
(i)以NLRP12转录本为靶序列，且能够激活NLRP12基因表达产物的表达或基因转录 的反义寡核苷酸、siRNA、dsRNA、核酶、esiRNA、shRNA等；
(ii)能表达或者形成(i)中所述反义寡核苷酸、siRNA、dsRNA、核酶、esiRNA、shRNA的构建体等；
(iii)含有NLRP12互补序列，且能够在转入体内后形成抑制NLRP12基因表达产物的 表达或基因转录的干扰分子的构建体；
(iii)抑制或者敲除了NLRP12基因序列后的免疫相关细胞、其分化细胞或构建体。
本发明研究发现通过沉默或敲除NLRP12能够明显抑制新冠病毒蛋白和MHV引起的炎 性损伤，提示NLRP12具有促进冠状病毒感染的效果，有望成为治疗冠状病毒感染的一个靶 点。
优选的，上述治疗包括至少一种情况：抑制受试对象体内病毒复制、提高受试对象存活 率、抑制受试对象体内炎性因子IL-1α，IL-1β和IL-6的水平。
与现有技术相比，本发明具有以下有益效果：
本发明研究发现通过沉默或敲除NLRP12能够明显抑制新冠病毒蛋白和MHV引起的炎 性损伤，提示NLRP12具有促进冠状病毒感染的效果，有望成为治疗新冠病毒肺炎的一个靶 点。在肺上皮细胞系A549和巨噬细胞系THP-1中过表达NLRP12能够促进新冠病毒M蛋白假病毒引起的IL-1α，IL-1β和IL-6分泌；沉默NLRP12能够抑制新冠病毒M蛋白假病毒引 起的IL-1α，IL-1β和IL-6分泌。在小鼠中敲除NLRP12，然后再用小鼠冠状病毒MHV-A59) 进行肺部感染，敲除小鼠和未敲除小鼠相比，发生了以下变化：(1)生存率增加；(2)肺部 病毒载量降低；(3)肺部炎性浸润和组织损伤减少；(4)血清炎性细胞因子水平降低。因此， 在实际应用中，可以通过抑制NLRP12基因及表达产物，以达到抑制冠状病毒，减轻炎症损 伤的目的。从本质上证明了NLRP12在治疗冠状病毒感染及冠状病毒感染的引发的炎症性疾病中具有广泛的应用背景。
附图说明
图1显示新冠感染后，人外周血单个核细胞和肺组织中NLRP12表达上调；图中*代表 P<0.05、**代表P<0.01、***代P<0.001；图1A为NLRP12在新冠病人和健康人外周血单个核细胞中的RNA水平；图1B为流式检测新冠病人和健康人外周血单个核细胞中不同细胞亚群上NLRP12的表达水平；图1C为新冠病人和健康人外周血单个核细胞中NLRP12表达 的平均荧光强度；图1D为免疫组化染色检测肺部原位的NLRP12表达，取癌症患者远端癌 旁组织作为阴性对照，取肺癌组织为阳性对照；
图2显示NLRP12能和新冠病毒膜蛋白发生相互作用，且依赖于其LRRs结构域；图2A为在293T细胞中外源转入NLRP12和新冠病毒的四个结构蛋白的免疫共沉淀实验；图2B为在293T细胞外源转入NLRP12和新冠病毒膜蛋白的共定位；图2C为NLRP12不同结构域与 新冠M蛋白的相互作用；
图3A显示使用NLRP12小干扰RNA沉默内源性NLRP12后，SARS-COV-2的M蛋白 假病毒刺激THP1细胞的炎性细胞因子IL-1α，IL-1β和IL-6RNA水平降低；图3B显示使 用NLRP12过表达质粒过表达NLRP12后，SARS-COV-2的M蛋白假病毒刺激THP1细胞的 炎性细胞因子IL-1α，IL-1β和IL-6RNA水平上升；图中*代表P<0.05、**代表P<0.01、*** 代P<0.001；
图4A显示使用NLRP12小干扰RNA沉默内源性NLRP12后，SARS-COV-2的M蛋白 假病毒刺激A549细胞的炎性细胞因子IL-1α，IL-1β和IL-6RNA水平降低；图4B显示使 用NLRP12过表达质粒过表达NLRP12后，SARS-COV-2的M蛋白假病毒刺激A549细胞的 炎性细胞因子IL-1α，IL-1β和IL-6RNA水平升高；图中*代表P<0.05、**代表P<0.01、*** 代P<0.001；
图5显示敲除小鼠NLRP12后，SARS-COV-2的M蛋白假病毒感染后小鼠肺部炎性浸润减少，肺部细胞因子转录水平降低，血清炎性细胞因子表达降低；其中图5A为肺部的M.cherry表达情况；图5B为小鼠的肺部病理损伤情况；图5C为肺部巨噬细胞占总细胞比例代表图； 图5D为肺部巨噬细胞占总细胞比例统计图；图5E-5F为肺部IL-1α、IL-1β和IL-6RNA水平；图5G为血清TNF、IL-1β和IL-6水平；图中Mock代表未过表达NLRP12的VSV-G空 载病毒，VSV-G-M代表过表达新冠膜蛋白的VSV-G病毒，Wt为野生小鼠，NLRP12-/-代表 敲除NLRP12小鼠；
图6显示敲除小鼠NLRP12后，小鼠冠状病毒MHV-A59感染后小鼠生存率增加，肺部病毒载量降低，肺部炎性浸润减少，肺部细胞因子转录水平降低，血清炎性细胞因子表达降低；图6A为小鼠生存率统计图；图6B为小鼠的肺部病理损伤情况；图6C为CT检测肺部 炎症情况；图6D为小鼠肺部病毒N蛋白RNA水平图；图6E为肺部IL-1α、IL-1β和IL-6RNA 水平；图6F为血清TNF、IL-1β和IL-6水平，图中MHV代表小鼠冠状病毒MHV-A59滴鼻处理，并以PBS为对照组。
具体实施方式
