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1.
Genes Chromosomes Cancer ; 63(1): e23208, 2024 Jan.
Article in English | MEDLINE | ID: mdl-37795928

ABSTRACT

Polyketide synthase (pks) island harboring Escherichia coli are, under the right circumstances, able to produce the genotoxin colibactin. Colibactin is a risk factor for the development of colorectal cancer and associated with mutational signatures SBS88 and ID18. This study explores colibactin-associated mutational signatures in biallelic NTHL1 and MUTYH patients. Targeted Next Generation Sequencing (NGS) was performed on colorectal adenomas and carcinomas of one biallelic NTHL and 12 biallelic MUTYH patients. Additional fecal metagenomics and genome sequencing followed by mutational signature analysis was conducted for the NTHL1 patient. Targeted NGS of the NTHL1 patient showed somatic APC variants fitting SBS88 which was confirmed using WGS. Furthermore, fecal metagenomics revealed pks genes. Also, in 1 out of 11 MUTYH patient a somatic variant was detected fitting SBS88. This report shows that colibactin may influence development of colorectal neoplasms in predisposed patients.


Subject(s)
Adenomatous Polyposis Coli , Colorectal Neoplasms , Humans , Adenomatous Polyposis Coli/genetics , Adenomatous Polyposis Coli/pathology , Mutation , Colorectal Neoplasms/genetics , Colorectal Neoplasms/pathology , Deoxyribonuclease (Pyrimidine Dimer)/genetics
2.
Fam Cancer ; 17(3): 361-370, 2018 07.
Article in English | MEDLINE | ID: mdl-28933000

ABSTRACT

Until recently, no prediction models for Lynch syndrome (LS) had been validated for PMS2 mutation carriers. We aimed to evaluate MMRpredict and PREMM5 in a clinical cohort and for PMS2 mutation carriers specifically. In a retrospective, clinic-based cohort we calculated predictions for LS according to MMRpredict and PREMM5. The area under the operator receiving characteristic curve (AUC) was compared between MMRpredict and PREMM5 for LS patients in general and for different LS genes specifically. Of 734 index patients, 83 (11%) were diagnosed with LS; 23 MLH1, 17 MSH2, 31 MSH6 and 12 PMS2 mutation carriers. Both prediction models performed well for MLH1 and MSH2 (AUC 0.80 and 0.83 for PREMM5 and 0.79 for MMRpredict) and fair for MSH6 mutation carriers (0.69 for PREMM5 and 0.66 for MMRpredict). MMRpredict performed fair for PMS2 mutation carriers (AUC 0.72), while PREMM5 failed to discriminate PMS2 mutation carriers from non-mutation carriers (AUC 0.51). The only statistically significant difference between PMS2 mutation carriers and non-mutation carriers was proximal location of colorectal cancer (77 vs. 28%, p < 0.001). Adding location of colorectal cancer to PREMM5 considerably improved the models performance for PMS2 mutation carriers (AUC 0.77) and overall (AUC 0.81 vs. 0.72). We validated these results in an external cohort of 376 colorectal cancer patients, including 158 LS patients. MMRpredict and PREMM5 cannot adequately identify PMS2 mutation carriers. Adding location of colorectal cancer to PREMM5 may improve the performance of this model, which should be validated in larger cohorts.


Subject(s)
Colorectal Neoplasms, Hereditary Nonpolyposis/genetics , Heterozygote , Mismatch Repair Endonuclease PMS2/genetics , Models, Statistical , Adult , Area Under Curve , Cohort Studies , Female , Humans , Male , Middle Aged , ROC Curve , Retrospective Studies , Sensitivity and Specificity
3.
Clin Genet ; 93(1): 134-137, 2018 Jan.
Article in English | MEDLINE | ID: mdl-28503822

ABSTRACT

Constitutional mismatch repair deficiency (CMMRD) is a rare, recessively inherited childhood cancer predisposition syndrome caused by biallelic germline mutations in one of the mismatch repair genes. The CMMRD phenotype overlaps with that of neurofibromatosis type 1 (NF1), since many patients have multiple café-au-lait macules (CALM) and other NF1 signs, but no germline NF1 mutations. We report of a case of a healthy 6-year-old girl who fulfilled the diagnostic criteria of NF1 with >6 CALM and freckling. Since molecular genetic testing was unable to confirm the diagnosis of NF1 or Legius syndrome and the patient was a child of consanguineous parents, we suspected CMMRD and found a homozygous PMS2 mutation that impairs MMR function. Current guidelines advise testing for CMMRD only in cancer patients. However, this case illustrates that including CMMRD in the differential diagnosis in suspected sporadic NF1 without causative NF1 or SPRED1 mutations may facilitate identification of CMMRD prior to cancer development. We discuss the advantages and potential risks of this CMMRD testing scenario.


