Men1 mutations that disrupt the coding sequence or splice sites can be detected in 80% to 90% of familial men1, and in approximately 65% of simplex men1 patients without a family history for the disease. The american college of medical genetics and genomics has published recommendations for reporting incidental findings in the exons of certain genes. Nov 15, 2016 dna was extracted from peripheral blood. Global gene expression in neuroendocrine tumors from. The amino acid sequence of this putative tumor suppressor offers no clue to the function or subcellular location of the protein. A men1 syndrome with a paraganglioma european journal of. Given a sample of dna, a dna sequencer is used to determine the order of the four bases. Multiple endocrine neoplasia type 1 men1 is diagnosed when two out of the three primary men1associated endocrine tumors occur in a patient. Brooks life sciences services tools for viewing sanger sequencing data sequence chromatogram viewing software. The first automated dna sequencer, invented by lloyd m. Rapid mutation screening for hrpt2 and men1 mutations. See structural alignment software for structural alignment of proteins. The human men1 gene spans 9 kb of genomic dna and is organised into 10 exons, whose sizes are indicated, and nine introns. Generation of two men1 knockout lines from a human embryonic.
In addition, mlpa was used for aip to detect large deletions or duplications. Clinical and genetic analysis of multiple endocrine. Clinical and genetic analysis of multiple endocrine neoplasia. Insulinomas, which are rare tumors causing hyperinsulinemic hypoglycemia are usually sporadic but may also occur in association with multiple endocrine neoplasia type 1 men 1. It is unclear if the phenotype and course of the disease of mutationnegative patients is comparable with mutationpositive patients and if these patients have true men1. Discriminant analysis using the scattergraph function of navigator software of exon 9 wild type, d418d c. Insulinomas, which are rare tumors causing hyperinsulinemic hypoglycemia are usually sporadic but may also occur in association with multiple endocrine neoplasia type 1 men 1 syndrome an autosomal dominant disorder caused by men1 gene mutations. Its most common endocrine expressions are tumors of parathyroids, enteropancreatic neuroendocrine tissue, and anterior pituitary. Sanger sequencing and fragment analysis software thermo. These sequences represent the protein coding region of the men1 cdna orf. Apr 27, 2017 wholeexome sequencing of the men1 gene of a patient with men1. Its most common endocrine expressions are tumors of parathyroids, enteropancreatic neuro.
Wes revealed two somatic functional variants outside. Men1 is inherited as an autosomal dominant disease, with a 50% risk of carrying the defective gene in firstdegree relatives. Men1 gene using wes, which was confirmed by sanger sequencing. Tools for viewing sequencing data resources genewiz.
Pdf characterization of the mouse men1 gene and its. An overview of detected somatic mutations after exome sequencing is provided in table 3. Primary tumors may be found in more than one endocrine organ andor multiple tumors may be found in the same organ. Menin, the product of the men1 gene, is a nuclear protein. The presence of a men1 mutation was determined with dna sequencing since 1998 and with a combination of dna sequencing and. Clinvar and hgmd into our clinical interpretation software to make the. Glu116 mutation was found in exon 2 of the men1 gene in ins2. The process simultaneously identifies dna bases while incorporating them into a nucleic acid chain. Men1 encodes a nuclear protein menin, a tumor suppressor which acts as an adapter and interacts with partner proteins involved in crucial activities. Objective multiple endocrine neoplasia type 1related primary hyperparathyroidism mhpt differs in many aspects from sporadic phpt shpt. Your search for men1 returned 110 pcrsanger sequencing primer pairs. To examine the men1 gene alterations in this family, blood samples were drawn from the affected figure 1, ii1 and 3 and asymptomatic members figure 1, ii2. Before starting new experimental designs or doing illumina sequencing for the first time, it is often helpful to look at the results of similar studies.
Recently, collagenomas and multiple angiofibromas of the dermis also have been recognized as very common. A heterozygous germline inactivating mutation in the men1 gene first hit may be followed by somatic loss of the remaining normal copy or somatic mutations in the men1 gene. Nov 15, 2016 direct sequencing of the gross deletion mutation of the men1 gene in family 3. Sanger sequencing dna sequencing software sequencher from. This list of sequence alignment software is a compilation of software tools and web portals used in pairwise sequence alignment and multiple sequence alignment. Pdf germline mutation landscape of multiple endocrine. The aims of this study were to summarize the clinical features and genetic background of chinese mhpt patients and compare the severity of the disease with those of shpt.
