CN113332417A - Application of TREM-2 in preparation of tumor treatment medicine and/or diagnostic reagent - Google Patents
Application of TREM-2 in preparation of tumor treatment medicine and/or diagnostic reagent Download PDFInfo
- Publication number
- CN113332417A CN113332417A CN202110460789.9A CN202110460789A CN113332417A CN 113332417 A CN113332417 A CN 113332417A CN 202110460789 A CN202110460789 A CN 202110460789A CN 113332417 A CN113332417 A CN 113332417A
- Authority
- CN
- China
- Prior art keywords
- trem
- tumor
- cells
- gene
- expression
- 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.)
- Withdrawn
Links
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 83
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 10
- 101710174937 Triggering receptor expressed on myeloid cells 2 Proteins 0.000 title claims abstract 16
- 102100029678 Triggering receptor expressed on myeloid cells 2 Human genes 0.000 title claims abstract 14
- 238000002360 preparation method Methods 0.000 title claims description 4
- 239000003814 drug Substances 0.000 title abstract description 4
- 230000014509 gene expression Effects 0.000 claims abstract description 29
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims abstract description 22
- 201000005202 lung cancer Diseases 0.000 claims abstract description 22
- 208000020816 lung neoplasm Diseases 0.000 claims abstract description 22
- 210000001744 T-lymphocyte Anatomy 0.000 claims abstract description 16
- 208000010507 Adenocarcinoma of Lung Diseases 0.000 claims abstract description 12
- 201000005249 lung adenocarcinoma Diseases 0.000 claims abstract description 12
- 201000007270 liver cancer Diseases 0.000 claims abstract description 11
- 208000014018 liver neoplasm Diseases 0.000 claims abstract description 11
- 201000001441 melanoma Diseases 0.000 claims abstract description 10
- 206010009944 Colon cancer Diseases 0.000 claims abstract description 9
- 208000029742 colonic neoplasm Diseases 0.000 claims abstract description 9
- 238000003745 diagnosis Methods 0.000 claims abstract description 8
- 208000003950 B-cell lymphoma Diseases 0.000 claims abstract description 5
- 206010042971 T-cell lymphoma Diseases 0.000 claims abstract description 5
- 208000027585 T-cell non-Hodgkin lymphoma Diseases 0.000 claims abstract description 5
- 210000004027 cell Anatomy 0.000 claims description 70
- 108090000623 proteins and genes Proteins 0.000 claims description 21
- 101000946889 Homo sapiens Monocyte differentiation antigen CD14 Proteins 0.000 claims description 19
- 102100035877 Monocyte differentiation antigen CD14 Human genes 0.000 claims description 18
- 230000000694 effects Effects 0.000 claims description 14
- 210000001616 monocyte Anatomy 0.000 claims description 10
- 230000002401 inhibitory effect Effects 0.000 claims description 7
- 238000013518 transcription Methods 0.000 claims description 7
- 230000035897 transcription Effects 0.000 claims description 7
- 206010025323 Lymphomas Diseases 0.000 claims description 5
- 206010061309 Neoplasm progression Diseases 0.000 claims description 5
- 230000005751 tumor progression Effects 0.000 claims description 5
- 230000004913 activation Effects 0.000 claims description 3
- 241000713666 Lentivirus Species 0.000 claims description 2
- 239000012190 activator Substances 0.000 claims description 2
- 230000000259 anti-tumor effect Effects 0.000 claims description 2
- 230000000295 complement effect Effects 0.000 claims description 2
- 230000002950 deficient Effects 0.000 claims description 2
- 239000002773 nucleotide Substances 0.000 claims description 2
- 125000003729 nucleotide group Chemical group 0.000 claims description 2
- 239000012268 protein inhibitor Substances 0.000 claims description 2
- 229940121649 protein inhibitor Drugs 0.000 claims description 2
- 230000006798 recombination Effects 0.000 claims description 2
- 238000005215 recombination Methods 0.000 claims description 2
- 238000012546 transfer Methods 0.000 claims description 2
- 241000701161 unidentified adenovirus Species 0.000 claims description 2
- 208000019691 hematopoietic and lymphoid cell neoplasm Diseases 0.000 claims 2
- 210000002540 macrophage Anatomy 0.000 abstract description 32
- 210000004881 tumor cell Anatomy 0.000 abstract description 21
- 206010057249 Phagocytosis Diseases 0.000 abstract description 13
- 230000008782 phagocytosis Effects 0.000 abstract description 13
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 abstract description 8
- 210000000066 myeloid cell Anatomy 0.000 abstract description 5
- 238000009169 immunotherapy Methods 0.000 abstract description 3
- 210000004369 blood Anatomy 0.000 abstract description 2
- 239000008280 blood Substances 0.000 abstract description 2
- 230000001413 cellular effect Effects 0.000 abstract description 2
- 231100000433 cytotoxic Toxicity 0.000 abstract description 2
- 230000001472 cytotoxic effect Effects 0.000 abstract description 2
- 229940079593 drug Drugs 0.000 abstract description 2
- 239000003596 drug target Substances 0.000 abstract description 2
- 230000002147 killing effect Effects 0.000 abstract description 2
- 239000003147 molecular marker Substances 0.000 abstract description 2
- 241000699670 Mus sp. Species 0.000 description 30
- 241000699666 Mus <mouse, genus> Species 0.000 description 28
- 102100034922 T-cell surface glycoprotein CD8 alpha chain Human genes 0.000 description 24
- 238000002474 experimental method Methods 0.000 description 20
- 238000001514 detection method Methods 0.000 description 16
- 238000010186 staining Methods 0.000 description 12
- 230000001965 increasing effect Effects 0.000 description 10
- 241001465754 Metazoa Species 0.000 description 8
- 210000005259 peripheral blood Anatomy 0.000 description 8
- 239000011886 peripheral blood Substances 0.000 description 8
- 238000007920 subcutaneous administration Methods 0.000 description 8
- 210000001519 tissue Anatomy 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 7
- 238000011160 research Methods 0.000 description 7
- 238000011813 knockout mouse model Methods 0.000 description 6
- 230000004614 tumor growth Effects 0.000 description 6
- 210000001266 CD8-positive T-lymphocyte Anatomy 0.000 description 5
- 230000006870 function Effects 0.000 description 5
- 238000003197 gene knockdown Methods 0.000 description 5
- 230000012010 growth Effects 0.000 description 5
