Project Summary
Lynch syndrome (LS), the most common hereditary cancer condition, is usually caused by a heritable genetic
mutation affecting one of the DNA mismatch repair genes, most frequently MLH1, located on chromosome 3.
So-called “MLH1 epimutation” is an alternative cause for LS. MLH1 epimutations are characterized by
methylation of a single copy of the MLH1 promoter, accompanied by loss of gene activity, throughout normal
tissues. This predisposes carriers to the development of LS-type cancers. Patients are identified by the
detection of “constitutional” MLH1 methylation in DNA from blood or any other normal tissue. Studies of the
family members of cases with an MLH1 epimutation have revealed distinct patterns of inheritance and hence
differing risks of passing the epimutation from parent to offspring, if at all. Therefore, MLH1 epimutations
provide a unique opportunity in humans for studying how epigenetic alterations arise and thereby cause
disease, and also how epigenetic alterations are transmitted from one generation to the next in some families,
but not in others. Dr. Hitchins, the principle investigator of this study, has assembled a unique collection of
families with an MLH1 epimutation as the cause for cancer, and has established the inheritance patterns of the
epimutation in each family, through her work in this field over the past 13 years. The goals of this study are to
determine the mechanisms that underlie MLH1 epimutation by undertaking an in-depth study of the genomes
and epigenomes of the patients and their families. In Aim 1, we will use state of the art “linked-read whole
genome sequencing” to sequence the entire genome of selected patients and family members to determine if
various types of genetic mutations on chromosome 3, or other chromosomes, underlie the onset on MLH1
epimutations. In Aim 2, we will determine if MLH1 is the only gene that is subject to erroneous constitutional
methylation, or if other genes are simultaneously methylated. This would provide insights into whether
epimutations arise because of a generalized epigenetic perturbation, or a focal mechanism affecting MLH1
alone. In Aim 3, we will validate any genetic or epigenetic alterations identified and determine their segregation
patterns in families. We will also perform functional analyses of genetic variants identified previously and in
Aim 1. Collectively, these studies will provide evidence for a causal link between genetic alterations identified
and the MLH1 epimutation, and enable us to predict the likelihood of inheritance between generations. Finally,
we will study patient white blood cells induced into a pluripotent state to model the epigenetic reprogramming
events that occur at MLH1 in these patients. This will shed light on why some epimutations are inherited and
others not. If successful, this study will enable us to improve the clinical management and genetic counselling
of families with a cancer-predisposing MLH1 epimutation. More broadly, this study will provide significant new
insights into the interaction between disease-causing genetic and epigenetic states, as well how altered
epigenetic states may are passed from one generation to the next, or erased between generations.