DNA Damage; DNA Repair; Melanoma, Experimental; Molecular Biology; Skin Neoplasms; Sunburn; Xeroderma Pigmentosum; Mutagenesis; Photobiology; DNA Adducts
Therapeutic Radiology: Radiobiology
Skin Diseases Research Center, Yale
Cancer begins as an encounter between a carcinogen and a gene. We are pinpointing these early events, which occur decades before the appearance of a tumor. Our past work on sunlight-induced mutations in tumor suppressor genes has led us to three current topics: 1) Exploring how UVB-induced apoptosis and UV-driven cell fate decisions drive a single mutant cell to clonally expand. 2) Determining rates of DNA photoproduct formation and repair across the genome, as dosimeters of a person's past UV exposure. 3) The interaction of UV and melanin in causing melanoma.
Extensive Research Description
The story thus far, from photons up to cells: UV leads to mutations at the site of DNA photoproducts (rather than elevating genomic instability); the important photoproducts are cyclobutane dimers and (6-4) photoproducts, which join adjacent cytosines or thymines; only the cytosine mutates; these unique properties create a characteristic "mutation signature" for UV that can be seen in tumors decades later; sunlight mutates the P53 and PTCH genes in non-melanoma skin cancer; P53 is required for UV-induced apoptosis, which prevents mutations; apoptosis is signaled by DNA photoproducts in actively transcribed genes and by a product of UV-irradiated melanin; another cause of apoptosis is exposure of melanin to sunlight, particularly the melanin found in blonde and red hair; and our sun-exposed skin carries about 60,000 tiny clones of P53-mutant keratinocytes. Expansion of single mutant cells into clones is due to physiology rather than a 2nd mutation: UV-induced apoptosis deletes normal progenitor cells while sparing the mutant ones. UV also tilts the progenitor cell's fate decision toward self-renewal rather than differentiation. Now, the lab is in the midst of some "functional genomics" questions:
- How does a single mutant keratinocyte expand into a clone? We are attempting to track the clonal expansion of mutant stem cells in a living mouse.
- What DNA regions are the most important UV targets? In addition to actively-transcribed genes, we find that telomeres are UV-sensitive and fail to repair this damage. We've developed a method to measure UV photoproducts and repair in specific genomic regions such as telomeres, and across the entire genome.
- Is cancer development in melanoma unique because of its melanin? We are looking at how reactive oxygen and nitrogen species triggered by UV exposure and reacting with melanin affect genetic and epigenetic fidelity, as well as whether protection against these events varies between individuals due to DNA polymorphisms.
Chemiexcitation of melanin derivatives induces DNA photoproducts long after UV exposure.
Premi, S, Wallisch, S, Mano, CM, Weiner, AB, Bacchiocchi, A, Wakamatsu, K, Bechara, EJH, Halaban, R, Douki, T, and Brash, DE. Chemiexcitation of melanin derivatives induces DNA photoproducts long after UV exposure. Science 347:842-7, 2015.
Human telomeres are hypersensitive to UV-induced DNA damage and refractory to repair.
Rochette, PJ and Brash, DE. Human telomeres are hypersensitive to UV-induced DNA damage and refractory to repair. PLoS Genetics 6 (e1000926): 1-13, 2010.
Stochastic fate of p53-mutant epidermal progenitor cells is tilted toward proliferation by UVB during preneoplasia.
Klein, AM, Brash, DE, Jones, PH, and Simons, BD. Stochastic fate of p53-mutant epidermal progenitor cells is tilted toward proliferation by UVB during preneoplasia. Proc. Natl. Acad. Sci. USA. 107:270-275, 2010.
Bcl-2 is the target of a UV-inducible apoptosis switch and a node for UV signaling.
Knezevic, D, et al. Bcl-2 is the target of a UV-inducible apoptosis switch and a node for UV signaling. Proc. Natl. Acad. Sci. USA. 104:11286-91, 2007.
Melanin acts as a potent UVB sensitizer to cause an atypical mode of cell death in murine skin.
