POSITIONAL CLONING OF THE MEN1 GENE

MEN1 基因的定位克隆

• 批准号: 7734860
• 负责人: settara chandrasekharappa
• 金额: $ 116.71万
• 依托单位: NATIONAL HUMAN GENOME RESEARCH INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7734860
• 关键词: 2q37; Affect; Amino Acid Sequence; Anterior Pituitary Gland; Antigens; Autistic Disorder; Biological Process; Breast; Breast Cancer Cell; Breast Cancer Model; CD15 Antigens; Cancer cell line; Cataloging; Catalogs; Cell Adhesion Molecules; Cell Differentiation process; Cell Line; Cell Nucleus; Cells; Cellular biology; Characteristics; Chondroitin Sulfate Proteoglycan; Clinical; Colon Carcinoma; Complex; Cytokeratin; DNA Damage; Defect; Development; Developmental Delay Disorders; Disease; Embryo; Endocrine; Endoderm; Enzymes; Evaluation; Face; Family; Fibroblasts; Gene Mutation; Gene Targeting; Genes; Genomics; Genotoxic Stress; Germ-Line Mutation; Hand; Heart; Hematopoiesis; Hematopoietic; Hereditary Malignant Neoplasm; Homologous Gene; Human; Inherited; Knockout Mice; Laboratories; Lead; Malignant Neoplasms; Maps; Mediating; Menin; Modeling; Modification; Molecular; Multiple Endocrine Neoplasia Type 1; Mus; Muscle hypotonia; Myeloid-Lymphoid Leukemia Protein; Null Lymphocytes; Numbers; Obesity; Organogenesis; Parathyroid gland; Pathway interactions; Patients; Phenotype; Phenylalanine-Specific tRNA; Phosphotransferases; Plasminogen Activator Inhibitor 1; Play; Process; Proliferating; Protein Overexpression; Proteins; Rate; Reporting; Resistance; Resolution; Role; Skeletal system; Small Interfering RNA; Somatic Cell; Staging; Stem cells; Syndrome; TP53 gene; Techniques; Technology; Tissue Differentiation; Tissues; Transcriptional Regulation; Tretinoin; Tumor Cell Line; Yeasts; base; cardiogenesis; cell type; clinical phenotype; density; embryonic stem cell; extracellular; fibulin 2; mRNA Decay; malignant breast neoplasm; member; morphogens; novel; periostin; promoter; response; small hairpin RNA; stem; subcutaneous; tumor; tumorigenesis; versican; wybutosine