- 210000004698 lymphocyte Anatomy 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 4
- 206010061218 Inflammation Diseases 0.000 description 4
- 230000003213 activating effect Effects 0.000 description 4
- 238000001727 in vivo Methods 0.000 description 4
- 230000004054 inflammatory process Effects 0.000 description 4
- 210000004072 lung Anatomy 0.000 description 4
- 230000000242 pagocytic effect Effects 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 102000005962 receptors Human genes 0.000 description 4
- 108020003175 receptors Proteins 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 108060003951 Immunoglobulin Proteins 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 239000012091 fetal bovine serum Substances 0.000 description 3
- 238000010166 immunofluorescence Methods 0.000 description 3
- 102000018358 immunoglobulin Human genes 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 210000000274 microglia Anatomy 0.000 description 3
- 238000002054 transplantation Methods 0.000 description 3
- 208000024827 Alzheimer disease Diseases 0.000 description 2
- 201000011240 Frontotemporal dementia Diseases 0.000 description 2
- 208000018737 Parkinson disease Diseases 0.000 description 2
- 210000000683 abdominal cavity Anatomy 0.000 description 2
- 239000000556 agonist Substances 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 208000015114 central nervous system disease Diseases 0.000 description 2
- 238000013399 early diagnosis Methods 0.000 description 2
- 239000006274 endogenous ligand Substances 0.000 description 2
- 210000004013 groin Anatomy 0.000 description 2
- 208000024200 hematopoietic and lymphoid system neoplasm Diseases 0.000 description 2
- 230000028993 immune response Effects 0.000 description 2
- 230000002757 inflammatory effect Effects 0.000 description 2
- 230000028709 inflammatory response Effects 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 201000010901 lateral sclerosis Diseases 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 208000005264 motor neuron disease Diseases 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 230000019491 signal transduction Effects 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 1
- 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 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 238000011746 C57BL/6J (JAX™ mouse strain) Methods 0.000 description 1
- 210000001239 CD8-positive, alpha-beta cytotoxic T lymphocyte Anatomy 0.000 description 1
- 102000029816 Collagenase Human genes 0.000 description 1
- 108060005980 Collagenase Proteins 0.000 description 1
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 1
- 108010020382 Hepatocyte Nuclear Factor 1-alpha Proteins 0.000 description 1
- 108010061414 Hepatocyte Nuclear Factor 1-beta Proteins 0.000 description 1
- 102100022057 Hepatocyte nuclear factor 1-alpha Human genes 0.000 description 1
- 102100022123 Hepatocyte nuclear factor 1-beta Human genes 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101100025714 Homo sapiens NLRP12 gene Proteins 0.000 description 1
- 101000795117 Homo sapiens Triggering receptor expressed on myeloid cells 2 Proteins 0.000 description 1
- 102000037982 Immune checkpoint proteins Human genes 0.000 description 1
- 108091008036 Immune checkpoint proteins Proteins 0.000 description 1
- 108090001005 Interleukin-6 Proteins 0.000 description 1
- 102000004889 Interleukin-6 Human genes 0.000 description 1
- 108010090054 Membrane Glycoproteins Proteins 0.000 description 1
- 102000012750 Membrane Glycoproteins Human genes 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 101000795119 Mus musculus Triggering receptor expressed on myeloid cells 3 Proteins 0.000 description 1
- 101150104437 NLRP12 gene Proteins 0.000 description 1
- 102000011779 Nitric Oxide Synthase Type II Human genes 0.000 description 1
- 108010076864 Nitric Oxide Synthase Type II Proteins 0.000 description 1
- 108090000412 Protein-Tyrosine Kinases Proteins 0.000 description 1
- 102000004022 Protein-Tyrosine Kinases Human genes 0.000 description 1
- 208000007660 Residual Neoplasm Diseases 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 208000005718 Stomach Neoplasms Diseases 0.000 description 1
- 108010066451 Triggering Receptor Expressed on Myeloid Cells-1 Proteins 0.000 description 1
- 102000018368 Triggering Receptor Expressed on Myeloid Cells-1 Human genes 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000001363 autoimmune Effects 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 210000004979 bone marrow derived macrophage Anatomy 0.000 description 1
- 230000011748 cell maturation Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 229960002424 collagenase Drugs 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 210000005220 cytoplasmic tail Anatomy 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 210000004443 dendritic cell Anatomy 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 201000002491 encephalomyelitis Diseases 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000005714 functional activity Effects 0.000 description 1
- 206010017758 gastric cancer Diseases 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 102000046699 human CD14 Human genes 0.000 description 1
- 210000003917 human chromosome Anatomy 0.000 description 1
- 230000004727 humoral immunity Effects 0.000 description 1
- 229940126546 immune checkpoint molecule Drugs 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 229940100601 interleukin-6 Drugs 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000031146 intracellular signal transduction Effects 0.000 description 1
- 210000003125 jurkat cell Anatomy 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 229920006008 lipopolysaccharide Polymers 0.000 description 1
- 210000005265 lung cell Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000005087 mononuclear cell Anatomy 0.000 description 1
- 210000000822 natural killer cell Anatomy 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 210000002997 osteoclast Anatomy 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 230000005885 phagocytic elimination Effects 0.000 description 1
- DCWXELXMIBXGTH-UHFFFAOYSA-N phosphotyrosine Chemical compound OC(=O)C(N)CC1=CC=C(OP(O)(O)=O)C=C1 DCWXELXMIBXGTH-UHFFFAOYSA-N 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 201000011549 stomach cancer Diseases 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000008467 tissue growth Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 102000027257 transmembrane receptors Human genes 0.000 description 1
