WO2021218594A1 - RÉACTIF ET MÉTHODE DE RÉGULATION DE SOUS UNITÉ β D'INTÉGRINE - Google Patents

RÉACTIF ET MÉTHODE DE RÉGULATION DE SOUS UNITÉ β D'INTÉGRINE Download PDF

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WO2021218594A1
WO2021218594A1 PCT/CN2021/086211 CN2021086211W WO2021218594A1 WO 2021218594 A1 WO2021218594 A1 WO 2021218594A1 CN 2021086211 W CN2021086211 W CN 2021086211W WO 2021218594 A1 WO2021218594 A1 WO 2021218594A1
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integrin
subunit
intestinal
cells
mice
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PCT/CN2021/086211
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Chinese (zh)
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陈剑峰
张海龙
郑雅娟
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中国科学院分子细胞科学卓越创新中心
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    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
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    • 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
    • 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
    • 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
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    • 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
    • C12Q2600/136—Screening for pharmacological compounds
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    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/06—Gastro-intestinal diseases
    • G01N2800/065—Bowel diseases, e.g. Crohn, ulcerative colitis, IBS

Definitions

  • the inventor found that the activation of integrins can be used to regulate immune cell adhesion and homing, the body's intestinal immune homeostasis, and the occurrence and development of intestinal inflammation.
  • the agent that reduces the activity of the integrin ⁇ subunit is selected from:
  • the host cell is a non-human mammalian cell.
  • step (3) Inject the embryonic stem cells obtained in step (2) into mouse blastocysts, and transfer the blastocysts into pseudo-pregnant female mice to obtain chimeric mice,
  • This document also provides a transgenic mouse whose integrin or its ⁇ subunit is inactive or has decreased activity.
  • the intestinal disease benefits from inhibiting the stable adhesion of lymphocytes, reducing the ability of immune cells to migrate across blood vessels, down-regulating the expression of integrin ⁇ E, and inhibiting the return of immune cells to intestinal-related lymphoid tissues.
  • Nest inhibit intestinal inflammation diseases.
  • the disease is intestinal inflammation. More preferably, the intestinal inflammation is chronic intestinal inflammation mediated by adaptive immunity or acute intestinal inflammation mediated by innate immunity.
  • This document also provides a detection kit, which contains reagents for detecting the gene or protein of integrin or its ⁇ subunit.
  • FIG. 5 shows that integrin ⁇ 7 F185A mutation causes a significant decrease in ⁇ E ⁇ 7 + lymphocytes and down-regulates the expression of integrin ⁇ E.
  • A FACS detects the expression of ⁇ E in splenic lymphocytes of WT, +/-, KI and KO mice.
  • B FACS detects the expression of ⁇ E of IEL and LPL in the small intestine and colon of WT, +/-, KI and KO mice.
  • the numbers in the boxes represent the proportion of ⁇ E ⁇ 7 positive cells.
  • TBS 20mM Tris-HCl (pH 7.4), 150mM NaCl, 1mM CaCl 2 , 1mM MgCl 2 ;
  • Cell lysis buffer TBS, 1% Triton X-100, 0.05% Tween 20, Complete Protease Inhibitor Cocktail Tablets, Phosstop Phosphatase Inhibitor Cocktail Tablets;
  • Protein A neutralization buffer 1M Tris-HCl, pH 8.0.
  • Mating offspring of chimeric mice have a certain probability of obtaining KI mice containing F185A point mutations, and then mating with EIIa-Cre tool mice to remove the LoxP sequence.
  • the obtained KI mice need to be continuously mated with C57BL/6J wild-type mice for more than 5 generations to purify the genetic background.
  • Immune cell homing mediated by integrin ⁇ 4 ⁇ 7 plays an important role in the pathogenesis of enteritis. Further, we used the T cell-induced enteritis model to test the ability of KI T cells to induce the onset of enteritis in Rag1 -/- recipient mice. First, CD4 + CD45RB high T cells were obtained by FACS sorting, and they were injected into Rag1 -/- recipient mice through the tail vein. Body weight observation in the subsequent twelve weeks showed that WT CD4 + CD45RB high T cells induced significant weight loss in Rag1 -/- recipient mice, while KI and KO CD4 + CD45RB high T cells could not induce weight loss in Rag1 -/- recipient mice (Figure 7, A).