下面对本发明的具体实施方式作进一步说明。在此需要说明的是，对于这些实施方式的 说明用于帮助理解本发明，但并不构成对本发明的限定。此外，下面所描述的本发明各个实 施方式中所涉及的技术特征只要彼此之间未构成冲突就可以相互组合。
下述实验例中所使用的试验方法如无特殊说明，均为常规方法；所使用的材料、试剂等， 如无特殊说明，为可从商业途径得到的试剂和材料。
实施例1新冠感染后，NLRP12表达水平上调
1.1、检测样本收集：
收集新冠病人和健康人外周血单个核细胞的总RNA样本。
1.2、检测方法：
EDTA抗凝管收集新冠病人和体检健康人的外周血5mL，用生理盐水将人外周抗凝血按 1:2或1:3比例稀释，轻轻混匀。将稀释后的血液按1:1比例缓慢加入到已有淋巴细胞分离液 的上层，室温水平离心(1800rpm，30min)，使血细胞分层。吸取中间白膜层(单个核细胞 层)，加入生理盐水，轻轻混匀，4℃离心，1500rpm，10min。收集细胞沉淀。一部分用于流式检测，一部分加入500uL Trizol，对总RNA提取，Quantitative Real-time PCR(Q-RT-PCR) 检测。免疫组化取癌症患者远端癌旁组织作为阴性对照，取肺癌组织为阳性对照。
1.3、实验结果：
结果如图1所示，新型冠状病毒感染后，NLRP12基因水平表达上升，且在外周血中的 单核细胞上表达增加。此外，在肺组织原位NLRP12也表达上调；提示NLRP12在冠状病毒感染中发挥重要作用。
实施例2 NLRP12的LRRs结构域和新冠病毒膜蛋白发生直接相互作用
2.1检测样本收集：
收集转入NLRP12质粒和病毒结构蛋白的293T细胞蛋白裂解液。收集转入NLRP12分段克隆质粒和病毒M蛋白的293T细胞蛋白裂解液。
2.2检测方法：
蛋白裂解液加入HA凝胶珠孵育过夜并洗涤，Western Blot检测相互作用。免疫荧光用 HA和Flag的抗体孵育转入NLRP12质粒和病毒M蛋白的293T细胞，共聚焦观察。
2.3实验结果：
如图2所示，NLRP12能够与M蛋白相互作用，且这种相互作用依赖于LRRs结构域。
实施例3 SARS-COV-2的M蛋白假病毒刺激THP1细胞的细胞因子转录水平测定实验
3.1、检测样本的收集：
收集NLRP12沉默或过表达后，VSV-G-M感染的THP-1细胞的总RNA。
3.2、检测方法，具体包括如下步骤：
(1)THP-1细胞按照5×105个/ml的密度接种12孔板并用50ng//mL PMA刺激，培养过 夜。
(2)第二天弃掉培养液，用1×PBS洗两遍。购买NLRP12的小干扰RNA，或用NLRP12的过表达质粒，转染THP1细胞，然后病毒以MOI＝1感染细胞，培养6h，12h和24h。
(3)收集细胞总RNA，Q-RT-PCR检测IL-1α，IL-1β和IL-6的转录水平。
3.3、实验结果：
结果如图3所示，0h为不感染组，siNC组为转染阴性对照小干扰RNA，siNLRP12组为转染NLRP12小干扰RNA；Vector组为转染对照质粒组，NLRP12组为转染NLRP12过表达 质粒组；由结果可知，转染NLRP12小干扰RNA后，明显抑制了M假病毒感染产生的炎性 因子IL1α、IL1β、IL6表达水平，说明抑制NLRP12有效抑制炎性因子的产生，减轻炎症性 疾病；反之，过表达NLRP12后，明显促进了M假病毒感染产生的炎性因子IL1α、IL1β、IL6 表达水平，说明过表达NLRP12有效促进炎性因子的产生，加重炎症性疾病。
实施例4 SARS-COV-2的M蛋白假病毒刺激A549细胞的细胞因子转录水平测定实验
3.1、检测样本的收集：
收集NLRP12沉默或过表达后，SARS-COV-2-M蛋白假病毒感染的A549细胞的总RNA。
3.2、检测方法，具体包括如下步骤：
(1)A549细胞按照2×105个/ml的密度接种12孔板，并用50ng//mL PMA刺激，培养过夜。
(2)第二天弃掉培养液，用1×PBS洗两遍。购买NLRP12的小干扰RNA，或用NLRP12的过表达质粒，转染A549细胞，然后病毒以MOI＝1感染细胞，培养6h，12h和24h。
(3)收集细胞总RNA，Q-RT-PCR检测IL-1α，IL-1β和IL-6的转录水平。
3.3、实验结果：
结果如图4所示，0h为不感染组，siNC组为转染阴性对照小干扰RNA，siNLRP12组为转染NLRP12小干扰RNA；Vector组为转染对照质粒组，NLRP12组为转染NLRP12过表达 质粒组；由结果可知，过表达NLRP12后，明显促进了M假病毒感染产生的炎性因子IL1α、 IL1β、IL6表达水平，说明过表达NLRP12有效促进炎性因子的产生，加重炎症性疾病；反之转染NLRP12小干扰RNA后，明显抑制了M假病毒感染产生的炎性因子IL1α、IL1β、IL6 表达水平，说明抑制NLRP12有效抑制炎性因子的产生，减轻炎症性疾病。
实施例5小鼠敲除NLRP12后，SARS-COV-2的M蛋白假病毒感染后检测肺部炎性浸润和 病理损伤
通过气道内注射感染SARS-COV-2的M蛋白假病毒(每只5×106PFU)，72小时后，取肺脏组织做HE染色，取肺脏总细胞做流式检测和q-PCR分析，取血清做ELISA分析。