Subject(s)
DNA Mismatch Repair/genetics , DNA Repair Enzymes/deficiency , DNA Repair Enzymes/genetics , Child , Consanguinity , Family Health , Female , Humans , Mismatch Repair Endonuclease PMS2/genetics , Mutation , Pedigree
4.
Clin Genet ; 91(4): 536-544, 2017 Apr.
Article in English | MEDLINE | ID: mdl-27485256

ABSTRACT

Germline mutations in genes encoding subunits of succinate dehydrogenase (SDH) are associated with hereditary paraganglioma and pheochromocytoma. Although most mutations in SDHB, SDHC and SDHD are intraexonic variants, large germline deletions may represent up to 10% of all variants but are rarely characterized at the DNA sequence level. Additional phenotypic effects resulting from deletions that affect neighboring genes are also not understood. We performed multiplex ligation-dependent probe amplification, followed by a simple long-range PCR 'chromosome walking' protocol to characterize breakpoints in 20 SDHx-linked paraganglioma-pheochromocytoma patients. Breakpoints were confirmed by conventional PCR and Sanger sequencing. Heterozygous germline deletions of up to 104 kb in size were identified in SDHB, SDHC, SDHD and flanking genes in 20 paraganglioma-pheochromocytoma patients. The exact breakpoint could be determined in 16 paraganglioma-pheochromocytoma patients of which 15 were novel deletions. In six patients proximal genes were also deleted, including PADI2, MFAP2, ATP13A2 (PARK9), CFAP126, TIMM8B and C11orf57. These genes were either partially or completely deleted, but did not modify the phenotype. This study increases the number of known SDHx deletions by over 50% and demonstrates that a significant proportion of large gene deletions can be resolved at the nucleotide level using a simple and rapid method.


Subject(s)
Membrane Proteins/genetics , Paraganglioma/genetics , Succinate Dehydrogenase/genetics , Base Sequence/genetics , Chromosome Breakpoints , Female , Genetic Predisposition to Disease , Germ-Line Mutation , Heterozygote , Humans , Male , Paraganglioma/pathology , Sequence Deletion/genetics
5.
Fam Cancer ; 15(4): 563-70, 2016 10.
Article in English | MEDLINE | ID: mdl-26880076

ABSTRACT

Familial adenomatous polyposis (FAP) is a dominantly inherited syndrome caused by germline mutations in the APC gene and characterized by the development of multiple colorectal adenomas and a high risk of developing colorectal cancer (CRC). The severity of polyposis is correlated with the site of the APC mutation. However, there is also phenotypic variability within families with the same underlying APC mutation, suggesting that additional factors influence the severity of polyposis. Genome-wide association studies identified several single nucleotide polymorphisms (SNPs) that are associated with CRC. We assessed whether these SNPs are associated with polyp multiplicity in proven APC mutation carriers. Sixteen CRC-associated SNPs were analysed in a cohort of 419 APC germline mutation carriers from 182 families. Clinical data were retrieved from the Dutch Polyposis Registry. Allele frequencies of the SNPs were compared for patients with <100 colorectal adenomas versus patients with ≥100 adenomas, using generalized estimating equations with the APC genotype as a covariate. We found a trend of association of two of the tested SNPs with the ≥100 adenoma phenotype: the C alleles of rs16892766 at 8q23.3 (OR 1.71, 95 % CI 1.05-2.76, p = 0.03, dominant model) and rs3802842 at 11q23.1 (OR 1.51, 95 % CI 1.03-2.22, p = 0.04, dominant model). We identified two risk variants that are associated with a more severe phenotype in APC mutation carriers. These risk variants may partly explain the phenotypic variability in families with the same APC gene defect. Further studies with a larger sample size are recommended to evaluate and confirm the phenotypic effect of these SNPs in FAP.