- 108091008578 transmembrane receptors Proteins 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
Images
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
- A61K38/1774—Immunoglobulin superfamily (e.g. CD2, CD4, CD8, ICAM molecules, B7 molecules, Fc-receptors, MHC-molecules)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- A61K48/005—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57419—Specifically defined cancers of colon
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57423—Specifically defined cancers of lung
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57438—Specifically defined cancers of liver, pancreas or kidney
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
- G01N33/57492—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving compounds localized on the membrane of tumor or cancer cells
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Urology & Nephrology (AREA)
- Molecular Biology (AREA)
- Hematology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Food Science & Technology (AREA)
- Microbiology (AREA)
- Oncology (AREA)
- Hospice & Palliative Care (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Gastroenterology & Hepatology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to an application of TREM-2 in preparing tumor treatment medicines and/or diagnostic reagents, belongs to the technical field of cellular immunotherapy, and provides an application prospect of a myeloid cell trigger receptor (TREM-2) in tumor treatment, wherein according to the two aspects, on one hand, TREM-2 is highly expressed and enhances the phagocytosis and killing effect of mononuclear macrophages on tumor cells, and on the other hand, TREM-2 is highly expressed and promotes the cytotoxic effect of CD8+ T lymphocytes on tumor cells. The TREM-2 expression level increase in peripheral blood mononuclear cells or CD8+ T lymphocytes of various tumor patients can be used as a molecular marker for tumor diagnosis. The invention can provide good candidate drug targets and diagnosis indexes for clinical treatment of blood system tumors such as lung cancer, B cell lymphoma, T cell lymphoma and the like, and solid tumors such as colon cancer, liver cancer, melanoma, lung adenocarcinoma and the like, and has very good application prospects.
Description
Technical Field
The invention relates to the technical field of cellular immunotherapy, and in particular relates to application of TREM-2 in preparation of tumor treatment medicines and/or diagnostic reagents.
Background
The incidence and mortality of tumors are rapidly increasing worldwide, and cancer is one of the leading causes of death and is also a significant barrier to improving human life expectancy in countries around the world. Myeloid cell-Triggered Receptors (TREMs) are a family of immunoglobulin receptors expressed on the cell surface of myeloid cells, and the genes encoding them are located on human chromosome 6p21 and murine chromosome 17C 3. TREM receptors include TREM-1 and TREM-2 in humans, TREM-3 in mice, and at least two other TREM-like related molecules (TREM-like transcription factor-1 and TREM-like transcription factor-2). TREM-2 is an important member having a central role in the TREM family, is a membrane protein encoded by the TREM-2 gene, belongs to the transmembrane receptor of the TREM protein family, which is a glycosylated single-channel type I membrane glycoprotein comprising extracellular immunoglobulin domain, transmembrane domain and cytoplasmic tail 3 portion, which can undergo intracellular signal transduction by DNAX activating protein 12 and phosphorylated tyrosine kinase, the expression of which is seen on macrophages, microglia and osteoclasts. TREM-2 stimulates the production of inflammatory factors in the chronic inflammatory progression and immune response, which are associated with a variety of functions such as cell maturation, survival, proliferation, activation, phagocytosis, etc.
Endogenous ligands and agonists of TREM-2 are not discovered at present, and the mechanism of signal transduction is not completely defined. Recent gene screening results prove that the TREM-2 gene heterozygosis mutation is a risk factor of Alzheimer disease, Parkinson disease, frontotemporal dementia and lateral sclerosis. Researches show that TREM-2 can inhibit macrophage from secreting tumor necrosis factor alpha, interleukin-6 and other proinflammatory factors, so as to inhibit macrophage activity and weaken immune response of the macrophage to lipopolysaccharide. Overexpression of TREM-2 in microglia reduces the expression of TNF- α and inducible nitric oxide synthase in the meta-co-cultured cells. Inhibition of TREM-2 expression exacerbates autoimmune encephalomyelitis. Together, these studies suggest that TREM-2 exists as an inflammation inhibitor in the inflammatory response. Modulation and expression of TREM-2 has been of interest to a large number of researchers, particularly immunologists and pharmacologists, and has become a potential target for the treatment of inflammation-related central nervous system disorders.
At present, the expression change and functional activity of TREM-2 in malignant tumors are rarely studied, and particularly, the function of TREM-2 on tumor-specific killer T lymphocytes is not reported to be studied at present. Current immunotherapy for tumors still has great application limitations and ineffectiveness. In order to improve the early diagnosis and non-operative treatment effects of tumors and the survival rate of tumor patients, a new immune checkpoint molecule is urgently needed to be found to improve the early diagnosis rate and the immune treatment effect of tumors.
Disclosure of Invention
In order to overcome the problems in the prior art, the invention provides a new application of a myeloid cell Triggering receptor-2 (TREM-2).
The purpose of the invention is realized by the following technical scheme:
application of TREM-2 in preparing an anti-tumor functional product, wherein the functional product has an effect of inhibiting tumor progression, and the tumor is a blood tumor and a solid tumor.
Preferably, the tumor is selected from lung cancer, T-cell lymphoma, B-cell lymphoma, colon cancer, liver cancer, melanoma, lung adenocarcinoma.