Novel association of men1 gene mutations with parathyroid. The men1 gene was screened, revealing three variants in cis at the intronexon 3 boundary ivs23gc, c. Lossoffunction germline men1 gene mutations account for 7595% of patients with multiple endocrine neoplasia type 1 men1. It used the sanger sequencing method, a technology which formed the basis of the first generation of dna sequencers and enabled the completion of the human genome project in 2001. Novel men 1 gene findings in rare sporadic insulinomaa. Acmg recommendations for reporting of incidental findings in clinical exome and genome sequencing. Sequences were analyzed with sequencing analysis software version 5. Analyze dna sequencing data from large or small whole genomes, whole exomes, targeted gene regions, and more with our userfriendly tools. Blueprint genetics men1 single gene test men1 single gene test. Men1 gene nonsense mutations occurred in two different tumor samples. In addition, the single gene test includes noncoding deep intronic variants if a tab for noncoding variants is displayed above. Microbebridge software is a streamlined, desktop software solution that connects dna sequences generated on applied biosystems sanger sequencers with the centers for disease control and prevention cdcs microbenet database for bacterial identification using 16s rrna gene sequencing analysis. Familial hyperparathyroidism, a disease of the parathyroid glands, may occur in conjunction with pituitary and pancreatic tumors multiple endocrine neoplasia type i, kidney and bone tumors hyperparathyroidism jaw tumor syndrome, or alone familial isolated hyperparathyroidism. It includes any method or technology that is used to determine the order of the four bases.
Multiple endocrine neoplasia type 1 men1 is a promising model to understand endocrine and other tumors. Schematic representation of the genomic structure and position of primers in the men1 gene scg2, menin. It has been postulated that mutations in noncoding. Wes and the snp array revealed somatic loh on chromosome 11 in parathyroid tumors left upper, left lower, and right upper parathyroid. Here, we generated two men1 knockout human embryonic stem cell lines, wae001a4 and wae001a5, by targeting exon2 and exon9 of men1 using the. Direct sequencing of pcr products was performed using a taqbig dye terminator sequencing kit and an abi3730 automated sequencer. Germline and somatic genetic changes in multicentric tumors. Fast, highquality, sampletodata services such as rna and wholegenome sequencing. A heterozygous germline inactivating mutation in the men1 gene first hit may be followed by somatic loss of the remaining normal copy or somatic mutations in the men1 gene second hit. Men1 gene cdna orf clone, homo sapienshuman genscript.
Wholeexome sequencing has been successfully used to elucidate the mutations associated with the different types of tumors. However, a higher aggressiveness of men1associated gastroenteropancreatic gep neuroendocrine tumours nets tumours has been reported when men1 gene truncating mutations are detected. Acmg recommendations for reporting of incidental findings in. Multiple endocrine neoplasia type 1 men1 sequencing. Illumina nextgeneration sequencing ngs technology uses clonal amplification and sequencing by synthesis sbs chemistry to enable rapid, accurate sequencing. Design and methods a total of 40 mhpt 27 sporadic, 7 families and 169 shpt cases of. The clinical analysis adhered to the established guidelines. We offer a wide range of nextgeneration sequencing ngs data analysis software tools, including pushbutton tools for dna sequence alignment, variant calling, and data visualization. Tyr227 mutation was found in exon 4 of the men1 gene in ins1, and c. A number of free software programs are available for viewing trace or chromatogram files. Nov 15, 2016 multiple endocrine neoplasia type 1 men1 is diagnosed when two out of the three primary men1associated endocrine tumors occur in a patient. Wes and the snp array revealed somatic loh on chromosome 11 in parathyroid tumors left upper, left lower, and right upper.