Subject(s)
Adenomatous Polyposis Coli Protein/genetics , Chromosomes, Human, Pair 11 , Chromosomes, Human, Pair 8 , Colorectal Neoplasms/genetics , Adenoma/genetics , Adenomatous Polyposis Coli/genetics , Adult , Gene Frequency , Genetic Predisposition to Disease , Genome-Wide Association Study , Humans , Male , Mutation , Polymorphism, Single Nucleotide
6.
Fam Cancer ; 12(1): 43-50, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23007840

ABSTRACT

In the absence of a polyposis phenotype, colorectal cancer (CRC) patients referred for genetic testing because of early-onset disease and/or a positive family history, typically undergo testing for molecular signs of Lynch syndrome in their tumors. In the absence of these signs, DNA testing for germline mutations associated with other known tumor syndromes is usually not performed. However, a few studies in large series of CRC patients suggest that in a small percentage of CRC cases, bi-allelic MUTYH germline mutations can be found in the absence of the MUTYH-associated polyposis phenotype. This has not been studied in the Dutch population. Therefore, we analyzed the MUTYH gene for mutations in 89 patients with microsatellite-low or stable CRC cancer diagnosed before the age of 40 years or otherwise meeting the Bethesda criteria, all of them without a polyposis phenotype. In addition, we studied a series of 693 non-CRC patients with 1-13 adenomatous colorectal polyps for the MUTYH hotspot mutations Y179C, G396D and P405L. No bi-allelic MUTYH mutations were observed. Our data suggest that the contribution of bi-allelic MUTYH mutations to the development of CRC in Dutch non-polyposis patients that meet clinical genetic referral criteria, and to the development of low number of colorectal adenomas in non-CRC patients, is likely to be low.


Subject(s)
Adenomatous Polyps/genetics , Colorectal Neoplasms, Hereditary Nonpolyposis/genetics , Colorectal Neoplasms/genetics , DNA Glycosylases/genetics , Genetic Predisposition to Disease , Adult , Aged , Female , Germ-Line Mutation , Humans , Male , Middle Aged , Mutation , Netherlands , Young Adult
7.
Clin Genet ; 81(3): 284-8, 2012 Mar.
Article in English | MEDLINE | ID: mdl-21348866

ABSTRACT

Mutations in four genes encoding subunits or cofactors of succinate dehydrogenase (SDH) cause hereditary paraganglioma and pheochromocytoma syndromes. Mutations in SDHB and SDHD are generally the most common, whereas mutations in SDHC and SDHAF2 are far less frequently observed. A total of 1045 DNA samples from Dutch paraganglioma and pheochromocytoma patients and their relatives were analyzed for mutations of SDHB, SDHC, SDHD or SDHAF2. Mutations in these genes were identified in 690 cases, 239 of which were index cases. The vast majority of mutation carriers had a mutation in SDHD (87.1%). The second most commonly affected gene was SDHAF2 (6.7%). Mutations in SDHB were found in only 5.9% of samples, whereas SDHC mutations were found in 0.3% of samples. Remarkably, 69.1% of all carriers of a mutation in an SDH gene in the Netherlands can be attributed to a single founder mutation in SDHD, c.274G>T and p.Asp92Tyr. Moreover, 88.8% of all SDH mutation carriers carry one of just six Dutch founder mutations in SDHB, SDHD and SDHAF2. The dominance of SDHD mutations is unique to the Netherlands, contrasting with the higher prevalence of SDHB mutations found elsewhere. In addition, we found that most SDH mutation-related paragangliomas-pheochromocytomas in the Netherlands can be explained by only six founder mutations in SDHAF2, SDHB and SDHD. The findings underline the regional differences in the SDH mutation spectrum, differences that should be taken into account in the development of effective screening protocols. The results show the crucial role that demographic factors play in the frequency of gene mutations.


Subject(s)
Founder Effect , Mutation , Succinate Dehydrogenase/genetics , Adrenal Gland Neoplasms/genetics , Humans , Netherlands/epidemiology , Paraganglioma/genetics , Pheochromocytoma/genetics , Prevalence
8.
Clin Endocrinol (Oxf) ; 75(5): 650-5, 2011 Nov.
Article in English | MEDLINE | ID: mdl-21561462