结果如图5所示，与野生型小鼠相比，NLRP12敲除小鼠肺组织内的炎症浸润和实变程 度明显减轻，炎性细胞浸润减少，肺组织中细胞因子转录水平降低，血清中的炎性细胞因子 含量降低。以上的结果表明，小鼠体内敲除NLRP12的表达后，小鼠对病毒感染的抵抗能力 增强。
实施例6小鼠敲除NLRP12后，MHV-A59感染后检测肺部炎性浸润和病理损伤
通过滴鼻感染MHV-A59(每只4×105PFU)，观察一周内的生存率。另取一组小鼠，通过滴鼻感染MHV-A59(每只4×105PFU)，取肺脏组织做HE染色，取肺脏总细胞做流式检 测和q-PCR分析，取血清做ELISA分析。
结果如图6所示，与野生型小鼠相比，NLRP12敲除小鼠生存率增加，病毒载量降低，肺组织内的炎症浸润和实变程度明显减轻，炎性细胞浸润减少，血清炎性细胞因子水平降低。 以上的结果表明，小鼠体内敲除NLRP12的表达后，小鼠对病毒感染的抵抗能力增强。
讨论：NLRs家族共有22个成员，均为具有三重结构的多结构域蛋白，即C端的LRRS结构域，中间的NACHT结构域及N端的效应结构域。三个结构域发挥的功能各不相同，其 中LRRs结构域的主要功能是介导蛋白与蛋白之间的相互作用，NACHT结构域能与核苷酸互 相结合并具有ATPase类似的功能，N端的效应结构域可以是pyrin结构域(PYD)、caspase 募集结构域(CARD)或杆状病毒凋亡蛋白重复结构域(BIR)，均与下游信号分子的连接及 下游信号通路的激活相关。以NLRP3为例，在LPS诱导下，NLRP3通过NOD和LRRs结构 域与NEK7结合，并与ASC一起形成炎症小体，正向调控炎症反应；在应对ATP及黑曲霉 素等致病菌的反应中，NLRP3通过PYRIN结构域和鸟苷酸结合蛋白5(GBP5)直接作用， 促进ASC的招募进而促进炎症小体的组装；此外，NLRP3的NACHT结构域还能与ARIH2 相互作用，并接收ARIH2的负调控信号。
尽管具有相似的结构域，NLRs家族不同成员发挥的功能却并不相同。例如，在细菌感染 中，NLRP3、NLRC4、NLRP1B能够发生构象变化，并与凋亡相关斑点样蛋白(ASC)组装 和caspase-1形成炎症小体，促进Casepase-1前体的剪切，形成具有催化活性的蛋白酶Caspase-1，介导促炎细胞因子IL-1β和IL-18的激活，并介导GSDMD介导的细胞焦亡。除了这些典型的介导炎症小体组装和激活的NLRs，其他NLRs也可作为关键炎症信号通路的正 向调节因子。例如，NLRC1和NLRC2识别细菌成分后被激活，通过CARD-CARD相互作用 直接招募丝氨酸-苏氨酸激酶受体相互作用蛋白类相互作用激酶(RICK)。RICK与NFκB基 本调节因子结合(NEMO)促进IKKα和IKKβ的活化，导致IκB的降解和NFκB向核的释放和转运，促进炎性细胞因子的分泌。此外，在细菌感染中，一些NLRs还能够抑制机体的 炎症反应。例如病毒感染时，NLRC3负调控STING信号通路，抑制炎性细胞因子的产生。 NLRC5也在病毒感染中抑制炎性细胞因子和干扰素的产生。综上所述，尽管NLRs家族不同 成员具有相似的结构域，但是不同的NLRs分子在感染性疾病中发挥的作用并不相同。
除此之外，一些NLRs在不同的疾病模型中还发挥不同的免疫调控作用，例如，NLRC5 在宿主防御过程中作为先天和适应性免疫应答的调节因子的作用还存在争议。病毒感染后， NLRC5抑制炎性细胞因子和干扰素的产生。然而，在巨细胞病毒或仙台病毒感染期间，NLRC5 促进成纤维细胞和原代人类细胞中I型干扰素和炎性细胞因子的产生。
Claims (3)
1.功能产品在制备治疗冠状病毒感染相关的炎症性疾病的药物中的应用，所述功能产品具有下调NLRP12基因的表达的功能，所述冠状病毒为新型冠状病毒SARS-COV-2或小鼠冠状病毒MHV-A59。
2.根据权利要求1所述的应用，其特征在于，所述功能产品为NLRP12蛋白抑制剂或者基因重组构建体中的一种或者两种。
3.根据权利要求1所述的应用，其特征在于，所述功能产品为以下任意一种：
（i）以NLRP12转录本为靶序列，且能够下调NLRP12基因的表达的反义寡核苷酸、siRNA、核酶或shRNA；
（ii）能表达或者形成（i）中所述反义寡核苷酸、siRNA、核酶或shRNA的构建体。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110423817.XA CN113577242B (zh) | 2021-04-20 | 2021-04-20 | Nlrp12在制备治疗冠状病毒感染或相关的炎症性疾病的药物中的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110423817.XA CN113577242B (zh) | 2021-04-20 | 2021-04-20 | Nlrp12在制备治疗冠状病毒感染或相关的炎症性疾病的药物中的应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113577242A CN113577242A (zh) | 2021-11-02 |
| CN113577242B true CN113577242B (zh) | 2023-12-08 |
Family
ID=78243022
