Eukaryotic genes involved in adult lifespan regulation

Abstract

The present invention relates to regulation of adult lifespan in eukaryotes. More particularly, the present invention is directed to methods of assaying for genes, gene products, and genes in pathways controlled by such genes and gene products, using RNAi and microarray analysis, that regulate lifespan (e.g., extend or truncate adult lifespan) in eukaryotes such as invertebrates (e.g., C. elegans ), plants, and mammals, e.g., humans. For example, the present invention is directed to genes encoding components of the mitochondrial respiratory chain and genes encoding glycolysis enzymes, which are involved in lifespan regulation, and genes and gene products in pathways controlled by such genes. Other genes and gene products identified as regulating aging and aging pathways include a gene encoding a GTPase; a transcriptional activator; novel genes: llw-1, llw-2, llw-3, and llw-4; genes encoding cytochrome P450 proteins (involved in steroid biosynthesis); a melatonin synthesis gene; genes encoding insulin and insulin-like peptides; genes encoding heat shock factors; genes encoding catalases; stress-response genes; and metabolic genes. The invention further relates to methods for identifying and using agents, including small molecule chemical compositions, antibodies, antisense nucleic acids, and ribozymes, that regulate, e.g., enhance, adult lifespan via modulation of aging associated proteins; as well as to the use of expression profiles, markers, and compositions in diagnosis and therapy related to lifespan extension, life expectancy, and aging. The present invention also relates to gene therapy involving lifespan associated genes.

What is claimed is:  
     
         1 . A method for identifying a compound that modulates aging, the method comprising the steps of: 
 (i) contacting the compound with a polypeptide, wherein the polypeptide is encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid encoding a polypeptide comprising an amino acid sequence selected from the group consisting of: cytochrome C 1  (SEQ ID NO: 1), NADH oxidoreductase (SEQ ID NO: 2), ATP synthase (SEQ ID NO: 3), cytochrome C oxidase (SEQ ID NO: 4), phosphoglucose isomerase (SEQ ID NO: 5), GTPase (SEQ ID NO: 6), LLW-1 (SEQ ID NO: 7), LLW-2 (SEQ ID NO: 8), LLW-3 (SEQ ID NO: 9), LLW-4 (SEQ ID NO: 10) and HSF-1 (SEQ ID NO: 11), and human homologues thereof; and    (ii) determining the functional effect of the compound upon the polypeptide.    
     
     
         2 . The method of  claim 1 , wherein the polypeptide is encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid encoding a polypeptide comprising an amino acid sequence selected from the group consisting of: cytochrome C 1  (SEQ ID NO: 1), NADH oxidoreductase (SEQ ID NO: 2), ATP synthase (SEQ ID NO: 3), cytochrome C oxidase (SEQ ID NO: 4), phosphoglucose isomerase (SEQ ID NO: 5), GTPase (SEQ ID NO: 6), LLW-1 (SEQ ID NO: 7), LLW-2 (SEQ ID NO: 8), LLW-3 (SEQ ID NO: 9), and LLW-4, (SEQ ID NO: 10) or human homologues thereof.  
     
     
         3 . The method of  claim 1 , wherein the polypeptide is encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid encoding a polypeptide comprising an amino acid sequence of HSF-1 (SEQ ID NO: 11), or human homologues thereof.  
     
     
         4 . The method of  claim 1 , wherein the functional effect is determined in vitro.  
     
     
         5 . The method of  claim 4 , wherein the functional effect is determined by measuring enzymatic activity.  
     
     
         6 . The method of  claim 4 , wherein the functional effect is determined by measuring ligand, substrate, or cofactor binding to the polypeptide.  
     
     
         7 . The method of  claim 4 , wherein the functional effect is determined by measuring binding to a promoter sequence by the polypeptide.  
     
     
         8 . The method of  claim 1 , wherein the polypeptide is expressed in a mitochondrial extract or an isolated mitochondrial membrane.  
     
     
         9 . The method of  claim 1 , wherein the polypeptide is expressed in a eukaryotic host or host cell and the polypeptide is contacted with the compound in a living cell.  
     
     
         10 . The method of  claim 9 , wherein the host cell is derived from  C. elegans , mouse, rat, or human.  
     
     
         11 . The method of  claim 9 , wherein the host is  C. elegans , mouse, rat, or human.  
     
     
         12 . The method of  claim 9 , wherein the functional effect is a determined by measuring ligand, substrate, or cofactor binding to the polypeptide.  
     
     
         13 . The method of  claim 9 , wherein the functional effect is determined by measuring transcriptional activation.  
     
     
         14 . The method of  claim 9 , wherein the functional effect is determined by evaluating age-associated parameters.  
     
     
         15 . The method of  claim 9 , wherein the functional effect is determined by evaluating expression of an age-associated gene.  
     
     
         16 . The method of  claim 14 , wherein the age-associated parameter is lifespan.  
     
     
         17 . The method of  claim 1 , wherein the modulation is inhibition of aging.  
     
     
         18 . The method of  claim 17 , wherein inhibition of aging occurs by activation of an HSF polypeptide encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid encoding a polypeptide comprising an amino acid sequence of HSF-1 (SEQ ID NO: 11), or human homologues thereof.  
     
     
         19 . The method of  claim 17 , wherein inhibition of aging occurs by inhibition of a polypeptide encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid encoding a polypeptide comprising an amino acid sequence selected from the group consisting of: cytochrome C 1  (SEQ ID NO: 1), NADH oxidoreductase (SEQ ID NO: 2), ATP synthase (SEQ ID NO: 3), cytochrome C oxidase (SEQ ID NO: 4), phosphoglucose isomerase (SEQ ID NO: 5), GTPase (SEQ ID NO: 6), LLW-1 (SEQ ID NO: 7), LLW-2 (SEQ ID NO: 8), LLW-3 (SEQ ID NO: 9), and LLW-4 (SEQ ID NO: 10) or human homologues thereof.  
     
     
         20 . The method of  claim 1  wherein the polypeptide is recombinant.  
     
     
         21 . The method of  claim 1 , wherein the compound is an antibody, an antisense molecule, or a small molecule.  
     
     
         22 . A method for identifying a compound that modulates aging, the method comprising the steps of: 
 (i) contacting the compound with a polypeptide, wherein the polypeptide is encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid encoding a polypeptide comprising an amino acid sequence selected from the group consisting of: cytochrome C 1  (SEQ ID NO: 1), NADH oxidoreductase (SEQ ID NO: 2), ATP synthase (SEQ ID NO: 3), cytochrome C oxidase (SEQ ID NO: 4), phosphoglucose isomerase (SEQ ID NO: 5), GTPase (SEQ ID NO: 6), LLW-1 (SEQ ID NO: 7), LLW-2 (SEQ ID NO: 8), LLW-3 (SEQ ID NO: 9), LLW-4 (SEQ ID NO: 10) and HSF-1 (SEQ ID NO: 11), and human homologues thereof,    (ii) determining the functional effect of the compound upon the polypeptide; and    (iii) contacting a host or host cell expressing the protein and evaluating an age-associated parameter in a host or host cell, thereby identifying a compound that modulates aging.    
     
     
         23 . The method of  claim 22 , wherein the polypeptide is recombinant.  
     
     
         24 . The method of  claim 22 , wherein the functional effect is a physical effect.  
     
     
         25 . The method of  claim 22 , wherein the functional effect is a chemical effect.  
     
     
         26 . The method of  claim 22 , wherein the functional effect is a phenotypic effect.  
     
     
         27 . The method of  claim 22 , wherein the functional effect is determined in vitro.  
     
     
         28 . The method of  claim 22 , wherein the functional effect is determined in a eukaryotic host organism or host cell.  
     
     
         29 . The method of  claim 22 , wherein the age-associated parameter is lifespan.  
     
     
         30 . A method for identifying a compound that modulates aging, the method comprising the steps of: 
 (i) contacting the compound with a polypeptide that is a component of the mitochondrial respiratory chain; and    (ii) determining the functional effect of the compound upon the polypeptide.    
     
     
         31 . The method of  claim 30 , wherein the polypeptide is expressed in a mitochondrial extract or an isolated mitochondrial membrane.  
     
     
         32 . A method for identifying a compound that modulates aging, the method comprising the steps of: 
 (i) contacting the compound with a polypeptide, wherein the polypeptide is encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid listed in Table 5 or Table 6, or a nucleic acid encoding a polypeptide listed in Table 5 or Table 6, or human homologues thereof; and    (ii) determining the functional effect of the compound upon the polypeptide.    
     
     
         33 . A method for identifying a compound that modulates aging, the method comprising the steps of: 
 (i) contacting the compound with a polypeptide, wherein the polypeptide is encoded by a nucleic acid that hybridizes under stringent conditions to a nucleic acid listed in Table 5 or Table 6, or a nucleic acid encoding a polypeptide listed in Table 5 or Table 6, or human homologues thereof    (ii) determining the functional effect of the compound upon the polypeptide; and    (iii) contacting a host or host cell expressing the protein and evaluating an age-associated parameter in a host or host cell, thereby identifying a compound that modulates aging.    
     
     
         34 . A compound that modulates an aging process, wherein the compound is identified by the method of  claim 1 .  
     
     
         35 . The compound of  claim 34 , wherein the compound is an antibody, an antisense molecule, or a small molecule.  
     
     
         36 . The compound of  claim 34 , wherein the compound is antimycin or an analog thereof.  
     
     
         37 . A method of increasing lifespan or treating premature aging in a subject, the method comprising the step of administering to the subject an effective amount of a compound identified using the method of  claim 1 .  
     
     
         38 . The method of  claim 37 , wherein the compound is antimycin.  
     
     
         39 . A method of increasing lifespan or treating premature aging in a subject, the method comprising the step of administering to the subject an effective amount of a compound that modulates the expression of a polypeptide comprising an amino acid sequence selected from the group consisting of: cytochrome C 1  (SEQ ID NO: 1), NADH oxidoreductase (SEQ ID NO: 2), ATP synthase (SEQ ID NO: 3), cytochrome C oxidase (SEQ ID NO: 4), phosphoglucose isomerase (SEQ ID NO: 5), GTPase (SEQ ID NO: 6), LLW-1 (SEQ ID NO: 7), LLW-2 (SEQ ID NO: 8), LLW-3 (SEQ ID NO: 9), LLW-4 (SEQ ID NO: 10) and HSF-1 (SEQ ID NO: 11), and human homologues thereof.  
     
     
         40 . The method of  claim 37  or  39 , wherein the aging process is abnormal.  
     
     
         41 . The method of  claim 40 , wherein the abnormal aging process is selected from Werner syndrome, Hutchinson-Guilford disease, Bloom's syndrome, Cockayne's syndrome, ataxia telangiectasia, and Down's syndrome.  
     
     
         42 . The method of  claim 37  or  39 , wherein the aging process is normal.  
     
     
         43 . The method of  claim 37  or  39 , further comprising the step of evaluating an age-associated parameter of the subject.  
     
     
         44 . A method of identifying a gene or gene product that modulates aging, the method comprising the steps of: 
 (i) providing a library of nucleic acids, each nucleic acid of the library comprising a segment of a gene of an organism; and    (ii) for each member of the library, 
 (a) generating double-stranded RNA (dsRNA) from the respective member of the library,  
 (b) providing the dsRNA to a cell or one or more cells of the organism to provide a ds-RNA treated cell or ds-RNA treated organism, and  
 (b) monitoring an age-associated parameter of the ds-RNA treated cell or the ds-RNA treated organism.  
   
     
     
         45 . A method of identifying a gene or gene product that modulates aging, the method comprising the steps of: 
 (i) evaluating a relationship between presence or abundance for each species of a plurality of RNA or protein species with respect to age of a cell or organism;    (ii) producing or delivering a double-stranded RNA to a cell or one or more cells of an organism, the RNA corresponding to a species whose presence or abundance is correlated with age; and    (iii) monitoring an age-associated parameter of the cell or the one or more cells or the organism.    
     
     
         46 . A method of identifying a compound that modulates aging, the method comprising the steps of: 
 (i) contacting a test compound to a living or biochemical system that comprising a  C. elegans  target protein selected from the group consisting of: a respiratory chain component, a heat shock promoter activator, LLW-1, LLW-2, LLW-3, LLW-4, a small GTPase, a protein in Table 5, and a protein in Table 6; and    (ii) evaluating a property associated with the target protein; and    (iii) evaluating an aging-associated parameter of a  C. elegans  organism contacted with the test compound.    
     
     
         47 . A method of identifying a gene or gene product that modulates aging, the method comprising the steps of: 
 (i) providing a nematode in which activity of a target protein is reduced in the organism by RNA interference;    (i) expressing a gene encoding a candidate mammalian protein that is heterologous to the organism; and    (iii) evaluating an age associated parameter of the organism.    
     
     
         48 . A  C. elegans  nematode that (1) expresses a heterologous gene in at least some cells, the heterologous gene encoding a respiratory chain component or glycolysis component that is non-identical to the corresponding endogenous respiratory chain component or glycolysis component, or a functional domain thereof; and (2) is deficient in at least some cells for an endogenous activity provided by the corresponding endogenous respiratory chain component or glycolysis component.  
     
     
         49 . A method comprising: assessing an age-associated parameter of the nematode of  claim 48 .  
     
     
         50 . A  C. elegans  nematode that (1) has a deficiency in at least some cells for an endogenous activity, the deficiency generated by dsRNA in the cells, and (2) has an average lifespan of at least 40% greater than an otherwise identical nematode without the deficiency.  
     
     
         51 . A method of identifying a gene or gene product that modulates aging, the method comprising the steps of: 
 (i) providing the nematode of  claim 50;     (ii) introducing a heterologous gene that encodes a heterologous polypeptide into the nematode;    (iii) expressing the heterologous gene in the nematode or a progeny of the nematode under conditions wherein the heterologous polypeptide is produced; and    (iv) monitoring an age-associated parameter of the nematode or the progeny of the nematode.    
     
     
         52 . The method of  claim 51 , further comprising contacting a test compound to the nematode or the progeny prior to or during the monitoring.