Myeloid cell-triggered receptor-2 (TREM-2) is a member of the immunoglobulin superfamily that is mainly expressed on the cell surfaces of monocytes, macrophages, microglia, dendritic cells, natural killer cells, etc., and consists of extracellular immunoglobulin-like regions, transmembrane regions, and intracellular regions. At present, the endogenous ligand and agonist of TREM-2 are not discovered, and the signal transduction mechanism is not completely clear. Recent gene screening results prove that the TREM-2 gene heterozygosis mutation is a risk factor of Alzheimer disease, Parkinson disease, frontotemporal dementia and lateral sclerosis. Together, these studies suggest that TREM-2 exists as an inflammation inhibitor in the inflammatory response. Modulation and expression of TREM-2 has been of interest to a large number of researchers, particularly immunologists and pharmacologists, and has become a potential target for the treatment of inflammation-related central nervous system disorders. According to the invention, research on tumor cells shows that the phagocytosis and elimination of the tumor cells by TREM-2 knocked-out macrophages are obviously weakened, the tumor progression can be promoted, and TREM-2 is prompted to have the effect of inhibiting the tumor progression; in addition to the research on the phagocytosis of tumor cells, the invention also researches the TREM-2 in a tumor microenvironment to regulate the function state of T lymphocytes, and the immunofluorescence examination finds that TREM-2 molecules are highly expressed in CD8+ T cells in the tumor microenvironment, and in an animal experiment for tumor modeling, the growth size of the tumor of a TREM-2 full-knock mouse is obviously larger than that of a wild mouse, and the TREM-2 has the effect of inhibiting the tumor progression, so that the TREM-2 is expected to be applied to the comprehensive treatment of the tumor.
Preferably, the functional product has a function of up-regulating the expression, transcription, or an expression product thereof of the TREM-2 gene.
More preferably, the expression product of the TREM-2 gene refers to various forms of molecules of the NLRP12 gene at various stages, such as but not limited to molecules produced by the TREM-2 gene during amplification, replication, transcription, splicing, processing, translation, modification, such as cDNA, mRNA, precursor protein, mature protein, and fragments thereof.
As a preferred embodiment, the functional product includes: one or more of a TREM-2 protein inhibitor, a TREM-2 gene deficient or silenced immune-related cell, a differentiated cell thereof, or a gene recombination construct.
More preferably, the functional product comprises:
(i) an activated antibody, nucleotide, lentivirus or adenovirus which takes a TREM-2 transcript as a target sequence and can activate the expression of a TREM-2 gene expression product or gene transcription;
(ii) a construct containing a TREM-2 complementary sequence and capable of forming an activator molecule which promotes expression of a TREM-2 gene expression product or gene transcription upon transfer into the body;
(iii) immune-related cells, differentiated cells thereof, or constructs following activation of a TREM-2 gene sequence.
On the one hand, the research of the invention finds that: (1) the TREM-2 expression level is increased in peripheral blood mononuclear cells of a lung cancer patient, (2) the TREM-2 expression level is increased in spleen mononuclear cells of a lung cancer mouse, (3) the TREM-2 expression level is increased in macrophages of the lung cancer mouse, (4) the subcutaneous tumor volume of the TREM-2 knockout mouse on the macrophages is remarkably larger than that of an unbundled mouse, and (5) the phagocytic elimination capacity of the TREM-2 knockout macrophages on tumor cells is remarkably weakened.
Therefore, in practical application, TREM-2 can be over-expressed in macrophages to enhance the phagocytic and scavenging capacity of cells on tumor cells, so that the purpose of treating tumors is achieved. As a preferred embodiment, the application may specifically be: the TREM-2 activating antibody is used for activating TREM-2 on macrophages in vivo, or preparing TREM-2 high-expression bionic macrophages or CAR-T, and the TREM-2 activating antibody has the effect of targeted treatment of tumors.
In another aspect of the invention, the research also finds that: the expression of TREM-2 on CD8+ T cells in peripheral blood mononuclear cells of a tumor patient is obviously higher than that of a normal human (P < 0.0001); in human tumor tissue sections, immunofluorescence examination finds that TREM-2 molecules are highly expressed in CD8+ T cells in a tumor microenvironment; in tumor-modeled animal experiments, we found that TREM-2 full knock-out mice had significantly larger tumor growth size than wild-type mice (P < 0.05). Therefore, the TREM-2 can be over-expressed on the CD8+ T cells and then is returned to the body to enhance the humoral immunity, so that the aim of treating the tumor is fulfilled.
From the two aspects of research, the invention also provides application of TREM-2 as a target spot in preparing a reagent for diagnosing tumors, wherein the tumors are blood tumors and solid tumors.
Preferably, the tumor is selected from T-cell lymphoma, B-cell lymphoma, colon cancer, liver cancer, melanoma, lung adenocarcinoma.
Preferably, TERM-2 is expressed on the surface of CD8+ T lymphocytes, or on the surface of CD14 monocytes; the tumor diagnostic reagent is used for detecting the expression level of TERM-2 on the surface of CD8+ T lymphocytes or on the surface of CD14 monocytes.
Compared with the prior art, the invention has the following beneficial effects:
the invention provides an application prospect of a myeloid cell trigger receptor (TREM-2) in tumor treatment, which comprises two aspects, on one hand, TREM-2 is highly expressed and enhances the phagocytosis and killing effect of mononuclear macrophages on tumor cells, and on the other hand, TREM-2 is highly expressed and promotes the cytotoxic effect of CD8+ T lymphocytes on tumor cells. The TREM-2 expression level increase in peripheral blood mononuclear cells or CD8+ T lymphocytes of various tumor patients can be used as a molecular marker for tumor diagnosis. The invention can provide good candidate drug targets and diagnosis indexes for clinical treatment of blood system tumors such as lung cancer, B cell lymphoma, T cell lymphoma and the like, and solid tumors such as colon cancer, liver cancer, melanoma, lung adenocarcinoma and the like, and has very good application prospects.
