CN-115698269-B - Adipocytes highly expressing FFAR4 and uses thereof

Abstract

The present invention provides a novel method for the treatment and/or prophylaxis of various diseases, such as age-related impaired glucose tolerance, impaired cognitive ability. The present invention relates to adipocytes that highly express FFAR4 and are useful for the treatment and/or prevention of various diseases, particularly, impaired glucose tolerance associated with aging and impaired cognitive ability associated with aging, a composition for transplantation containing the same, and a method for the treatment and/or prevention of various diseases using the composition for transplantation. Furthermore, the present invention relates to transgenic mice that highly express FFAR 4.

Inventors

• TAKEI YOSHINORI
• Ping Zeming

Assignees

• 株式会社NUMT
• 学校法人东邦大学

Dates

Publication Date

20260505

Application Date

20210526

Priority Date

20200529

Claims (5)

1. 1. Use of adipocytes for the manufacture of a medicament for the treatment and/or prevention of impaired glucose tolerance associated with aging and/or impaired cognitive ability associated with aging, characterized in that, The adipocytes are introduced with a gene encoding FFAR4 and modified to express FFAR4, The adipocytes are obtained by introducing FFAR4 gene into adipose tissue-derived adipose stem cells or adipose precursor cells, forcibly expressing FFAR4, and then inducing differentiation, The adipocytes are produced by a method comprising the steps of: a, preparing a chimeric gene, wherein the chimeric gene is a gene in which a human FFAR4 cDNA is arranged downstream of an appropriate promoter sequence, and NCBI of the human FFAR4 cDNA has accession number NM_181745; b a step of inserting the chimeric gene into a virus or the like and introducing the chimeric gene into a fat stem cell or a fat precursor cell, and C differentiating the fat stem cells or fat precursor cells into which the chimeric gene is introduced into fat cells.
2. 2. The use according to claim 1, wherein, The adipocytes are cells isolated from adipose tissue of a transgenic mouse into which a gene encoding FFAR4 has been introduced.
3. 3. The use according to claim 1, wherein, The promoter sequence is an aP2 gene promoter sequence.
4. 4. The use of a composition for transplantation in the manufacture of a medicament for the treatment and/or prevention of impaired glucose tolerance associated with aging, characterized in that, The composition for transplantation contains adipocytes, into which a gene encoding FFAR4 has been introduced and modified to express FFAR4, The adipocytes are obtained by introducing FFAR4 gene into adipose tissue-derived adipose stem cells or adipose precursor cells, forcibly expressing FFAR4, and then inducing differentiation, The adipocytes are produced by a method comprising the steps of: a, preparing a chimeric gene, wherein the chimeric gene is a gene in which a human FFAR4 cDNA is arranged downstream of an appropriate promoter sequence, and NCBI of the human FFAR4 cDNA has accession number NM_181745; b a step of inserting the chimeric gene into a virus or the like and introducing the chimeric gene into a fat stem cell or a fat precursor cell, and C differentiating the fat stem cells or fat precursor cells into which the chimeric gene is introduced into fat cells.
5. 5. Use of a composition for transplantation in the manufacture of a medicament for the treatment and/or prevention of cognitive impairment associated with aging, characterized in that, The composition for transplantation contains adipocytes, into which a gene encoding FFAR4 has been introduced and modified to express FFAR4, The adipocytes are obtained by introducing FFAR4 gene into adipose tissue-derived adipose stem cells or adipose precursor cells, forcibly expressing FFAR4, and then inducing differentiation, The adipocytes are produced by a method comprising the steps of: a, preparing a chimeric gene, wherein the chimeric gene is a gene in which a human FFAR4 cDNA is arranged downstream of an appropriate promoter sequence, and NCBI of the human FFAR4 cDNA has accession number NM_181745; b a step of inserting the chimeric gene into a virus or the like and introducing the chimeric gene into a fat stem cell or a fat precursor cell, and C differentiating the fat stem cells or fat precursor cells into which the chimeric gene is introduced into fat cells.

