Dana-farber-cancer Institute Inc.

Other

Oregon Health & Science University

University of Vermont and State Agricultural College

The Brigham and Women's Hospital, Inc.

Albert Einstein College of Medicine

Richard D Kolodner

Roni J Bollag

C Eric Bronner

Robert M Liskay

Sean M Baker

Raju S Kucherlapati

Winfried Edelmann

Jeffrey W Pollard

Richard Fishel

Nena Winand

Robert A G Reenan

Jeffrey S Sklar

Robert Mashal

We have now discovered that eukaryotes, including mammals, have a DNA mismatch repair pathway analogous to the pathway that exists in bacteria. Defects or alterations in this mismatch repair pathway in a mammal, such as a human, will result in the accumulation of unstable repeated DNA sequences. Such a phenotype has a high correlation to disease state in a number of cancers, such as hereditary colon cancers. Accordingly, discovering a defect or alteration in the pathway can be diagnostic of a predisposition to cancer, and prognostic for a particular cancer. We have also discovered and sequenced one of the genes in this pathway in a number of mammals, including humans. This gene, referred to herein as MSH2, has many applications. It can be used in assays, to express gene product, for drug screens, and therapeutically. We also disclose herein a method for screening for other genes in this mismatch repair pathway.

Method for detection of alterations in the DNA mismatch repair pathway

We have now discovered that mammals have a DNA gene analogous to that existing in bacteria. MSH5 defects or alterations in this mismatch repair pathway in a mammal, such as a human, can be diagnostic of a predisposition to cancer, and prognostic for a particular cancer: We have discovered and sequenced MSH5 in a number of mammals, including humans. This gene can be used in assays, to express gene product, for drug screens, and therapeutically.

Method of detection of alterations in MSH5

We have now discovered that eukaryotes, including mammals, have a DNA mismatch repair pathway analogous to the pathway that exists in bacteria. Defects or alterations in this mismatch repair pathway in a mammal, such as a human, will result in the accumulation of unstable repeated DNA sequences. Such a phenotype has a high correlation to disease state in a number of cancers, such as hereditary colon cancers. Accordingly, discovering a defect or alteration in the pathway can be diagnostic of a predisposition to cancer, and prognostic for a particular cancer.

An animal, e.g., transgenic mouse, in which the MSH5 gene is misexpressed. The animal is useful for screening treatments for a number of conditions. Methods for identifying contraceptive agents are also described.

Methods for identifying compounds which modulate the activity of MSH5

Genomic sequences of human mismatch repair genes and gene products are described, as are methods of detecting mutations and/or polymorphisms in those genes. Also described are methods of diagnosing cancer susceptibility in a subject, and methods of identifying and classifying mismatch-repair-defective tumors. In particular, sequences and methods relating to human mutL homologs, hMLH1 and hPMS1 genes and gene products are provided.

Compositions and methods relating to DNA mismatch repair genes

MSH5 ablated mice and uses therefor

Genomic sequences of human mismatch repair genes are described, as are methods of detecting mutations and/or polymorphisms in those genes. Also described are methods of diagnosing cancer susceptibility in a subject, and methods of identifying and classifying mismatch-repair-defective tumors. In particular, sequences and methods relating to human mutL homologs, hMLH1 and hPMS1 genes are provided.

Composition and methods relating to DNA mismatch repair genes

We have discovered two human genes, hMLH1 and hPMS1, each of which apparently encodes for a protein involved in DNA mismatch repair. The hMLH1 gene encodes for a protein which is homologous to the bacterial DNA mismatch repair protein MutL, and is located on human chromosome 3p21.3-23. We believe that mutations in the hMLH1 gene cause hereditary non-polyposis colon cancer (HNPCC) in some individuals based upon the similarity of the hMLH1 gene product to the yeast DNA mismatch repair protein MLH1, the coincident location of the hMLH1 gene and the HNPCC locus on chromosome 3, and hMLH1 missense mutations in affected individuals from a chromosome 3-linked HNPCC family. The human hPMS1 gene is homologous to the yeast DNA mismatch repair gene PMS1, and is located on human chromosome 7q. We believe that the hPMS1 gene is a strong candidate for HNPCC testing because the yeast proteins MLH1 and PMS1 have been shown to be involved in the same DNA repair pathway and because hMLH1 and hMSH2 have both been implicated in HNPCC families. The most immediate use for hMLH1 and hPMS1 will be in screening tests on individuals who are members of families which exhibit high frequencies of early onset cancer. We have also isolated and sequenced mouse MLH1 and PMS1 genes. We have produced chimeric mice with a mutant form of the PMS1 gene that will enable us to derive mice that are heterozygous or homozygous for mutation in mPMS1. These mice will be useful for cancer research. We have also produced and isolated antibodies directed to hPMS1 which are useful in assays to detect the presence of protein in tumor samples.

Mammalian DNA mismatch repair genes MLH1 and PMS1

Novel methods and kits for detecting mismatches in nucleic acids using one or more resolvase enzymes. These methods can be used in medical procedures or research, veterinary procedures or research, agricultural applications, forensics, and paternity testing. For example, in forensics and paternity testing, methods of the invention can replace or complement RFLP mapping.

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Jamaica Plain, MA, United States of America

Richard D Kolodner