Takeuchi, S, Zhang, W, Wakamatsu, K, Ito, S, Hearing, V, Kraemer, KH, and Brash, DE. Melanin acts as a potent UVB sensitizer to cause an atypical mode of cell death in murine skin. Proc. Natl. Acad. Sci. USA, 101:15076-15081, 2004.
Inactivating E2f1 reverts apoptosis resistance and cancer sensitivity in Trp53-deficient mice.
Wikonkal, NM, Remenyik, E, Knezevic, D., Zhang, W, Liu, M, Zhou, H, Berton, TR, Johnson, DG, and Brash, DE. Inactivating E2f1 reverts apoptosis resistance and cancer sensitivity in Trp53-deficient mice. Nature Cell Biol. 5:655-660, 2003.
Full List of PubMed Publications
- Anand S, Rollakanti KR, Brankov N, Brash DE, Hasan T, Maytin EV: Fluorouracil Enhances Photodynamic Therapy of Squamous Cell Carcinoma via a p53-Independent Mechanism that Increases Protoporphyrin IX levels and Tumor Cell Death. Mol Cancer Ther. 2017 Jun; 2017 Mar 23. PMID: 28336806
- Premi S, Brash DE: Chemical excitation of electrons: A dark path to melanoma. DNA Repair (Amst). 2016 Aug; 2016 Jun 1. PMID: 27262612
- Brash DE: UV-induced Melanin Chemiexcitation: A New Mode of Melanoma Pathogenesis. Toxicol Pathol. 2016 Jun; 2016 Mar 7. PMID: 26951162
- Premi S, Brash DE: Unanticipated role of melanin in causing carcinogenic cyclobutane pyrimidine dimmers. Mol Cell Oncol. 2016 Jan; 2015 Jun 23. PMID: 27308551
- Brash DE: Cancer. Preprocancer. Science. 2015 May 22. PMID: 25999495
- Premi S, Wallisch S, Mano CM, Weiner AB, Bacchiocchi A, Wakamatsu K, Bechara EJ, Halaban R, Douki T, Brash DE: Photochemistry. Chemiexcitation of melanin derivatives induces DNA photoproducts long after UV exposure. Science. 2015 Feb 20. PMID: 25700512
- Brash DE: UV signature mutations. Photochem Photobiol. 2015 Jan-Feb; 2014 Nov 28. PMID: 25354245
- Buschmann T, Zhang R, Brash DE, Bystrykh LV: Enhancing the detection of barcoded reads in high throughput DNA sequencing data by controlling the false discovery rate. BMC Bioinformatics. 2014 Aug 7; 2014 Aug 7. PMID: 25099007
- Laurence M, Hatzis C, Brash DE: Common contaminants in next-generation sequencing that hinder discovery of low-abundance microbes. PLoS One. 2014; 2014 May 16. PMID: 24837716
- Zhang R, Premi S, Kilic SS, Bacchiocchi A, Halaban R, Brash DE: Clonal growth of human melanocytes using cell-free extracellular matrix. Pigment Cell Melanoma Res. 2013 Nov; 2013 Sep 19. PMID: 24034857
- Krauthammer M, Kong Y, Ha BH, Evans P, Bacchiocchi A, McCusker JP, Cheng E, Davis MJ, Goh G, Choi M, Ariyan S, Narayan D, Dutton-Regester K, Capatana A, Holman EC, Bosenberg M, Sznol M, Kluger HM, Brash DE, Stern DF, Materin MA, Lo RS, Mane S, Ma S, Kidd KK, Hayward NK, Lifton RP, Schlessinger J, Boggon TJ, Halaban R: Exome sequencing identifies recurrent somatic RAC1 mutations in melanoma. Nat Genet. 2012 Sep; 2012 Jul 29. PMID: 22842228
- Rochette PJ, Brash DE: Human telomeres are hypersensitive to UV-induced DNA Damage and refractory to repair. PLoS Genet. 2010 Apr 29; 2010 Apr 29. PMID: 20442874