ABSTRACT

OBJECTIVE: Head and neck paragangliomas (HNPGL) are associated with mutations in genes encoding subunits of succinate dehydrogenase (SDH). The aim of this study was to evaluate SDH mutations, family history and phenotypes of patients with HNPGL in the Netherlands. DESIGN: We evaluated the clinical data and the mutation status of 236 patients referred between 1950 and 2009 to Leiden University Medical Center. RESULTS: The large majority of the patients carried mutations in SDHD (83%), and the p.Asp92Tyr Dutch founder mutation in SDHD alone accounted for 72% of all patients with HNPGL. A mutation in SDHAF2 was found in 4%, mutations in SDHB in 3% and a mutation in SDHC was identified in a single patient (0·4%). Over 80% of patients presented with positive family history, of whom 99·5% carried a mutation in an SDH gene. SDH mutations were also found in 56% of isolated patients, chiefly in SDHD (46%), but also in SDHB (8%) and SDHC (2%). The clinical parameters of these different subgroups are discussed: including the age at diagnosis, associated pheochromocytomas, tumour multifocality and malignancy rate. CONCLUSION: The majority of Dutch patients with HNPGL present with a positive family history, in contrast to other European countries. The clinical characteristics of patients with HNPGL are chiefly determined by founder mutations in SDHD, the major causative gene in both familial and isolated patients with HNPGL. The high frequency of founder mutations in SDHD suggests a higher absolute prevalence of paraganglioma syndrome in the Netherlands.


Subject(s)
Head and Neck Neoplasms/genetics , Head and Neck Neoplasms/pathology , Paraganglioma/genetics , Paraganglioma/pathology , Succinate Dehydrogenase/genetics , Adult , Female , Humans , Male , Membrane Proteins/genetics , Middle Aged , Mutation , Netherlands
9.
Clin Genet ; 80(3): 243-55, 2011 Sep.
Article in English | MEDLINE | ID: mdl-21261604

ABSTRACT

Childhood brain tumours may be due to germline bi-allelic mismatch repair (MMR) gene mutations in MLH1, MSH2, MSH6 or PMS2. These mutations can also lead to colorectal neoplasia and haematological malignancies. Here, we review this syndrome and present siblings with early-onset rectal adenoma and papillary glioneural brain tumour, respectively, due to novel germline bi-allelic PMS2 mutations. Identification of MMR protein defects can lead to early diagnosis of this condition. In addition, assays for these defects may help to classify brain tumours for research protocols aimed at targeted therapies.


Subject(s)
Adenoma/genetics , Adenosine Triphosphatases , Brain Neoplasms/genetics , Colorectal Neoplasms/genetics , DNA Repair Enzymes , DNA-Binding Proteins , Germ-Line Mutation , Glioma/genetics , Adenoma/diagnosis , Adenoma/pathology , Adenosine Triphosphatases/genetics , Age of Onset , Brain Neoplasms/diagnosis , Brain Neoplasms/pathology , Child , Colorectal Neoplasms/diagnosis , Colorectal Neoplasms/pathology , DNA Mismatch Repair , DNA Mutational Analysis , DNA Repair Enzymes/genetics , DNA-Binding Proteins/genetics , Female , Glioma/diagnosis , Glioma/pathology , Heterozygote , Humans , Male , Microsatellite Repeats , Mismatch Repair Endonuclease PMS2 , Pedigree , Siblings , Syndrome , Young Adult
10.
Clin Genet ; 80(6): 558-65, 2011 Dec.
Article in English | MEDLINE | ID: mdl-21204794

ABSTRACT

Heterozygous germline mutations in the mismatch repair (MMR) genes MLH1, MSH2, MSH6 and PMS2 cause Lynch syndrome. Biallelic mutations in the MMR genes are associated with a childhood cancer syndrome [constitutional mismatch repair deficiency (CMMR-D)]. This is predominantly characterized by hematological malignancies and tumors of the bowel and brain, often associated with signs of neurofibromatosis type 1 (NF1). Diagnostic strategies for selection of patients for MMR gene analysis include analysis of microsatellite instability (MSI) and immunohistochemical (IHC) analysis of MMR proteins in tumor tissue. We report the clinical characterization and molecular analyses of tumor specimens from a family with biallelic PMS2 germline mutations. This illustrates the pitfalls of present molecular screening strategies. Tumor tissues of five family members were analyzed for MSI and IHC. MSI was observed in only one of the analyzed tissues. However, IHC analysis of brain tumor tissue of the index patient and his sister showed absence of PMS2 expression, and germline mutation analyses showed biallelic mutations in PMS2: p.Ser46IIe and p.Pro246fs. The same heterozygous mutations were confirmed in the father and mother, respectively. These data support the conclusion that in case of a clinical phenotype of CMMR-D, it is advisable to routinely combine MSI analysis with IHC analysis for the expression of MMR proteins. With inconclusive or conflicting results, germline mutation analysis of the MMR genes should be considered after thorough counselling of the patients and/or their relatives.