Our laboratory employs genomic technologies for use in identifying disease-causing genes. We identified the gene (MEN1) responsible for multiple endocrine neoplasia type 1 (MEN1) syndrome in 1997. MEN1 is a familial cancer syndrome characterized by tumors in multiple endocrine tissues, primarily in the parathyroid, enteropancreas, and anterior pituitary. The amino acid sequence of the MEN1-encoded protein (menin) does not contain any identifiable structural or functional motifs. Our studies using mouse MEN1 models have revealed that the heterozygous loss of Men1 results in a tumor spectrum similar to that in humans, thus explaining the role of menin in tumorigenesis. Interestingly, homozygous loss results in early embryonic lethality (E12.5) indicating the importance of menin in early development. We have found that menin resides primarily in the nucleus, and is involved in transcriptional regulation. We are trying to elucidate the functional roles of menin in tissue differentiation, organogenesis, and development by identifying the overall changes in expression associated with the loss or overexpression of menin itself. To this end, we have generated cell lines that are null for menin in two cell types: mouse embryo fibroblasts (MEFs) and mouse embryonic stem cells (ES). Menin-null MEFs showed a significant decrease in the expression of several extracellular matrix/cell adhesion protein genes including fibulin-2 (Fbln2), periostin (Postn), and versican (Cspg2, chondroitin sulfate proteoglycan). All three of these proteins, which showed reduced expression in the absence of menin, are critical for the developing heart, and thus explain the defective heart development observed in the homozygous knockout mouse embryos. In addition, these three genes are also known targets of TGF-&#946;. Consistently, we find that TGF-&#946; responsive PAI-1 promoter is less responsive to TGF-&#946; in menin-null MEFs. Expression arrays were used to catalog the genes responding to TGF-&#946; in the presence or absence of menin. Menin interacts with MLL, the mixed lineage leukemia protein, and both are components of a COMPASS-like protein complex, regulating expression of Hox genes among others. MLL-null mouse embryonic stem (ES) cells are defective hematopoietic differentiation via dysregulation of Hox genes. Thus, it appears that menin may play a role in hematopoiesis. We generated three menin-null ES clones, and found that they were deficient in their ability to generate mature hematopoietic colonies. Formation of embryoid bodies (EB), an initial step in ES cell differentiation, was normal in menin-null cells. However, their subsequent differentiation into mature colonies was affected by the absence of menin. Re-expression of either Men1 or Hoxa9 could rescue the defect. Global expression changes in menin-null ES cells during different stages of hematopoietic differentiation are being studied to evaluate the molecular changes mediated by menin in this differentiation process leading to mesodermal lineages. We also evaluated whether loss of menin affects differentiation into endodermal lineages, using the pluripotent mouse P19 embryonic stem cells, in which differentiation processes are very well studied. We find that menin expression is induced by a natural morphogen, the retinoic acid (RA), which, at 10 nM concentration, induces endodermal differentiation in P19 cells. Interestingly, cells with shRNA-mediated reduced menin expression were resistant to endodermal differentiation when stimulated with RA. These cells proliferated at a higher rate and retained the stem cell-specific antigen SSEA-1 as in the untreated cells. On the other hand, menin overexpressing cells displayed the characteristic of an endodermal phenotype by the acquisition of cytokeratin Endo A expression, a marker for the primitive endoderm, with a concomitant loss of SSEA-1. Menins ability to induce endodermal differentiation in the absence of RA implied that menin could substitute RA by inducing a set of target genes that are RA-responsive. We have used high-density SNP arrays to molecularly define the precise deletion boundaries in patients with 2q37 deletion syndrome, characterized by several distinct physical features, including short stature, round faces, subcutaneous ossifications, brachydactyly, and other skeletal anomalies. Developmental delay, obesity, hypotonia, and autism have been reported in some cases. We found the smallest deletion (<2 Mb) in a patient with minimal clinical characteristics. We have observed three cases (out of ten) where the 2q terminal deletion is also associated with duplication of the adjacent region. A detailed evaluation of the deletion intervals for distinct clinical phenotype is being pursued. Using high-resolution BAC arrays for Array CGH, we had identified a novel gene, TRMT12, amplified and overexpressed in breast cancer cell lines and tumors. TRMT12 is a human homolog of a yeast gene encoding an enzyme that catalyzes a step in the posttranscriptional modification of a G to a highly modified base, yW (wybutosine), present in tRNAPhe. We find that human TRMT12 can substitute for the homologous yeast gene in the yW biosynthetic pathway in yeast. However, yW modification was unaffected in tRNAPhe from mouse mammary tumor model overexpressing TRMT12. Our efforts to identify the consequence of TRMT12 over expression lead to the discovery that TRMT12 regulates the expression of SMG-1, a new member of the well-known PIKK kinase family. We have demonstrated that siRNA-mediated reduced expression of TRMT12 results in increased SMG-1 expression. SMG-1 regulates non-sense mediated mRNA decay (NMD) as well as a key player in p53 mediated genotoxic stress. The functional role(s) of TRMT12 in NMD and cellular response to DNA damage is being pursued.