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110423817.XA Active CN113577242B (zh) | 2021-04-20 | 2021-04-20 | Nlrp12在制备治疗冠状病毒感染或相关的炎症性疾病的药物中的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113577242B (zh) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117388500A (zh) * | 2023-08-03 | 2024-01-12 | 暨南大学 | Trpv4作为药物靶点在冠状病毒感染抗炎药物中的应用 |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111879943A (zh) * | 2020-07-10 | 2020-11-03 | 中山大学附属第五医院 | Slamf7重组蛋白在制备治疗新冠肺炎药物中的应用 |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015036009A1 (en) * | 2013-09-10 | 2015-03-19 | Polichem S.A. | Pidotimod for use in the treatment of inflammation-associated diseases |
-
2021
- 2021-04-20 CN CN202110423817.XA patent/CN113577242B/zh active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111879943A (zh) * | 2020-07-10 | 2020-11-03 | 中山大学附属第五医院 | Slamf7重组蛋白在制备治疗新冠肺炎药物中的应用 |
Non-Patent Citations (1)
| Title |
|---|
| SARS-CoV-2 proteases PLpro and 3CLpro cleave IRF3 and critical modulators of inflammatory pathways (NLRP12 and TAB1): implications for disease presentation across species;Mehdi Moustaqil等;《Emerging Microbes & Infections》;第10卷;第178-195页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113577242A (zh) | 2021-11-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Nabe | Steroid-resistant asthma and neutrophils | |
| Louahed et al. | Interleukin-9 upregulates mucus expression in the airways | |
| Shim et al. | IL-13 induces mucin production by stimulating epidermal growth factor receptors and by activating neutrophils | |
| Wu et al. | Activation of NLRP3 inflammasome in alveolar macrophages contributes to mechanical stretch-induced lung inflammation and injury | |
| Kim et al. | Interleukin‐22 promotes osteoclastogenesis in rheumatoid arthritis through induction of RANKL in human synovial fibroblasts | |
| Hardarson et al. | Toll-like receptor 3 is an essential component of the innate stress response in virus-induced cardiac injury | |
| Mellett et al. | Orphan receptor IL-17RD tunes IL-17A signalling and is required for neutrophilia | |
| Darcan-Nicolaisen et al. | Small interfering RNA against transcription factor STAT6 inhibits allergic airway inflammation and hyperreactivity in mice | |