Subject(s)
Adenosine Triphosphatases/genetics , DNA Repair Enzymes/genetics , DNA-Binding Proteins/genetics , Gene Expression Regulation, Neoplastic , Germ-Line Mutation , Adult , Aged, 80 and over , Brain Neoplasms/diagnosis , Brain Neoplasms/genetics , Brain Neoplasms/pathology , Child , DNA Mismatch Repair , DNA Repair-Deficiency Disorders/genetics , Female , Genetic Testing , Heterozygote , Humans , Immunohistochemistry , Male , Microsatellite Instability , Middle Aged , Mismatch Repair Endonuclease PMS2 , MutS Homolog 2 Protein/genetics , Neoplastic Syndromes, Hereditary/diagnosis , Neoplastic Syndromes, Hereditary/genetics , Pedigree
11.
Ophthalmic Genet ; 31(4): 205-8, 2010 Dec.
Article in English | MEDLINE | ID: mdl-21067481

ABSTRACT

Peters plus syndrome is an autosomal recessive rare congenital disorder defined by corneal Peters anomaly with short disproportionate stature, development delay and dysmorphic facial features. In addition, cardiac, genito-urinary and/or central nervous system malformations can be present. Mutations in the beta-1,3-galactosyltransferase-like glycosyltransferase gene (B3GALTL) have been reported in patients with Peters plus syndrome prompting phenotype-genotype studies because of the variable clinical spectrum related to the syndrome. A 20 month old boy presenting with bilateral Peters anomaly in association with multiple developmental anomalies including cerebral malformations was found to carry a novel homozygous B3GALTL nonsense mutation [p.Tyr366X]. This is the first stop mutation described in association with this gene. The present report confirms the wide clinical spectrum of Peters plus syndrome, underlines the major clinical criteria of the syndrome and the major implication of B3GALTL gene in this condition. Ophthalmologic examination in multiple developmental anomalies remains an important clinical issue that may lead to specific gene screening. In Peters plus syndrome B3GALTL molecular test provides diagnosis confirmation and improves dramatically genetic counselling for the families.


Subject(s)
Abnormalities, Multiple/genetics , Codon, Nonsense , Galactosyltransferases/genetics , Glucosyltransferases/genetics , Anterior Eye Segment/abnormalities , Corneal Opacity/genetics , Developmental Disabilities/genetics , Eye Abnormalities/genetics , Humans , Infant , Male , Nervous System Malformations/genetics , Polymerase Chain Reaction , Syndrome
12.
J Pathol ; 216(1): 25-31, 2008 Sep.
Article in English | MEDLINE | ID: mdl-18506705

ABSTRACT

Genetic instability is known to drive colorectal carcinogenesis. Generally, a distinction is made between two types of genetic instability: chromosomal instability (CIN) and microsatellite instability (MIN or MSI). Most CIN tumours are aneuploid, whereas MSI tumours are considered near-diploid. However, for MUTYH-associated polyposis (MAP) the genetic instability involved in the carcinogenesis remains unclear, as near-diploid adenomas, aneuploid adenomas and near-diploid carcinomas have been reported. Remarkably, our analysis of 26 MAP carcinomas, using SNP arrays and flow sorting, showed that these tumours are often near-diploid (52%) and mainly contain chromosomal regions of copy-neutral loss of heterozygosity (LOH) (71%). This is in contrast to sporadic colon cancer, where physical loss is the main characteristic. The percentage of chromosomal gains (24%) is comparable to sporadic colorectal cancers with CIN. Furthermore, we verified our scoring of copy-neutral LOH versus physical loss in MAP carcinomas by two methods: fluorescence in situ hybridization, and LOH analysis using polymorphic markers on carcinoma fractions purified by flow sorting. The results presented in this study suggest that copy-neutral LOH is an important mechanism in the tumorigenesis of MAP.