We found a novel germline truncating mutation of men1 gene at exon 10 in a subject with an aggressive clinical behavior of gepnets. The disease is caused by the functional loss of the tumor suppressor protein menin, coded by the men1 gene. To examine the men1 gene alterations in this family, blood samples were drawn from the affected figure 1, ii1 and 3 and asymptomatic members. This study describes the development and validation of rapid scanning for mutations in two tumor suppressor genes. However, a higher aggressiveness of men1 associated gastroenteropancreatic gep neuroendocrine tumours nets tumours has been reported when men1 gene truncating mutations are detected. The gene is located on chromosome 11q and encodes a multifunctional nuclear scaffold protein menin with a known role in the regulation of transcription, proliferation, differentiation, and genomic integrity. Generation of two men1 knockout lines from a human. Men1 redefined, a clinical comparison of mutationpositive. Two nonsense somatic mutations in men1 identified in.
Dna sequencing data analysis simple software tools. Rna sequencing revealed that the ivs23cg variant caused the skipping of the exon 3. These sequences represent the protein coding region of the men1 cdna orf which is encoded by the open reading frame orf sequence. A in order to find the exact deletion site of the gross deletion mutation detected by mlpa men1 del exon 5, 6, longrange pcr was used to amplify the region spanning exons 47 of the men1 gene. Gene codes will send an email update when the new, fully compatible version is released. Multiple endocrine neoplasia type 1 men1, omim 1100 is an autosomal dominant disorder characterized by endocrine tumors of the parathyroids, pancreatic islets and pituitary. Global gene expression in neuroendocrine tumors from patients. Smith, was introduced by applied biosystems in 1987.
Targeted yield plasmid dna prep glpcompliant services confirmatory sequencing snp mutation analysis. A somatic mutation in men1 gene detected in periventricular. Acmg recommendations for reporting of incidental findings. Wholeexome sequencing of the men1 gene of a patient with men1. The information provided here is not sufficient for interface builds. Multiple endocrine neoplasia type 1 men1 is characterized by endocrine tumors, particularly in the parathyroid glands, anterior pituitary, and pancreatic islet cells. Bwa, velvet, maq, gsnap and tablet are only the start for sequencher. The following men1 gene cdna orf clone sequences were retrieved from the ncbi reference sequence database refseq. The human men1 gene spans 9 kb of genomic dna and is organised into 10. Germline and somatic genetic changes in multicentric. Characterization of the mouse men1 gene and its expression during development article pdf available in oncogene 1719. However, we did not detect a somatic men1 gene mutation or loh in the pancreatic insulinoma. Novel men 1 gene findings in rare sporadic insulinomaa case. Mlpa mrcholland, amsterdam, the netherlands is used for detecting large deletions or duplications in the men1 gene.
This can help guide workflow modifications and select appropriate analytical software tools. The most recent version recommendation is acmg sf v2. Two nonsense somatic mutations in men1 identified in sporadic. A dna sequencer is a scientific instrument used to automate the dna sequencing process. Dna sequencing is the process of determining the nucleic acid sequence the order of nucleotides in dna. The american college of medical genetics and genomics has published recommendations for.
The presence of a men1 mutation was determined with dna sequencing since 1998 and with a combination of dna sequencing and multiplex ligationdependent probe amplification mlpa since 2005. Men type 1 men1 sequencing specimen component test codes cannot be used to order tests. Dna was extracted from fresh parathyroid tissue and peripheral blood with a maxwell 16 instrument, according to the manufacturers instructions promega corp, madison, wisconsin. The diagnosis of multiple endocrine neoplasia type 1 men1 syndrome should be suspected in individuals with endocrine tumors, although nonendocrine tumors may appear before the manifestations of hormonesecreting endocrine tumors see clinical description. For the men1 gene, 100% of the coding region was covered at a minimum of 10 the clinical analysis adhered to the established guidelines. Genetic sequencing is the future of medicine the washington. The men1 gene, mutations in which are responsible for multiple endocrine neoplasia type 1 men1, encodes a 610amino acid protein, denoted menin. The aims of this study were to summarize the clinical. This can help guide workflow modifications and select appropriate analytical software. An individual with a personal andor family history of tumors associated with multiple endocrine neoplasia, type i men1 especially parathyroid tumors, gastroenteropancreatic neuroendocrine tumors, and anterior pituitary tumors. Genetic analysis of parathyroid and pancreatic tumors in a. We can write dna software, boot it up to a computer converter and create unlimited variations of the gene sequences of biological life.
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