Drawings
FIG. 1 shows in vitro phagocytosis experiments that TREM-2 knockdown significantly reduces the phagocytic capacity of macrophages to tumor cells, and TREM-2f/f-Lyz2-Cre mice are TREM-2 knockdown mice on macrophages; in the figure, represents P <0.05, represents P <0.01, represents P < 0.001;
FIG. 2 shows in vivo phagocytosis experiments that TREM-2 knockdown significantly reduces the phagocytic capacity of macrophages to tumor cells, and TREM-2f/f-Lyz2-Cre mice are TREM-2 knockdown mice on macrophages;
FIG. 3 shows subcutaneous transplantation tumor experiment showing that the subcutaneous transplantation tumor volume of TREM-2 knockout mouse on macrophage is significantly increased, and TREM-2f/f-Lyz2-Cre mouse is TREM-2 knockout mouse on macrophage;
FIG. 4 shows that the TREM-2 has a significantly increased positive proportion of CD14 monocytes in peripheral blood of patients with lung cancer;
FIG. 5 is a statistical scatter plot of flow analyzer detection of TREM-2 in CD8+ T lymphocytes; the percentage of TREM-2 and CD8 double positive cells in CD8 cells was detected;
FIG. 6 shows that TREM-2 has a significantly increased positive proportion of F4/80 macrophages in lung cancer mice;
FIG. 7 shows an immunofluorescence plot of TREM-2 expression on CD8+ T lymphocytes in tumor tissue sections from a colon cancer patient, with TREM-2 found to be highly expressed on CD8+ T lymphocytes in the tumor microenvironment; (in the figure, blue represents DAPI, red represents TREM-2 molecules, green represents CD8 molecules, the upper row represents observation under a low power lens, and the lower row represents observation under a high power lens);
FIG. 8 is a tumor growth curve for a set of mouse melanoma subcutaneous modeling experiments, with white circles in the lower panel representing tumor growth volume for wild type mice and black squares representing tumor growth volume for TREM-2 full knock mice; (in the figure, represents P <0.05, represents P <0.01, represents P < 0.001);
FIG. 9 is a set of mouse liver cancer subcutaneous modeling animal experiments, wherein A is a comparison of tumor tissue growth curves, white circles represent tumor growth volumes of wild-type mice, and black boxes represent tumor growth volumes of TREM-2 full-knock mice; b is the appearance of tumor tissue; graph C represents the weight of subcutaneous tumor tissue after stripping, (graph x represents P <0.05, P < 0.001).
Detailed Description
The following further describes the embodiments of the present invention. It should be noted that the description of the embodiments is provided to help understanding of the present invention, but the present invention is not limited thereto. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
The test methods used in the following experimental examples are all conventional methods unless otherwise specified; the materials, reagents and the like used are, unless otherwise specified, commercially available reagents and materials.
Example 1 in vitro phagocytosis assay investigating the Effect of TREM-2 on phagocytosis of tumor cells by macrophages
1. Collecting a detection sample:
co-incubation cultured macrophages and tumor cells were collected.
2. The detection method specifically comprises the following steps:
(1) bone marrow-derived macrophage BMDM (from C57BL/6J WT or TREM-2f/f-Lyz2-Cre mice) cells at 1X10524-well plates were inoculated per well density and cultured overnight.
(2) The next day the medium was discarded, washed twice with 1 × PBS, and starved for 2 hours with serum-free basal DMEM medium. Taking mouse lung adenocarcinoma cell LLC as an example, 2x10 was added after 2 hours5Individual CFSE-labeled tumor cells were co-cultured in a 37 degree incubator for 2 hours.
(3) Then all cells were collected by digestion and flow stained for F4/80 (phagocytosis rate: percentage of CFSE + F4/80+ double positive cells); BMDM cells were washed completely and observed for adhesion by inverted fluorescence microscopy (phagocytosis rate: ratio of the number of green fluorescent tumor cells contained in 100 macrophages).
3. The experimental results are as follows:
the results are shown in FIG. 1, and the phagocytic capacity of TREM-2 knocked-out macrophages on a series of tumor cells (A20: mouse B lymphoma cells, Raji: human B lymphoma cells, Jurkat: human T lymphoma cells, LLC: mouse lung adenocarcinoma cells, A549: human lung adenocarcinoma cells) is obviously reduced, which indicates that TREM-2 has the effect of inhibiting tumors under in vitro conditions.
Example 2 in vivo phagocytosis assay investigating the Effect of TREM-2 on phagocytosis of tumor cells by macrophages
1. Collecting a detection sample:
the remaining tumor cells in the abdominal cavity were collected.
2. The detection method specifically comprises the following steps:
(1) CFSE-labeled mouse lung adenocarcinoma cell LLC at 1X107One mouse/mouse was intraperitoneally injected into C57BL/6J WT or TREM-2f/f-Lyz2-Cre mice.
(2) Mice were sacrificed by 24 hour posterior cervical dislocation, the remaining tumor cells in the abdominal cavity were collected, centrifuged at 1500 rpm for 5 minutes, and then washed with 1mL of PBS.
(3) CollectingAll cells were subjected to flow cytometry, the Remaining CFSE-labeled tumor cells were quantified, and the ratio of Remaining cells, Remaining LLC cells (%) -, was calculated (Remaining LLC cells/1X 10)7)×100％。
3. The experimental results are as follows:
the result is shown in figure 2, the proportion of residual tumor cells on macrophages in TREM-2 knockout mice is obviously higher than that of WT mice, and the TREM-2 also has the function of inhibiting tumors in vivo.