- Klein AM, Brash DE, Jones PH, Simons BD: Stochastic fate of p53-mutant epidermal progenitor cells is tilted toward proliferation by UV B during preneoplasia. Proc Natl Acad Sci U S A. 2010 Jan 5; 2009 Dec 15. PMID: 20018764
- Rochette PJ, Lacoste S, Therrien JP, Bastien N, Brash DE, Drouin R: Influence of cytosine methylation on ultraviolet-induced cyclobutane pyrimidine dimer formation in genomic DNA. Mutat Res. 2009 Jun 1; 2009 Feb 28. PMID: 19427505
- Chao DL, Eck JT, Brash DE, Maley CC, Luebeck EG: Preneoplastic lesion growth driven by the death of adjacent normal stem cells. Proc Natl Acad Sci U S A. 2008 Sep 30; 2008 Sep 24. PMID: 18815380
- Rochette PJ, Brash DE: Progressive apoptosis resistance prior to senescence and control by the anti-apoptotic protein BCL-xL. Mech Ageing Dev. 2008 Apr; 2008 Jan 4. PMID: 18262222
- Knezevic D, Zhang W, Rochette PJ, Brash DE: Bcl-2 is the target of a UV-inducible apoptosis switch and a node for UV signaling. Proc Natl Acad Sci U S A. 2007 Jul 3; 2007 Jun 22. PMID: 17586682
- Brash DE: Roles of the transcription factor p53 in keratinocyte carcinomas. Br J Dermatol. 2006 May. PMID: 16712710
- Raj D, Brash DE, Grossman D: Keratinocyte apoptosis in epidermal development and disease. J Invest Dermatol. 2006 Feb. PMID: 16418733
- Asplund A, Gustafsson AC, Wikonkal NM, Sela A, Leffell DJ, Kidd K, Lundeberg J, Brash DE, Pontén F: PTCH codon 1315 polymorphism and risk for nonmelanoma skin cancer. Br J Dermatol. 2005 May. PMID: 15888139
- Brash DE, Zhang W, Grossman D, Takeuchi S: Colonization of adjacent stem cell compartments by mutant keratinocytes. Semin Cancer Biol. 2005 Apr. PMID: 15652454
- Zhang W, Hanks AN, Boucher K, Florell SR, Allen SM, Alexander A, Brash DE, Grossman D: UVB-induced apoptosis drives clonal expansion during skin tumor development. Carcinogenesis. 2005 Jan; 2004 Oct 21. PMID: 15498793
- Takeuchi S, Zhang W, Wakamatsu K, Ito S, Hearing VJ, Kraemer KH, Brash DE: Melanin acts as a potent UVB photosensitizer to cause an atypical mode of cell death in murine skin. Proc Natl Acad Sci U S A. 2004 Oct 19; 2004 Oct 11. PMID: 15477596
- Knezevic D, Brash DE: Role of E2F1 in apoptosis: a case study in feedback loops. Cell Cycle. 2004 Jun; 2004 Jun 6. PMID: 15107604
- Remenyik E, Wikonkál NM, Zhang W, Paliwal V, Brash DE: Antigen-specific immunity does not mediate acute regression of UVB-induced p53-mutant clones. Oncogene. 2003 Sep 25. PMID: 14508517
- Brash DE: Photocopying cancer cells. J Invest Dermatol. 2003 Jul. PMID: 12839596
- Wikonkal NM, Remenyik E, Knezevic D, Zhang W, Liu M, Zhao H, Berton TR, Johnson DG, Brash DE: Inactivating E2f1 reverts apoptosis resistance and cancer sensitivity in Trp53-deficient mice. Nat Cell Biol. 2003 Jul. PMID: 12833065
- Xue D, Shi H, Smith JD, Chen X, Noe DA, Cedervall T, Yang DD, Eynon E, Brash DE, Kashgarian M, Flavell RA, Wolin SL: A lupus-like syndrome develops in mice lacking the Ro 60-kDa protein, a major lupus autoantigen. Proc Natl Acad Sci U S A. 2003 Jun 24; 2003 Jun 3. PMID: 12788971
