A novel germline MLH1 mutation causing Lynch Syndrome in patients from the Republic of Macedonia

Aim To implement molecular analysis in the clinical diagnosis and management of Lynch syndrome (LS). Methods We analyzed the mutations in MLH1 and MSH2 in the selected LS families from the Republic of Macedonia. Results We performed the very first genetic identification of LS families and characterized a novel mutation. The novel nonsense germline point mutation c.392C>G in the codon 131 of MLH1(S131X) was identified as the underlying genetic cause of LS in three families. The haplotype analysis suggested a founder effect of this mutation in our population. Conclusion We expect to detect the mutation in other LS patients from the region, and recommend costeffective screening for this mutation by restriction fragment length polymorphism-polymerase chain reaction or DNA sequencing of MLH1 Exon5 prior to full genetic testing in all LS suspects of Macedonian ancestry

Molecular profile of the Lynch Syndrome in the Republic of Macedonia

The most frequent type of hereditary colorectal cancer, the one occurring in the setting of the Lynch syndrome (LS) is considered a phenotypic manifestation of a germline defect in the mismatch repair mechanism i.e. in the MLH1, MSH2, MSH6 or PMS2 gene. Aiming towards establishment of a standardized protocol involving molecular analyses for diagnosis of this syndrome and developing a unique national register of families with hereditary colorectal cancer syndromes in the Republic of Macedonia, we began a prospective study to reveal the genetic defects among Macedonian patients with colorectal cancer (CRC) and identifying families with hereditary CRC. A total of 53 patients fulfilling the revised Bethesda criteria for MSI-genetic testing were compared to 350 patients with sporadic CRC. The results reveal significant differences in age at diagnosis (p=0.03), involvement of microsatellite instability (p<0.0001) and localization of the tumor in respect to flexura lienalis (p=0.009) and suggest affiliation of the majority of the “Bethesda+” CRCs to the so called Familial Colorectal cancer Type X group. The molecular characterization of LS suspects identified the novel MLH1 c.392C>G nonsense mutation with a possible founder effect in the Macedonian population, the MLH1 ex.3-12 deletion, as well as the c.244A>G mutation, IVS14- 19A>G and IVS4+65A>C changes in MLH1 without confirmed pathological significance. The observed high frequency (87.5%) of the Ile219Val (c.655A>G) variant in MLH1 among the LS suspects prompts further analyses to evaluate its involvement in the development of hereditary CRC by itself or as a risk modifying factor among the patients from the Republic of Macedonia