Example 3 comparison of size of experimental tumors of subcutaneous transplantation of TREM-2 knockout mice
1. Collecting a detection sample:
a subcutaneous transplanted tumor model was constructed, tumor size was recorded every two days, and growth volume was recorded.
2. The detection method specifically comprises the following steps:
(1) mouse lung adenocarcinoma cell LLC at 1x106One/side was inoculated subcutaneously in the groin of C57BL/6J WT or TREM-2f/f-Lyz2-Cre mice, bilaterally.
(2) On day 7, the long and short diameters of the tumor were measured with a vernier caliper every two days after the tumor started to grow, and the state of the mouse was observed.
(3) Recording was stopped by day 19 before tumor diameter exceeded animal ethics and mice were sacrificed by cervical dislocation.
3. The experimental results are as follows:
the result is shown in figure 3, the tumor volume of the TREM-2f/f-Lyz2-Cre mouse is obviously larger than that of the WT mouse, and the TREM-2 on the macrophage has an inhibiting effect on the growth of the tumor.
Example 4 comparison of the Positive proportion of TREM-2 monocytes in peripheral blood CD14 from patients with Lung cancer
1. Collecting a detection sample:
subject peripheral blood was collected from 36 healthy subjects and 28 lung cancer patients.
2. The detection method specifically comprises the following steps:
(1) separating PBMC from peripheral blood with human lymphocyte separation solution, collecting 1 × 105～1×107Adding PBS into the cellsCentrifuge at 1500 rpm for 5 minutes in 1 mL.
(2) Specific antibody staining reaction: a100. mu.L reaction system contained: PBMC is 1X105～1×107The cells, PBS 100. mu.L, TREM-2 antibody or isotype control antibody IgG 3. mu.L, CD14 antibody 2. mu.L, were incubated on ice (0-4 ℃) for 30 minutes in the absence of light. PBS 1mL was added and centrifuged at 1500 rpm for 5 minutes, which was repeated 3 times.
(3) Add PBS 500uL heavy suspension cells, flow analyzer detection.
(4) And (4) judging a result: and (4) taking the isotype control antibody as negative, judging non-specific staining, if no obvious staining effect exists, indicating that the non-specific staining can be ignored, and then counting positive cells. The percentage of TREM-2 and CD14 double positive cells to CD14 single positive cells was calculated.
3. The experimental results are as follows:
the results are shown in fig. 4, and among the CD14 monocytes of 28 lung cancer patients, the percentage of TREM-2 and CD14 double positive cells in CD14 single positive cells was more than 95% on average; of the 36 healthy human CD14 monocytes, all healthy human TREM-2 and CD14 double positive cells accounted for less than 80% of the CD14 single positive cells on average. The results show that TREM-2 on the surface of the CD14 monocyte of the lung cancer patient is remarkably increased, and the kit can be used for diagnosis and treatment of the lung cancer.
Example 5 Positive proportion test of TREM-2 on peripheral blood CD8 lymphocytes of tumor patients
1. Collecting a detection sample:
peripheral blood of the tested subject is collected, 134 healthy people are included, and 51 tumor patients including lung cancer, liver cancer, colon cancer, leukemia, breast cancer and gastric cancer are included.
2. The detection method specifically comprises the following steps:
separating PBMC from peripheral blood with lymphocyte separation solution, collecting 1 × 105～1x107Adding 1ml of PBS into each cell, and centrifuging for 5 minutes at 1500 rpm;
(1) specific antibody staining reaction: the 100ul reaction system contained: PBMC is 1X105～1x107Individual cells, PBS100ul, TREM-2 antibody or isotype control antibody IgG 3ul, CD14 antibody 2ul, CD8 antibody 2ul, placing the reaction system on ice (0-4 ℃) and incubating for 30 minutes in the dark. Adding 1ml of PBS, centrifuging at 1500 rpm for 5 minutes, and repeating for 3 times;
(2) adding 500ul PBS to resuspend the cells, and detecting by a flow analyzer;
3. and (4) judging a result: and (4) taking the isotype control antibody as negative, judging non-specific staining, if no obvious staining effect exists, indicating that the non-specific staining can be ignored, and then counting positive cells. The percentage of TREM-2 and CD8 double positive cells to CD8 single positive cells was calculated. The percentage of TREM-2 and CD14 double positive cells to CD14 single positive cells was calculated.
4. The experimental results are as follows:
the results in fig. 5 show that of the CD8+ T lymphocytes from 50 tumor patients, 48 of TREM-2 and CD8 double positive cells accounted for more than 30% of the single positive cells of CD8 with an accuracy of 96%; among 125 healthy human CD8 lymphocytes, the percentage of TREM-2 and CD8 double-positive cells in all healthy human CD8 single-positive cells is less than 30%, and the accuracy rate reaches 100%. The above results indicate that TREM-2 on the surface of CD8 lymphocytes can effectively distinguish healthy people from tumor patients.
In conclusion, in peripheral blood of a tumor patient, TREM-2 on the surface of CD8+ T lymphocytes is remarkably increased compared with that of healthy people, and the diagnosis of the tumor can be performed by analyzing TREM-2 on the surface of CD8+ T lymphocytes, so that the accuracy can reach 96%, and the method has the advantages of high sensitivity and good specificity.
Example 6 comparison of the Positive proportion of F4/80 macrophages in Lung cancer mice by TREM-2
1. Collecting a detection sample:
experimental mouse lung tissues were collected, including 5 mice each for control mice and lung cancer modeling mice.
2. The detection method specifically comprises the following steps:
(1) mouse lung adenocarcinoma cell LLC at 1x106One/side was inoculated subcutaneously in the groin of C57BL/6J WT mice, bilaterally. Mice were observed daily until tumor diameter exceeded that of the animalsRearing was stopped before ethical regulation and mice were sacrificed by cervical dislocation.
(2) Collecting lung of mouse, digesting with collagenase, grinding, collecting 1 × 105～1×107The cells were centrifuged at 1500 rpm for 5 minutes with 1mL PBS added.
(3) Specific antibody staining reaction: a100. mu.L reaction system contained: lung cell suspension is 1X105～1×107And (3) incubating the cells, 100 mu L of PBS, 3 mu L of TREM-2 antibody or isotype control antibody IgG, and 2 mu L of F4/80 antibody on ice (0-4 ℃) for 30 minutes in a dark place. PBS 1mL was added and centrifuged at 1500 rpm for 5 minutes, which was repeated 3 times.
(4) Add PBS 500uL heavy suspension cells, flow analyzer detection.
(5) And (4) judging a result: and (4) taking the isotype control antibody as negative, judging non-specific staining, if no obvious staining effect exists, indicating that the non-specific staining can be ignored, and then counting positive cells. The percentage of TREM-2 and F4/80 double positive cells to F4/80 single positive cells was calculated.
3. The experimental results are as follows:
the result is shown in figure 6, the percentage of TREM-2 and F4/80 double positive cells in lung cancer mouse lung macrophages is close to 95% of the percentage of F4/80 single positive cells; in the macrophages in the lungs of the control mice, the percentage of TREM-2 and F4/80 double positive cells to F4/80 single positive cells averaged less than 85%. The results show that TREM-2 on the macrophage surface of the lung cancer mouse is obviously increased, and the lung cancer mouse can be used for diagnosis and treatment of lung cancer.
Example 7 detection of the expression of TREM-2 on CD8+ T cells in human tumor tissue sections
1. Experimental Material
Paraffin embedded human colon cancer tissue samples.
2. Experimental methods
The specific operation is shown in Table 1.
TABLE 1
3. Results of the experiment
The results of the experiment are shown in FIG. 7. The results show that TREM-2 is highly expressed on CD8+ T lymphocytes in the tumor tissue microenvironment.
Example 8 animal experiments on melanoma
1. Experimental Material
(1) Cell lines: mouse melanoma cells (B16 cells) were cultured in DMEM medium (containing 10% fetal bovine serum).
(2) Experimental animals: male C57BL/6J TREM-2 gene knockdown mice (6-8 weeks) and littermate control wild type C57BL/6J mice of the same sex and week age were housed in SPF-grade environment.
2. Experimental methods
(1) Cell culture:
the cells were cultured in DMEM medium (containing 10% fetal bovine serum, pH 7.2) supplemented with 2mmol/L glutamine, and cultured in a cell culture incubator at 37 ℃ under 5% CO 2.
3. Animal experiments:
4. Results of the experiment
The results of the experiment are shown in FIG. 8, which shows: TREM-2 inhibits the growth of melanoma in mice.
Example 9 animal experiments on liver cancer
1. Experimental Material
(1) Cell lines: mouse liver cancer cells (Hepa1-6 cells) were cultured in DMEM medium (containing 10% fetal bovine serum).
(2) Experimental animals: see example 3.
2. Experimental methods
(1) See example 3 for cell culture.
(2) Animal experiments:
physiological saline containing 5X 10^5 Hepa1-6 cells was injected subcutaneously into the right inguinal of the mouse, and the tumor size was measured every two days when the tumor grew and could be measured (the long and short diameters of the tumor were measured, and the tumor volume was 1/2X long diameter X short diameter ^ 2). Mice were sacrificed on day 16.
3. Results of the experiment
The results of the experiment are shown in FIG. 9, which shows: TREM-2 can inhibit the growth of liver cancer in mice.
Claims (9)
- The application of TREM-2 in preparing an anti-tumor functional product is characterized in that the functional product has the effect of inhibiting tumor progression, and the tumor is a hematological tumor and a solid tumor.
- 2. Use according to claim 1, wherein said tumor is selected from lung cancer, T-lymphoma, B-lymphoma, colon cancer, liver cancer, melanoma, lung adenocarcinoma.
- 3. The use according to claim 1, wherein the functional product has a function of up-regulating TREM-2 gene expression, transcription, or an expression product thereof.
- 4. The use according to claim 3, wherein the functional product comprises: one or more of a TREM-2 protein inhibitor, a TREM-2 gene deficient or silenced immune-related cell, a differentiated cell thereof, or a gene recombination construct.
- 5. The use according to claim 4, wherein the functional product comprises:(i) an activated antibody, nucleotide, lentivirus or adenovirus which takes a TREM-2 transcript as a target sequence and can activate the expression of a TREM-2 gene expression product or gene transcription;(ii) a construct containing a TREM-2 complementary sequence and capable of forming an activator molecule which promotes expression of a TREM-2 gene expression product or gene transcription upon transfer into the body;(iii) immune-related cells, differentiated cells thereof, or constructs following activation of a TREM-2 gene sequence.
- Use of TREM-2 as a target in the preparation of a reagent for the diagnosis of a tumor, wherein the tumor is a hematological tumor or a solid tumor.
- 7. The use according to claim 6, wherein said tumor is selected from the group consisting of lung cancer, T-cell lymphoma, B-cell lymphoma, colon cancer, liver cancer, melanoma, lung adenocarcinoma.
- 8. The use of claim 7, wherein TERM-2 is expressed on the surface of CD8+ T lymphocytes or on the surface of CD14+ monocytes.