Subject(s)
Adenomatous Polyposis Coli/genetics , Chromosomal Instability/genetics , DNA Glycosylases/genetics , Loss of Heterozygosity/genetics , Adenomatous Polyposis Coli/pathology , Adult , Aged , Biomarkers, Tumor/genetics , DNA Mutational Analysis/methods , Humans , Microsatellite Instability , Middle Aged
13.
Gut ; 57(1): 71-6, 2008 Jan.
Article in English | MEDLINE | ID: mdl-17604324

ABSTRACT

BACKGROUND: The patient with 10 or more adenomas in the colon poses a diagnostic challenge. Beside germline mutations in the APC and MUTYH genes, only four cases of mosaic APC mutations have been reported. AIM: Given the relatively high frequency of de novo APC mutations in familial adenomatous polyposis (FAP), an investigation was carried out into whether the proportion of somatic mosaic APC mutations is currently underestimated. METHODS: Between 1 January 1994 and 31 December 2005 germline mutation analysis was performed in 599 consecutive index patients with polyposis coli referred for diagnostic APC scanning using a combination of denaturing gradient gel electrophoresis (DGGE) and protein truncation test (PTT). Variants were analysed by direct sequencing with primers flanking those used for DGGE and PTT, and quantified using pyrosequencing. RESULTS: Scrutinizing the molecular genetic results and family data of 242 index patients with pathogenic APC mutations led to the identification of 10 mosaic cases (4%). C>T transitions were observed in CGA sites in four of the 10 cases with somatic mosaicism, which is significantly more than 26 of the 232 non-mosaic cases (p = 0.02). Phenotypes of patients with somatic mosaicism ranged from an attenuated form of polyposis coli to florid polyposis with major extracolonic manifestations. CONCLUSIONS: Mosaicism occurs in a significant number of APC mutations and it is estimated that one-fifth of the de novo cases of FAP are mosaic. Clinically, the severity of manifestations in offspring and the recurrence risk for siblings of apparently sporadic polyposis patients may be underestimated due to parental APC mosaicism.


Subject(s)
Adenomatous Polyposis Coli/genetics , Genes, APC , Mosaicism , Adult , Aged , Cohort Studies , Female , Genotype , Germ-Line Mutation , Humans , Male , Middle Aged , Pedigree , Phenotype
14.
Clin Genet ; 72(6): 562-7, 2007 Dec.
Article in English | MEDLINE | ID: mdl-17922849

ABSTRACT

This study evaluated the use of genetic testing and time trends in hereditary non-polyposis colorectal cancer (HNPCC), (attenuated) familial adenomatous polyposis [(A)FAP] and human MutY homolog (MUTYH) associated polyposis (MAP) families. Eighty-seven families, who were diagnosed with disease-causing mutations between 1995 and 2006, were included in this study. The families consisted of 1547 individuals at risk. Data of these individuals were collected from medical records and family pedigrees. There was considerable interest in genetic testing with test rates of 41% in HNPCC families, 42% in (A)FAP families and 53% in MAP families. The use of genetic testing was associated with age and parenthood. Despite the interest in genetic testing, many risk carriers do not apply for testing. Moreover, time trend analysis showed a decline in test rate in HNPCC families. Studies evaluating the reasons for not testing are needed. Furthermore, a better implementation of genetic testing in clinical practice is desirable.


Subject(s)
Colorectal Neoplasms/genetics , Adenomatous Polyposis Coli/genetics , Cohort Studies , Colorectal Neoplasms, Hereditary Nonpolyposis/genetics , DNA Glycosylases/genetics , DNA Mutational Analysis , Female , Genetic Testing , Humans , Male , Risk Factors , Syndrome
15.
Clin Genet ; 71(5): 427-33, 2007 May.
Article in English | MEDLINE | ID: mdl-17489848

ABSTRACT

A small fraction of families with familial adenomatous polyposis (FAP) display an attenuated form of FAP (AFAP). We aimed to assess the presence of germline mutations in the MUTYH and adenomatous polyposis coli (APC) genes in AFAP families and to compare the clinical features between the two causative genes. Families with clinical AFAP were selected from the Dutch Polyposis Registry according to the following criteria: (a) at least two patients with 10-99 adenomas diagnosed at age >30 years or (b) one patient with 10-99 adenomas at age >30 years and a first-degree relative with colorectal cancer (CRC) with a few adenomas, and, applying for both criteria, no family members with more than 100 polyps before the age of 30 years. All probands were screened for germline mutations in the APC and MUTYH genes. Twenty-five of 315 Dutch families with FAP (8%) met our criteria for AFAP. These families included 146 patients with adenomas and/or CRC. Germline APC mutations were identified in nine families and biallelic MUTYH mutations in another nine families. CRC was identified at a mean age of 54 years (range 24-83 years) in families with APC and at 50 years (range 39-70 years) in families with MUTYH (p = 0.29). APC and biallelic MUTYH mutations are responsible for the majority of AFAP families. Based on our results and those reported in the literature, we recommend colonoscopy once every 2 years in AFAP families, starting surveillance from the late teens in APC mutation carriers and from age 20-25 years in biallelic MUTYH mutation carriers.


Subject(s)
Adenomatous Polyposis Coli/genetics , DNA Glycosylases/genetics , Genes, APC , Germ-Line Mutation , Adenomatous Polyposis Coli/diagnosis , Adult , Aged , Aged, 80 and over , Colorectal Neoplasms/genetics , DNA Mutational Analysis , Genetic Predisposition to Disease , Humans , Middle Aged , Mutation , Netherlands , Registries
16.
J Med Genet ; 42(9): e54, 2005 Sep.
Article in English | MEDLINE | ID: mdl-16140997

ABSTRACT

OBJECTIVE: To investigate the contribution of MYH associated polyposis coli (MAP) among polyposis families in the Netherlands, and the prevalence of colonic and extracolonic manifestations in MAP patients. METHODS: 170 patients with polyposis coli, who previously tested negative for APC mutations, were screened by denaturing gradient gel electrophoresis and direct sequencing to identify MYH germline mutations. RESULTS: Homozygous and compound heterozygous MYH mutations were identified in 40 patients (24%). No difference was found in the percentage of biallelic mutation carriers between patients with 10-99 polyps or 100-1000 polyps (29% in both groups). Colorectal cancer was found in 26 of the 40 patients with MAP (65%) within the age range 21 to 67 years (median 45). Complete endoscopic reports were available for 16 MAP patients and revealed five cases with gastro-duodenal polyps (31%), one of whom also presented with a duodenal carcinoma. Breast cancer occurred in 18% of female MAP patients, significantly more than expected from national statistics (standardised morbidity ratio = 3.75). CONCLUSIONS: Polyp numbers in MAP patients were equally associated with the attenuated and classical polyposis coli phenotypes. Two thirds of the MAP patients had colorectal cancer, 95% of whom were older than 35 years, and one third of a subset of patients had upper gastrointestinal lesions. Endoscopic screening of the whole intestine should be carried out every two years for all MAP patients, starting from age 25-30 years. The frequent occurrence of additional extraintestinal manifestations, such as breast cancer among female MAP patients, should be thoroughly investigated.


Subject(s)
Adenomatous Polyposis Coli/genetics , DNA Glycosylases/genetics , Adolescent , Adult , Aged , Child , Colorectal Neoplasms/genetics , DNA Mutational Analysis , Female , Genetic Predisposition to Disease , Genotype , Germ-Line Mutation , Humans , Inheritance Patterns/genetics , Male , Middle Aged , Netherlands , Phenotype , Risk
17.
Dis Markers ; 20(4-5): 207-13, 2004.
Article in English | MEDLINE | ID: mdl-15528786

ABSTRACT

Hereditary nonpolyposis colorectal cancer (HNPCC, Lynch syndrome) is a dominantly inherited syndrome characterized by the development of colorectal cancer, endometrial cancer and other cancers and the presence of microsatellite instability (MSI) in tumors. The Bethesda guidelines have been proposed for the identification of families suspected of HNPCC that require further molecular analysis. We have evaluated the yield of MSI-analysis in a large series of Dutch families suspected of HNPCC. We also analysed whether the loss of mismatch repair (MMR) protein detected by immunohistochemistry (IHC) of colorectal cancer (CRC) and endometrial cancer correlated with the presence of MSI and/or a MMR gene mutation. The results showed that the Bethesda criteria with a few modifications are appropriate to identify families eligible for genetic testing. In addition, we found that MSI and IHC-analysis of CRC using antibodies against MLH1, MSH2, MSH6 and PMS2 proteins are equally effective for identifying carriers of the known MMR gene defects. However, as long as the role of other putative MMR genes in hereditary CRC has not been elucidated, IHC-analysis cannot completely replace MSI. For this reason, we prefer MSI-analysis as first step in families suspected of HNPCC. On the other hand, in families fulfilling the revised Amsterdam criteria in which the probability of detecting a mutation is relatively high, we would recommend IHC as first diagnostic step because the result might predict the specific underlying MMR gene mutation. MSI or IHC-analysis of endometrial cancer alone was found to be less sensitive compared with these tests performed in colorectal cancer. Therefore, probably the best approach in the analysis of this cancer is to perform both techniques. The identification of HNPCC is important as it makes it possible to target effective preventative measures. Our studies showed that MSI and IHC analysis of colorectal and endometrial cancer, are reliable cost-effective tools that can be used to identify patients with HNPCC.