- Crasto CJ, Morse TM, Migliore M, Nadkarni P, Hines M, Brash DE, Miller PL, Shepherd GM: Creating knowledgebases to text-mine PUBMED articles using clustering techniques. AMIA Annu Symp Proc. 2003. PMID: 14728326
- Brash DE, Havre PA: New careers for antioxidants. Proc Natl Acad Sci U S A. 2002 Oct 29; 2002 Oct 21. PMID: 12391310
- Havre PA, O'Reilly S, McCormick JJ, Brash DE: Transformed and tumor-derived human cells exhibit preferential sensitivity to the thiol antioxidants, N-acetyl cysteine and penicillamine. Cancer Res. 2002 Mar 1. PMID: 11888918
- Zhang W, Remenyik E, Zelterman D, Brash DE, Wikonkal NM: Escaping the stem cell compartment: sustained UVB exposure allows p53-mutant keratinocytes to colonize adjacent epidermal proliferating units without incurring additional mutations. Proc Natl Acad Sci U S A. 2001 Nov 20; 2001 Nov 13. PMID: 11707578
- Brash DE, Wikonkal NM, Remenyik E, van der Horst GT, Friedberg EC, Cheo DL, van Steeg H, Westerman A, van Kranen HJ: The DNA damage signal for Mdm2 regulation, Trp53 induction, and sunburn cell formation in vivo originates from actively transcribed genes. J Invest Dermatol. 2001 Nov. PMID: 11710938
- Grossman D, Kim PJ, Blanc-Brude OP, Brash DE, Tognin S, Marchisio PC, Altieri DC: Transgenic expression of survivin in keratinocytes counteracts UVB-induced apoptosis and cooperates with loss of p53. J Clin Invest. 2001 Oct. PMID: 11581300
- Liu M, Wikonkal NM, Brash DE: Induction of cyclin-dependent kinase inhibitors and G(1) prolongation by the chemopreventive agent N-acetylcysteine. Carcinogenesis. 1999 Sep. PMID: 10469636
- Wikonkal NM, Brash DE: Ultraviolet radiation induced signature mutations in photocarcinogenesis. J Investig Dermatol Symp Proc. 1999 Sep. PMID: 10537000
- Liu M, Wikonkal NM, Brash DE: UV induces p21WAF1/CIP1 protein in keratinocytes without p53. J Invest Dermatol. 1999 Aug. PMID: 10469320
- Liu M, Pelling JC, Ju J, Chu E, Brash DE: Antioxidant action via p53-mediated apoptosis. Cancer Res. 1998 Apr 15. PMID: 9563490
- Brash DE, Pontén J: Skin precancer. Cancer Surv. 1998. PMID: 10489624
- Brash DE: Sunlight and the onset of skin cancer. Trends Genet. 1997 Oct. PMID: 9351343
- Jonason AS, Kunala S, Price GJ, Restifo RJ, Spinelli HM, Persing JA, Leffell DJ, Tarone RE, Brash DE: Frequent clones of p53-mutated keratinocytes in normal human skin. Proc Natl Acad Sci U S A. 1996 Nov 26. PMID: 8943054
- Gailani MR, Ståhle-Bäckdahl M, Leffell DJ, Glynn M, Zaphiropoulos PG, Pressman C, Undén AB, Dean M, Brash DE, Bale AE, Toftgård R: The role of the human homologue of Drosophila patched in sporadic basal cell carcinomas. Nat Genet. 1996 Sep. PMID: 8782823
- Leffell DJ, Brash DE: Sunlight and skin cancer. Sci Am. 1996 Jul. PMID: 8658110
- Brash DE: Cellular proofreading. Nat Med. 1996 May. PMID: 8616708
- Brash DE, Ziegler A, Jonason AS, Simon JA, Kunala S, Leffell DJ: Sunlight and sunburn in human skin cancer: p53, apoptosis, and tumor promotion. J Investig Dermatol Symp Proc. 1996 Apr. PMID: 9627707
- Ziegler A, Jonason A, Simon J, Leffell D, Brash DE: Tumor suppressor gene mutations and photocarcinogenesis. Photochem Photobiol. 1996 Apr. PMID: 8934758