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  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
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Abstract

La présente invention concerne une méthode de régulation à la baisse de l'expression de l'intégrine αE, de réduction de la capacité de migration des cellules immunitaires à travers les vaisseaux sanguins, d'inhibition de l'adhérence entre une cellule immunitaire et un tissu épithélial, d'inhibition de l'écotropisme de la cellule immunitaire envers un tissu lymphoïde associé à l'intestin, et d'inhibition du séjour de la cellule immunitaire au niveau du tissu lymphoïde associé à l'intestin, et son utilisation, qui comprend la régulation à la baisse de l'expression et de l'activité d'une sous-unité β d'intégrine. La présente invention concerne également l'utilisation d'un réactif dans la préparation d'un médicament destiné à la régulation à la baisse de l'expression de l'intégrine αE, la réduction de la capacité de migration des cellules immunitaires à travers les vaisseaux sanguins, l'inhibition de l'adhérence entre la cellule immunitaire et le tissu épithélial, l'inhibition de l'écotropisme de la cellule immunitaire envers le tissu lymphoïde associé à l'intestin, l'inhibition du séjour de la cellule immunitaire au niveau du tissu lymphoïde associé à l'intestin ou l'inhibition de l'inflammation intestinale.
PCT/CN2021/086211 2020-04-30 2021-04-09 RÉACTIF ET MÉTHODE DE RÉGULATION DE SOUS UNITÉ β D'INTÉGRINE WO2021218594A1 (fr)

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CN202010367306.6A CN113577281A (zh) 2020-04-30 2020-04-30 调控整合素β亚基的试剂和方法

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WO2023169491A1 (fr) * 2022-03-10 2023-09-14 苏州易慕峰生物科技有限公司 Cellule immunitaire à capacité d'adhésion cellulaire régulée à la baisse et utilisation médicale associée

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CN111197057B (zh) * 2018-11-19 2024-04-19 中国科学院分子细胞科学卓越创新中心 调控免疫细胞迁移的组合物和方法

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Title
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YAMEI YU; JIANGHAI ZHU; LI-ZHI MI; THOMAS WALZ; HAO SUN; JIANFENG CHEN; TIMOTHY A SPRINGER: "Structural Specializations of α4β7, an Integrin that Mediates Rolling Adhesion", THE JOURNAL OF CELL BIOLOGY, vol. 196, no. 1, 31 December 2012 (2012-12-31), pages 131 - 146, XP055590148, ISSN: 0021-9525, DOI: 10.1083/jcb.201110023 *
YOUDONG PAN, KUN ZHANG, JUNPENG QI, JIAO YUE, TIMOTHY A. SPRINGER, JIANFENG CHEN: "Cation-π Interaction Regulates Ligand-Binding Affinity and Signaling of Integrin α4β7", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES, vol. 107, no. 50, 14 December 2010 (2010-12-14), pages 21388 - 21393, XP055862618, ISSN: 0027-8424, DOI: 10.1073/pnas.1015487107 *
ZHANG KUN, CHEN JIANFENG: "The Regulation of Integrin Function by Divalent Cations", CELL ADHESION & MIGRATION, vol. 6, no. 1, 1 January 2012 (2012-01-01), pages 20 - 29, XP055862623, ISSN: 1933-6918, DOI: 10.4161/cam.18702 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023169491A1 (fr) * 2022-03-10 2023-09-14 苏州易慕峰生物科技有限公司 Cellule immunitaire à capacité d'adhésion cellulaire régulée à la baisse et utilisation médicale associée

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