- 9. The use of claim 7, wherein said reagent is used to detect the expression level of TERM-2 on the surface of CD8+ T lymphocytes or on the surface of CD14 monocytes.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110460789.9A CN113332417A (en) | 2021-04-27 | 2021-04-27 | Application of TREM-2 in preparation of tumor treatment medicine and/or diagnostic reagent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110460789.9A CN113332417A (en) | 2021-04-27 | 2021-04-27 | Application of TREM-2 in preparation of tumor treatment medicine and/or diagnostic reagent |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113332417A true CN113332417A (en) | 2021-09-03 |
Family
ID=77468762
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110460789.9A Withdrawn CN113332417A (en) | 2021-04-27 | 2021-04-27 | Application of TREM-2 in preparation of tumor treatment medicine and/or diagnostic reagent |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113332417A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2715100A1 (en) * | 2001-03-20 | 2002-09-20 | Novo Nordisk A/S | A novel receptor trem(triggering receptor expressed on myeloid cells) and uses thereof |
| US20100299767A1 (en) * | 2008-02-22 | 2010-11-25 | Industry Foundation Of Chonnam National University | Trem-2 gene and protein as inhibitors of expression of ga733-2, and transgenic animals comprising the same and uses thereof |
| CN104093424A (en) * | 2011-09-30 | 2014-10-08 | 中外制药株式会社 | Antigen-binding molecule inducing immune response to target antigen |
| WO2019070755A1 (en) * | 2017-10-02 | 2019-04-11 | The Broad Institute, Inc. | Methods and compositions for detecting and modulating an immunotherapy resistance gene signature in cancer |
-
2021
- 2021-04-27 CN CN202110460789.9A patent/CN113332417A/en not_active Withdrawn
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2715100A1 (en) * | 2001-03-20 | 2002-09-20 | Novo Nordisk A/S | A novel receptor trem(triggering receptor expressed on myeloid cells) and uses thereof |
| US20100299767A1 (en) * | 2008-02-22 | 2010-11-25 | Industry Foundation Of Chonnam National University | Trem-2 gene and protein as inhibitors of expression of ga733-2, and transgenic animals comprising the same and uses thereof |
| CN104093424A (en) * | 2011-09-30 | 2014-10-08 | 中外制药株式会社 | Antigen-binding molecule inducing immune response to target antigen |
| WO2019070755A1 (en) * | 2017-10-02 | 2019-04-11 | The Broad Institute, Inc. | Methods and compositions for detecting and modulating an immunotherapy resistance gene signature in cancer |
Non-Patent Citations (2)
| Title |
|---|
| AITOR ESPARZA-BAQUER等: "TREM-2 defends the liver against hepatocellular carcinoma through multifactorial protective mechanisms", GUT, vol. 70, no. 7, pages 1345 * |
| 姚一楠: "髓系细胞触发受体2 (TREM-2)在肺癌中的负向免疫调控作用及机制研究", 中国博士学位论文全文数据库（电子期刊）医药卫生科技辑 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Lodolce et al. | T cell–independent interleukin 15rα signals are required for bystander proliferation | |
| Konjevic et al. | Investigation of NK cell function and their modulation in different malignancies | |
| KR101518409B1 (en) | A method for expanding monocytes | |
| Frassanito et al. | Deregulated cytokine network and defective Th1 immune response in multiple myeloma | |
| Ding et al. | CD40 controls CXCR5-induced recruitment of myeloid-derived suppressor cells to gastric cancer | |
| EP1875246B1 (en) | Diagnostic method for detecting cancer by measuring amount of a cytokine like il-6 | |
| KR20200065026A (en) | Methods for isolating T cells with antigen specificity for P53 cancer-specific mutations | |
| Weinberg et al. | The generation of T cell memory: a review describing the molecular and cellular events following OX40 (CD134) engagement | |
| TWI572718B (en) | Immunosuppressive cells and making methods and composition thereof | |
| EP2275114B1 (en) | REIC protein for use in activating cancer immunity | |
| EP2139498B1 (en) | Methods for detecting a natural killer (nk) cell-mediated immune response and an increase in nk cell activity | |
| CN109265563B (en) | human chimeric antigen receptor for treating blood tumor and its application | |
| US7659084B2 (en) | Methods for detecting and isolating antigen-specific T lymphocytes with CD40/C154 inhibitors | |
| Cloosen et al. | Mucin-1 is expressed on dendritic cells, both in vitro and in vivo | |
| CN113332417A (en) | Application of TREM-2 in preparation of tumor treatment medicine and/or diagnostic reagent | |
| CN110585427B (en) | Composition for improving immunity of organism and application of composition in resisting adult T cell leukemia or nasopharyngeal carcinoma | |
| Jovanović et al. | Th-17 cells as novel participants in immunity to breast cancer | |
| Iranparast et al. | Essential Transcription Factors and Functional Roles of Follicular Helper T Cells in Human Autoimmune Diseases | |
| CN110709508A (en) | Methods of treating neogenetic diseases | |
| WO2023080210A1 (en) | Identification of treatment target in aggressive nk leukemia | |
| Zhao et al. | Blockade of OX40/OX40L signaling using anti-OX40L delays disease progression in murine lupus | |
| Yang et al. | Research and Analysis of Molecules such as CD28, CD45RA, CD45RO, CD38, HLA-DR, and CD57 on T Cells in Multiple Myeloma. | |
| Karatza | Investigating the role of Atypical Chemokine Receptor 3 in cancer development | |
| CN114657123A (en) | Exosome cell-free vaccine of over-expression RAE-1 derived from leukemia specific dendritic cell and preparation method thereof | |
| Bendriss-Vermare et al. | Interleukin-35 impairs human NK cell effector functions and induces their ILC1-like conversion with tissue residency features |
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 | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20210903 |
|
| WW01 | Invention patent application withdrawn after publication |