| Wang et al. | Basic fibroblast growth factor protects against influenza A virus-induced acute lung injury by recruiting neutrophils | |
| Haas et al. | Inhibition of angiogenesis by interleukin‐4 gene therapy in rat adjuvant‐induced arthritis | |
| Rosas-Taraco et al. | Intrapulmonary delivery of XCL1-targeting small interfering RNA in mice chronically infected with Mycobacterium tuberculosis | |
| Boukhvalova et al. | Activation of interferon response through toll-like receptor 3 impacts viral pathogenesis and pulmonary toll-like receptor expression during respiratory syncytial virus and influenza infections in the cotton rat Sigmodon hispidus model | |
| Wu et al. | Intranasal sirna targeting c-kit reduces airway inflammation in experimental allergic asthma | |
| Lee et al. | Phosphatase and tensin homolog deleted on chromosome 10 (PTEN) reduces vascular endothelial growth factor expression in allergen-induced airway inflammation | |
| Lee et al. | MicroRNA-21 inhibition suppresses alveolar M2 macrophages in an ovalbumin-induced allergic asthma mice model | |
| US20220195432A9 (en) | Methods for preventing and treating or diagnosing coronavirus infection | |
| CN113577242B (zh) | Nlrp12在制备治疗冠状病毒感染或相关的炎症性疾病的药物中的应用 | |
| US20110184045A1 (en) | Silencng and rig-i activation by dual function oligonucleotides | |
| Li et al. | Sesamin attenuates allergic airway inflammation through the suppression of nuclear factor‑kappa B activation | |
| Liu et al. | Impact of TREM-2 gene silencing on inflammatory response of endotoxin-induced acute lung injury in mice | |
| EP3524274A2 (en) | Managing the treatment of a cellular proliferative disorder using hom-1 expression | |
| Hu et al. | Human umbilical cord-derived mesenchymal stem cells alleviate acute lung injury caused by severe burn via secreting TSG-6 and inhibiting inflammatory response | |
| CN102188707B (zh) | Il-17抑制剂在制备治疗流感的药物中的用途 | |
| Wu et al. | Silencing of c-kit with small interference RNA attenuates inflammation in a murine model of allergic asthma | |
| Meng et al. | sTLR4/sMD‑2 complex alleviates LPS‑induced acute lung injury by inhibiting pro‑inflammatory cytokines and chemokine CXCL1 expression |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |