Kits and Methods for Assessing Skin Health

Abstract

The invention relates to kits and methods for assessing skin health for a human and the human's susceptibility to skin disorders. The methods involve assessing occurrence in the human's genome of one or more polymorphisms (e.g., single nucleotide polymorphisms) that occur in one or more genes associated disclosed herein and that are associated with a disorder in humans. Preferred assessment and scoring methods are disclosed, as are kits for performing the methods.

1 - 23 . (canceled) 
     
     
         24 . A method of assessing skin health of a human, the method comprising assessing occurrence in the human's genome of disorder-associated polymorphisms in at least two genes selected from the group consisting of
 a) the gene which encodes mitochondrial manganese superoxide dismutase (MnSOD);   b) the gene which encodes cytoplasmic copper/zinc superoxide dismutase (CZSOD);   c) the gene which encodes catalase;   d) the gene which encodes human glutathione peroxidase (hGPX1);   e) the gene which encodes glutathione S transferase P1 (GSTP1);   f) the gene which encodes NAD(P)H:quinone oxidoreductase;   g) the gene which encodes epoxide hydrolase;   h) the gene which encodes tumor necrosis factor alpha (TNF-alpha);   i) the gene which encodes NADH/NADPH oxidase p22 subunit (the phox gene);   j) the gene which encodes nitric oxide synthase;   k) the gene which encodes cytochrome P450;   l) the gene which encodes matrix metalloproteinase 1 (MMP-1); and   m) the gene which encodes profilagrin,   
       whereby occurrence of any of the disorder-associated polymorphisms is an indication that the human has poorer skin health than a human whose genome does not comprise any of the disorder-associated polymorphisms, and whereby occurrence of a plurality of the disorder-associated polymorphisms is an indication that the human has even poorer skin health than a human whose genome does not comprise the disorder-associated polymorphisms, wherein the method further comprises calculating a skin health score by summing, for each of the selected genes in which a disorder-associated polymorphism occurs in the human's genome, the product of a constant and a correlation factor, wherein the correlation factor represents the fraction of humans heterozygous or homozygous for the disorder-associated polymorphism who exhibit the corresponding disorder, whereby the skin health score represents the relative susceptibility of the human to a skin disorder. 
     
     
         25 . The method of  claim 24 , wherein the same constant is used for each selected gene. 
     
     
         26 . A method of assessing skin health of a human, the method comprising assessing occurrence in the human's genome of disorder-associated polymorphisms in at least two genes selected from the group consisting of
 a) the gene which encodes mitochondrial manganese superoxide dismutase (MnSOD);   b) the gene which encodes cytoplasmic copper/zinc superoxide dismutase (CZSOD);   c) the gene which encodes catalase;   d) the gene which encodes human glutathione peroxidase (hGPX1);   e) the gene which encodes glutathione S transferase P1 (GSTP1);   f) the gene which encodes NAD(P)H:quinone oxidoreductase;   g) the gene which encodes epoxide hydrolase;   h) the gene which encodes tumor necrosis factor alpha (TNF-alpha),   i) the gene which encodes NADH/NADPH oxidase p22 subunit (the phox gene);   j) the gene which encodes nitric oxide synthase;   k) the gene which encodes cytochrome P450;   l) the gene which encodes matrix metalloproteinase 1 (MMP-1); and   m) the gene which encodes profilagrin,   
       whereby occurrence of any of the disorder-associated polymorphisms is an indication that the human has poorer skin health than a human whose genome does not comprise any of the disorder-associated polymorphisms, and whereby occurrence of a plurality of the disorder-associated polymorphisms is an indication that the human has even poorer skin health than a human whose genome does not comprise the disorder-associated polymorphisms, wherein each of the disorder-associated polymorphisms for which occurrence is assessed is a single nucleotide polymorphism (SNP). 
     
     
         27 . The method of  claim 26 , wherein occurrence of a SNP is assessed by annealing a nucleic acid derived from the human's genome with a primer that is complementary to the region adjacent the SNP on its 3′ side, extending the primer using a polymerase in order to add a nucleotide residue complementary to the SNP to the primer, and detecting the identity of the nucleotide residue complementary to the SNP. 
     
     
         28 . The method of  claim 27 , wherein the nucleotide residue is a non-extendable residue. 
     
     
         29 . A method of assessing the likelihood that a human will develop a skin disorder, the method comprising assessing occurrence in the human's genome of disorder-associated polymorphisms in at least two genes selected from the group consisting of
 a) the gene which encodes mitochondrial MnSOD;   b) the gene which encodes cytoplasmic CZSOD;   c) the gene which encodes catalase;   d) the gene which encodes hGPX1;   e) the gene which encodes GSTP1;   f) the gene which encodes NAD(P)H:quinone oxidoreductase;   g) the gene which encodes epoxide hydrolase;   h) the gene which encodes TNF-alpha;   i) the phox gene;   j) the gene which encodes nitric oxide synthase;   k) the gene which encodes cytochrome P450;   l) the gene which encodes MMP-1; and   m) the gene which encodes profilagrin,   
       whereby occurrence of any of the disorder-associated polymorphisms is an indication that the human is more susceptible to the skin disorder than a human whose genome does not comprise the disorder-associated polymorphism, and whereby occurrence of a plurality of the disorder-associated polymorphisms is an indication that the human is even more susceptible to the skin disorder than a human whose genome does not comprise the disorder-associated polymorphisms. 
     
     
         30 . The method of  claim 29 , comprising assessing occurrence of at least two disorder-associated polymorphisms selected from the group consisting of
 A) a polymorphism in the open reading frame encoding mitochondrial MnSOD;   B) a polymorphism in the open reading frame encoding cytoplasmic CZSOD;   C) a polymorphism in the promoter region of the gene encoding catalase;   D) a polymorphism in the open reading frame encoding hGPX1;   E) a polymorphism in the open reading frame encoding GSTP1;   F) a polymorphism in the open reading frame encoding NAD(P)H:quinone oxidoreductase;   G) a polymorphism in the open reading frame encoding epoxide hydrolase;   H) a polymorphism in the promoter region of the gene encoding TNF-alpha;   I) a polymorphism in the open reading frame of the phox gene;   J) a polymorphism in the open reading frame encoding nitric oxide synthase;   K) a polymorphism in the 5′ flanking region of the gene encoding cytochrome P 450; and      L) a polymorphism in the promoter region of the gene encoding MMP-1.   
     
     
         31 . The method of  claim 29 , comprising assessing occurrence of a first polymorphism selected from the group consisting of
 i) a polymorphism manifested as occurrence of a codon encoding alanine at amino acid residue 9 of mitochondrial MnSOD;   ii) a polymorphism manifested as occurrence of a codon encoding valine at amino acid residue 9 of mitochondrial MnSOD;   iii) a polymorphism manifested as occurrence of a codon encoding isoleucine at amino acid residue 58 of mitochondrial MnSOD;   iv) a polymorphism manifested as occurrence of a codon encoding thymine at amino acid residue 58 of mitochondrial MnSOD;   v) a polymorphism manifested as occurrence of a codon encoding valine at amino acid residue 7 of cytoplasmic CZSOD;   vi) a polymorphism manifested as occurrence of a codon encoding glutamic acid at amino acid residue 7 of cytoplasmic CZSOD;   vii) a polymorphism manifested as occurrence of a codon encoding cysteine at amino acid residue 6 of cytoplasmic CZSOD;   viii) a polymorphism manifested as occurrence of a codon encoding phenylalanine at amino acid residue 6 of cytoplasmic CZSOD;   ix) a polymorphism manifested as occurrence of a cytosine residue at nucleotide residue  262 of the catalase gene;   x) a polymorphism manifested as occurrence of a thymine residue at nucleotide residue  262 of the catalase gene;   xi) a polymorphism manifested as occurrence of a codon encoding proline at amino acid residue 198 of hGPX1;   xii) a polymorphism manifested as occurrence of a codon encoding leucine at amino acid residue 198 of hGPX1;   xiii) a polymorphism manifested as occurrence of a codon encoding valine at amino acid residue 105 of GSTP1;   xiv) a polymorphism manifested as occurrence of a codon encoding isoleucine at amino acid residue 105 of GSTP1;   xv) a polymorphism manifested as occurrence of a cytosine residue at nucleotide residue 242 of the gene encoding NAD(P)H:quinone oxidoreductase;   xvi) a polymorphism manifested as occurrence of a thymine residue at nucleotide residue 242 of the gene encoding NAD(P)H:quinone oxidoreductase;   xvii) a polymorphism manifested as occurrence of a thymine residue at nucleotide residue 113 in exon 3 of the gene which encodes epoxide hydrolase;   xviii) a polymorphism manifested as occurrence of a cytosine residue at nucleotide residue 113 in exon 3 of the gene which encodes epoxide hydrolase;   xix) a polymorphism manifested as occurrence of an adenine residue at nucleotide residue −238 of the gene which encodes TNF-alpha;   xx) a polymorphism manifested as occurrence of an adenine residue at nucleotide residue −308 of the gene which encodes TNF-alpha;   xxi) a polymorphism manifested as occurrence of a cytosine residue at nucleotide residue 242 of the phox gene;   xxii) a polymorphism manifested as occurrence of a thymine residue at nucleotide residue 242 of the phox gene;   xxiii) a polymorphism manifested as occurrence of a 27 base nucleotide residue between nucleotide residues 5130 and 5511 of the gene encoding nitric oxide synthase;   xxiv) a polymorphism manifested as non-occurrence of a 27 nucleotide residue repeat between nucleotide residues 5130 and 5511 of the gene encoding nitric oxide synthase;   xxv) a polymorphism manifested as occurrence of an adenine residue at nucleotide residue −290 of the gene encoding cytochrome P450;   xxvi) a polymorphism manifested as occurrence of a guanine residue at nucleotide residue −290 of the gene encoding cytochrome P450;   xxvii) a polymorphism manifested as occurrence of a single guanine residue at nucleotide residue −1607 of the human gene encoding MMP-1; and   xxviii) a polymorphism manifested as occurrence of a two consecutive guanine residues at a site including nucleotide residue −1607 of the human gene encoding MMP-1.   
     
     
         32 . A method of selecting a dose of a skin protective composition for administration to a human, the method comprising assessing occurrence in the human's genome of disorder-associated polymorphisms in at least two genes selected from the group consisting of
 a) the gene which encodes mitochondrial MnSOD;   b) the gene which encodes cytoplasmic CZSOD;   c) the gene which encodes catalase;   d) the gene which encodes hGPX1;   e) the gene which encodes GSTP1;   f) the gene which encodes NAJNP)H:quinone oxidoreductase;   g) the gene which encodes epoxide hydrolase;   h) the gene which encodes tumor necrosis factor alpha (TNF-alpha);   i) the phox gene;   j) the gene which encodes nitric oxide synthase;   k) the gene which encodes cytochrome P450;   l) the gene which encodes MMP-1; and   m) the gene which encodes profilagrin,   
       whereby occurrence of any of the disorder-associated polymorphisms is an indication that a greater dose of the composition should be administered to the human; and selecting a dose of the composition based on occurrence of the disorder-associated polymorphisms. 
     
     
         33 . A kit for assessing relative susceptibility of a human to a skin disorder, the kit comprising reagents for assessing occurrence in the human's genome of disorder-associated polymorphisms in at least two genes selected from the group consisting of
 a) the gene which encodes mitochondria' MnSOD;   b) the gene which encodes cytoplasmic CZSOD;   c) the gene which encodes catalase;   d) the gene which encodes hGPX1;   e) the gene which encodes GSTP1;   f) the gene which encodes NAD(P)H:quinone oxidoreductase;   g) the gene which encodes epoxide hydrolase;   h) the gene which encodes tumor necrosis factor alpha (TNF-alpha);   i) the phox gene;   j) the gene which encodes nitric oxide synthase;   k) the gene which encodes cytochrome P450;   l) the gene which encodes MMP-1; and   m) the gene which encodes profilagrin.   
     
     
         34 . The kit of  claim 33 , wherein the reagents comprise first oligonucleotides that anneal with higher stringency with the disorder-associated polymorphisms than with corresponding non-disorder-associated polymorphisms. 
     
     
         35 . The kit of  claim 34 , wherein each of the first oligonucleotides is attached to a support. 
     
     
         36 . The kit of  claim 35 , wherein each of the first oligonucleotides is attached to the same support. 
     
     
         37 . The kit of  claim 35 , wherein each of the first oligonucleotides is attached to a different support. 
     
     
         38 . The kit of  claim 34 , wherein the first oligonucleotides are molecular beacon oligonucleotides. 
     
     
         39 . The kit of  claim 34 , wherein the kit further comprises second oligonucleotides that anneal with higher stringency with the non-disorder-associated polymorphisms than with corresponding disorder-associated polymorphisms. 
     
     
         40 . The kit of  claim 39 , wherein the first and second oligonucleotides are spectrally distinct molecular beacon oligonucleotide pairs. 
     
     
         41 . The kit of  claim 33 , wherein the reagents comprise primers that are complementary to the region adjacent a characteristic residue of the disorder-associated-polymorphism for amplifying at least the characteristic residue. 
     
     
         42 . The kit of  claim 41 , further comprising a polymerase capable of extending the primers by adding a nucleotide residue complementary to the characteristic residue. 
     
     
         43 . The kit of  claim 42 , further comprising a non-extendable nucleotide residue. 
     
     
         44 . The kit of  claim 33 , further comprising an instructional material which includes a numerical value representing the product of a constant and a correlation factor, wherein the correlation factor represents the fraction of humans heterozygous or homozygous for the disorder-associated polymorphism who exhibit the corresponding disorder. 
     
     
         45 . The kit of  claim 44 , wherein the same constant is used for each selected gene. 
     
     
         46 . The kit of  claim 33 , wherein the kit comprises reagents for assessing occurrence in the human's genome of at least two polymorphisms selected from the group consisting of
 I) a polymorphism manifested as a change from an alanine residue to a valine residue at amino acid residue 9 of mitochondrial MnSOD;   II) a polymorphism manifested as a change from an isoleucine residue to a thymine residue at amino acid residue 58 of mitochondrial MnSOD;   III) a polymorphism manifested as a change from a valine residue to a glutamic acid residue at amino acid residue 7 of cytoplasmic CZSOD;   IV) a polymorphism manifested as a change from a cysteine residue to a phenylalanine residue at amino acid residue 6 of cytoplasmic CZSOD;   V) a polymorphism manifested as a change from a cytosine residue to a thymine residue at nucleotide residue −262 of the catalase gene;   VI) a polymorphism in the hGPX1 gene manifested as a change from a proline residue to a leucine residue at amino acid residue 198 of glutathione peroxidase;   VII) a polymorphism in the GSTP1 gene manifested as a change from a valine residue to an isoleucine residue at amino acid residue 105 of glutathione S-transferase P1;   VIII) a polymorphism manifested as a change from a cytosine residue to a thymine residue at nucleotide residue 242 of the gene encoding NAD(P)H:quinone oxidoreductase;   IX) a polymorphism manifested as a change from a thymine residue to a cytosine residue at nucleotide residue 113 in exon 3 of the gene which encodes epoxide hydrolase;   X) a polymorphism manifested as a change to an adenine residue at nucleotide residue  238 of the gene which encodes TNF-alpha;   XI) a polymorphism manifested as a change to an adenine residue at nucleotide residue −308 of the gene which encodes TNF-alpha;   XII) a polymorphism manifested as a change from a cytosine residue to a thymine residue at nucleotide residue 242 of the phox gene;   XIII) a polymorphism manifested as a 27 nucleotide residue repeat between nucleotide residues 5130 and 5511 of the gene encoding nitric oxide synthase;   XIV) a polymorphism manifested as a change from an adenine residue to a guanine residue at nucleotide residue −290 of the gene encoding cytochrome P450; and   XV) a polymorphism manifested as occurrence of two consecutive guanine residues at a site including nucleotide residue −1607 of the human gene encoding MMP-1.   
     
     
         47 . (canceled) 
     
     
         48 . A method of selecting a dose of a skin protective agent for administration to a human in a nutritional product, the method comprising assessing occurrence in the human's genome of disorder-associated polymorphisms in at least two genes selected from the group consisting of
 a) the gene which encodes mitochondrial MnSOD;   b) the gene which encodes cytoplasmic CZSOD;   c) the gene which encodes catalase;   d) the gene which encodes hGPX 1;   e) the gene which encodes GSTP1;   f) the gene which encodes NAD(P)H:quinone oxidoreductase;   g) the gene which encodes epoxide hydrolase;   h) the gene which encodes TNF-alpha;   i) the phox gene;   j) the gene which encodes nitric oxide synthase;   k) the gene which encodes cytochrome P450;   l) the gene which encodes MMP-1; and   m) the gene which encodes profilagrin,   
       whereby occurrence of any of the disorder-associated polymorphisms is an indication that a greater dose of the agent should be administered to the human in the nutritional product; and selecting a dose of the agent for the nutritional product based on occurrence of the disorder-associated polymorphisms.