Subject(s)
Biomarkers, Tumor , Colorectal Neoplasms, Hereditary Nonpolyposis/diagnosis , Colorectal Neoplasms, Hereditary Nonpolyposis/genetics , Colorectal Neoplasms/diagnosis , Colorectal Neoplasms/genetics , Endometrial Neoplasms/diagnosis , Endometrial Neoplasms/genetics , Microsatellite Repeats , Adult , Colorectal Neoplasms/metabolism , Colorectal Neoplasms, Hereditary Nonpolyposis/metabolism , DNA Mutational Analysis , DNA Sequence, Unstable , Endometrial Neoplasms/metabolism , Female , Humans , Immunohistochemistry , Male , Middle Aged , Mutation , Phenotype , Syndrome
18.
Cas Lek Cesk ; 143(8): 543-6, 2004.
Article in Czech | MEDLINE | ID: mdl-15446460

ABSTRACT

BACKGROUND: Protein Truncation Test (PTT) was used to detect mutations in exon 15 of the APC gene in patients with Familial Adenomatous Polyposis. This method is limited by its ability to detect polypetide chains up to a certain minimum length. The aim of this study was to increase the sensitivity of detection of mutations in this region by using the technique of Denaturing Gradient Gel Electrophoresis (DGGE). METHODS AND RESULTS: Study were performed on 122 patients without detected mutations in the APC gene. The patients were divided into two independent groups 15A and 15A+15B (with 51 and 71 patients respectively). All the patients were tested with the DGGE and the positive findings were confirmed with sequencing. No mutation was detected in the group 15A (0%). In group 15A+15B one (1.4%) polymorphism and four (5.63%) patients with nonsense mutations were detected. CONCLUSIONS: DGGE is an effective method for detecting mutations in the first part of exon 15 of APC gene. It allows detecting any change in DNA strand. DGGE complements PTT in scanning of the whole exon 15 of APC gene.


Subject(s)
Adenomatous Polyposis Coli/genetics , Genes, APC , Mutation , Electrophoresis , Humans , Polymerase Chain Reaction
20.
Oncogene ; 20(43): 6241-4, 2001 Sep 27.
Article in English | MEDLINE | ID: mdl-11593433

ABSTRACT

Recently, we and others reported instability in the (C)8 repeat in exon 5 of MSH6 as a preferential target for somatic mutations in tumours from MSH6 germline mutation carriers. Here, we report that in 45% of tumours from MLH1, MSH2 and MSH6 germline mutation carriers no sequence change in the (C)8 repeat of MSH6 was found upon DNA sequencing analysis of PCR products with a shift in electrophoresis mobility. Using "standard" PCR primers a high frequency of instability (50-86%) of the (C)8 repeat was found, but using a modified PCR reverse primer, accomplishing modulation of non-templated addition of adenine during in vitro PCR amplification by the Taq polymerase, a markedly lower frequency of instability was found in tumours from MLH1, MSH2 and MSH6 mutation carriers (6, 13 and 40%, respectively). Furthermore, a significant difference of the frequency of instability of the (C)8 repeat in tumours from MSH6 mutation carriers was found compared to MLH1, MSH2 mutation carriers. These results might have important implications for the detection of instability of other short mononucleotide repeats, e.g. TGFbetaRII, BAX, IGFRII, PTEN, BRCA2.


Subject(s)
DNA-Binding Proteins , Fungal Proteins/genetics , Saccharomyces cerevisiae Proteins , Trinucleotide Repeat Expansion , Adaptor Proteins, Signal Transducing , Bias , Carrier Proteins , DNA Mutational Analysis , DNA Primers/metabolism , Exons , Gene Deletion , Humans , Microsatellite Repeats , MutL Protein Homolog 1 , MutS Homolog 2 Protein , Mutation , Neoplasm Proteins/genetics , Nuclear Proteins , Polymerase Chain Reaction , Proto-Oncogene Proteins/genetics , Reproducibility of Results , Sensitivity and Specificity
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