- Gailani MR, Leffell DJ, Ziegler A, Gross EG, Brash DE, Bale AE: Relationship between sunlight exposure and a key genetic alteration in basal cell carcinoma. J Natl Cancer Inst. 1996 Mar 20. PMID: 8609643
- Brash DE, Gilchrest BA: Photobiology. Wrinkles waiting for GoDoT. Nature. 1996 Jan 25. PMID: 8552182
- Kunala S, Brash DE: Intragenic domains of strand-specific repair in Escherichia coli. J Mol Biol. 1995 Feb 17. PMID: 7869378
- Haffty BG, Carter D, Flynn SD, Fischer DB, Brash DE, Simons J, Ziegler AM, Fischer JJ: Local recurrence versus new primary: clinical analysis of 82 breast relapses and potential applications for genetic fingerprinting. Int J Radiat Oncol Biol Phys. 1993 Oct 20. PMID: 8226151
- Kunala S, Brash DE: Excision repair at individual bases of the Escherichia coli lacI gene: relation to mutation hot spots and transcription coupling activity. Proc Natl Acad Sci U S A. 1992 Nov 15. PMID: 1438309
- Reiss M, Brash DE, Muñoz-Antonia T, Simon JA, Ziegler A, Vellucci VF, Zhou ZL: Status of the p53 tumor suppressor gene in human squamous carcinoma cell lines. Oncol Res. 1992. PMID: 1486218
- Brash DE, Rudolph JA, Simon JA, Lin A, McKenna GJ, Baden HP, Halperin AJ, Pontén J: A role for sunlight in skin cancer: UV-induced p53 mutations in squamous cell carcinoma. Proc Natl Acad Sci U S A. 1991 Nov 15. PMID: 1946433
- Brash DE: Infectious paradigm. Nature. 1991 May 2. PMID: 2027389
- McDonald JW, Brash DE, Oseroff AR, Harris CC: Photosensitizing dyes for the selection of nontumorigenic revertants from human lung cancer cell lines. Cancer Res. 1990 Sep 1. PMID: 2386942
- Mitchell DL, Brash DE, Nairn RS: Rapid repair kinetics of pyrimidine(6-4)pyrimidone photoproducts in human cells are due to excision rather than conformational change. Nucleic Acids Res. 1990 Feb 25. PMID: 2315046
- Garvey N, Witkin EM, Brash DE: Ultraviolet photoproducts at the ochre suppressor mutation site in the glnU gene of Escherichia coli: relevance to "mutation frequency decline". Mol Gen Genet. 1989 Nov. PMID: 2695824
- Kaighn ME, Reddel RR, Lechner JF, Peehl DM, Camalier RF, Brash DE, Saffiotti U, Harris CC: Transformation of human neonatal prostate epithelial cells by strontium phosphate transfection with a plasmid containing SV40 early region genes. Cancer Res. 1989 Jun 1. PMID: 2541897
- Brash DE: UV mutagenic photoproducts in Escherichia coli and human cells: a molecular genetics perspective on human skin cancer. Photochem Photobiol. 1988 Jul. PMID: 3064116
- Reddel RR, Ke Y, Gerwin BI, McMenamin MG, Lechner JF, Su RT, Brash DE, Park JB, Rhim JS, Harris CC: Transformation of human bronchial epithelial cells by infection with SV40 or adenovirus-12 SV40 hybrid virus, or transfection via strontium phosphate coprecipitation with a plasmid containing SV40 early region genes. Cancer Res. 1988 Apr 1. PMID: 2450641
- Harris CC, Brash DE, Lechner JF, Mark G: Aberrations of growth and differentiation pathways during neoplastic transformation of human epithelial cells. IARC Sci Publ. 1988. PMID: 3069724