Description

Adipocytes highly expressing FFAR4 and uses thereof Technical Field The present invention relates to adipocytes that highly express FFAR4, which are useful in the treatment and/or prevention of various diseases, and to a composition for transplantation containing the same, and to a method for treating and/or preventing various diseases in animals and humans using the composition for transplantation. Examples of the target diseases in the present invention include impaired glucose tolerance associated with age and impaired cognitive function. Background It is known that with age, glucose tolerance is impaired, and the elderly easily develop a state where the blood glucose level after a meal for 2 hours exceeds 140mg/dL (postprandial hyperglycemia), and that the risk of developing diabetes increases. It is considered that prevention of diabetes in the elderly is effective in controlling the rise of postprandial blood glucose level, in other words, in improving impaired glucose tolerance associated with aging, and there is a concern that existing therapeutic agents for diabetes have side effects such as hypoglycemia, and no safe therapeutic method is established at present. The G protein-coupled receptor (GPCR) is a receptor having a characteristic molecular structure penetrating the cell membrane 7 times, and a target molecule of a therapeutic agent for most diseases such as hypertension, arrhythmia, angina pectoris, asthma, peptic ulcer, etc., belongs to the gene family. Thus, GPCRs are attracting attention as drug discovery target molecules. GPCRs are activated from outside cells by specific ligands, transmitting their information to intracellular G proteins to exert physiological activity. The above characteristic molecular structure of GPCRs is believed to undergo a large structural change by binding to a specific ligand, leading to the activation of G proteins. FFAR4, which is one of GPCRs (referred to as gpr120 before the name is specified, and has the same amino acid sequence as registered in GenBank as np_ 859529), was found to be a free fatty acid receptor that activates intracellular information transfer with a ligand that is a free fatty acid (particularly, omega-3 fatty acid called DHA, EPA, which is an essential fatty acid) that is an important nutrient as an energy source, and as its physiological function, a mechanism that promotes intestinal secretion of glucagon-like peptide GLP-1 under stimulation by the free fatty acid was clarified (Hirasawa, a.et al. Na. Med.11,90-94,2005 (non-patent literature 1)). In addition, FFAR4 has been confirmed to interact with long chain fatty acids by an interaction analysis method using a combination of a fluorescent ligand and a receptor of a flow cytometer (Sun, Q et al mol. Pharmacol.78,804-810,2010 (non-patent document 2)). Furthermore, in FFAR4 knockout mice, the phenotype of fat cell hypertrophy, fat tissue weight increase, body weight increase, fatty liver, and abnormal glucose tolerance is exhibited, and this phenomenon is thought to be caused by inhibition of fat tissue differentiation and reduction of fatty acid synthesis (Ichimura, a.et al. Nature 483,350-354,2012 (non-patent document 3)). As described above, FFAR4 has been shown to be closely related to obesity in eating, and for obesity in eating, it is expected to develop FFAR 4-targeted application to preventive/therapeutic drugs (Hara,T.et al.Rev Physiol Biochem Pharmacol.164,77-116,2013.;Milligan,G.et al.Trends Pharmacol Sci.38,809-821,2017.( non-patent documents 4 and 5)). The development of FFAR 4-targeted prophylactic and therapeutic agents has been advanced so far, and there is a trend toward finding compounds having FFAR4 agonist activity, and studies are being made to find compounds having FFAR4 agonist activity useful for the treatment and/or prevention of diabetes, obesity, and hyperlipidemia (japanese patent application laid-open No. 2012-520240 (patent document 1), japanese patent application laid-open No. 2008-001690 (patent document 2), and international publication No. 2005/083070 (patent document 3)). In addition, although cells into which a gene encoding FFAR4 has been introduced have been prepared, no attempt has been made to use cells into which a gene encoding FFAR4 has been introduced for the purpose of screening ligands and analyzing the activity (japanese patent application laid-open publication 2016-214135 (patent document 4), japanese patent application laid-open publication 2012-520240 (patent document 1), and japanese patent application laid-open publication 2008-001690 (patent document 2)). In addition, even for such analysis purposes, FFAR4 analysis in adipocytes is performed using cells obtained by inducing differentiation of mouse-derived cultured cells 3T3-L1 into adipocytes, and as a result, this method uses FFAR4 expression inhibition or the like by induced differentiation, siRNA or the like in addition to an agonist to which FFAR4 is added, and there has been n