- Brash DE, Mark GE, Farrell MP, Harris CC: Overview of human cells in genetic research: altered phenotypes in human cells caused by transferred genes. Somat Cell Mol Genet. 1987 Jul. PMID: 3331832
- Brash DE, Seetharam S, Kraemer KH, Seidman MM, Bredberg A: Photoproduct frequency is not the major determinant of UV base substitution hot spots or cold spots in human cells. Proc Natl Acad Sci U S A. 1987 Jun. PMID: 3473483
- Brash DE, Reddel RR, Quanrud M, Yang K, Farrell MP, Harris CC: Strontium phosphate transfection of human cells in primary culture: stable expression of the simian virus 40 large-T-antigen gene in primary human bronchial epithelial cells. Mol Cell Biol. 1987 May. PMID: 3037341
- Glickman BW, Schaaper RM, Haseltine WA, Dunn RL, Brash DE: The C-C (6-4) UV photoproduct is mutagenic in Escherichia coli. Proc Natl Acad Sci U S A. 1986 Sep. PMID: 3529093
- Sutherland BM, Oliveira OM, Ciarrocchi G, Brash DE, Haseltine WA, Lewis RJ, Hanawalt PC: Substrate range of the 40,000-dalton DNA-photoreactivating enzyme from Escherichia coli. Biochemistry. 1986 Feb 11. PMID: 3513832
- Brash DE, Franklin WA, Sancar GB, Sancar A, Haseltine WA: Escherichia coli DNA photolyase reverses cyclobutane pyrimidine dimers but not pyrimidine-pyrimidone (6-4) photoproducts. J Biol Chem. 1985 Sep 25. PMID: 3900062
- Brash DE, Haseltine WA: Photoreactivation of Escherichia coli reverses umuC induction by UV light. J Bacteriol. 1985 Aug. PMID: 2991189
- Su CM, Brash DE, Turturro A, Hart RW: Longevity-dependent organ-specific accumulation of DNA damage in two closely related murine species. Mech Ageing Dev. 1984 Oct 15. PMID: 6492898
- Su CM, Brash DE, Chang MJ, Hart RW, D'Ambrosio SM: Induction of single-strand breaks plus alkali-labile bonds by N-nitrosoureas in rat tissues in vivo: ethylnitrosourea versus benzylnitrosourea. Mutat Res. 1983 Mar. PMID: 6835219
- Brash DE, Haseltine WA: UV-induced mutation hotspots occur at DNA damage hotspots. Nature. 1982 Jul 8. PMID: 7045692
- Brash DE, Hart RW: Biopsy measurement of DNA damage and repair in vivo: single-strand breaks, alkali-labile bonds, and endonuclease-sensitive sites. Radiat Res. 1982 Jul. PMID: 6285409
- Lipetz PD, Brash DE, Joseph LB, Jewett HD, Lisle DR, Lantry LE, Hart RW, Stephens RE: Determination of DNA superhelicity and extremely low levels of DNA strand breaks in low numbers of nonradiolabeled cells by DNA-4',6-diamidino-2-phenylindole fluorescence in nucleoid gradients. Anal Biochem. 1982 Apr. PMID: 7103066
- Lippke JA, Gordon LK, Brash DE, Haseltine WA: Distribution of UV light-induced damage in a defined sequence of human DNA: detection of alkaline-sensitive lesions at pyrimidine nucleoside-cytidine sequences. Proc Natl Acad Sci U S A. 1981 Jun. PMID: 6943547
- Brash DE, Hart RW: DNA damage and repair in vivo. J Environ Pathol Toxicol. 1978 Sep-Oct. PMID: 363969
- Ziegler A, Jonason AS, Leffell DJ, Simon JA, Sharma HW, Kimmelman J, Remington L, Jacks T, Brash DE: Sunburn and p53 in the onset of skin cancer. Nature. 1994 Dec 22-29. PMID: 7997263