我们的实验室采用基因组技术来鉴定致病基因。我们于1997年确定了多发性内分泌肿瘤1型综合征的基因（MEN1）。MEN1是一种以多发内分泌组织肿瘤为特征的家族性肿瘤综合征，主要发生在甲状旁腺、肠胰腺和垂体前叶。men1编码蛋白（menin）的氨基酸序列不包含任何可识别的结构或功能基序。我们使用小鼠MEN1模型的研究表明，MEN1的杂合缺失导致与人类相似的肿瘤谱，从而解释了menin在肿瘤发生中的作用。有趣的是，纯合子缺失会导致早期胚胎致死（E12.5），这表明menin在早期发育中的重要性。我们发现menin主要存在于细胞核中，并参与转录调控。我们正试图阐明menin在组织分化、器官发生和发育中的功能作用，通过确定menin本身缺失或过度表达相关的表达的整体变化。为此，我们在小鼠胚胎成纤维细胞（mef）和小鼠胚胎干细胞（ES）两种细胞类型中产生了不含menin的细胞系。

项目成果

Pancreatic insulinomas in multiple endocrine neoplasia, type I knockout mice can develop in the absence of chromosome instability or microsatellite instability.

多发性内分泌肿瘤中的胰腺胰岛素瘤，I型基因敲除小鼠可以在没有染色体不稳定或微卫星不稳定的情况下发生。

• DOI: 10.1158/0008-5472.can-04-1648
• 发表时间: 2004
• 期刊: Cancer research
• 影响因子: 11.2
• 作者: Scacheri,PeterC;Kennedy,AlyssaL;Chin,Koei;Miller,MeghanT;Hodgson,JGraeme;Gray,JoeW;Marx,StephenJ;Spiegel,AllenM;Collins,FrancisS
• 通讯作者: Collins,FrancisS

A 700-kb physical and transcription map of the cervical cancer tumor suppressor gene locus on chromosome 11q13.

• DOI: 10.1016/j.ygeno.2005.02.014
• 发表时间: 2005-06
• 期刊: Genomics
• 影响因子: 4.4
• 作者: K. Zainabadi;Payam Benyamini;R. Chakrabarti;M. Veena;S. Chandrasekharappa;R. Gatti;E. Srivatsan

Hyperparathyroidism in hereditary syndromes: special expressions and special managements.

遗传综合征中的甲状旁腺功能亢进症：特殊表现和特殊治疗。

• 发表时间: 2002
• 期刊: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research
• 作者: Marx,StephenJ;Simonds,WilliamF;Agarwal,SunitaK;Burns,ALee;Weinstein,LeeS;Cochran,Craig;Skarulis,MonicaC;Spiegel,AllenM;Libutti,StevenK;AlexanderJr,HRichard;Chen,ClaraC;Chang,Richard;Chandrasekharappa,SettaraC;Collins,
• 通讯作者: Collins,

Characterization of frequently deleted 6q locus in prostate cancer.

前列腺癌中频繁缺失的 6q 位点的表征。

• DOI: 10.1089/dna.2006.25.597
• 发表时间: 2006
• 期刊: DNA and cell biology
• 影响因子: 3.1
• 作者: Sun,Mei;Srikantan,Vasantha;Ma,Lanfeng;Li,Jia;Zhang,Wei;Petrovics,Gyorgy;Makarem,Mazen;Strovel,JeffreyW;Horrigan,StephenG;Augustus,Meena;Sesterhenn,IsabellA;Moul,JuddW;Chandrasekharappa,Settara;Zou,Zhiqiang;Srivastava,Shiv
• 通讯作者: Srivastava,Shiv

Parathyroid gland-specific deletion of the mouse Men1 gene results in parathyroid neoplasia and hypercalcemic hyperparathyroidism.

• 发表时间: 2003-11
• 期刊: Cancer research
• 影响因子: 11.2
• 作者: S. Libutti;J. Crabtree;D. Lorang;A. Burns;C. Mazzanti;S. Hewitt;Sarah O'Connor;J. Ward;M. Emmert-Buck;A. Remaley;Marshall S. Miller;E. Turner;H. Alexander;A. Arnold;S. Marx;F. Collins;A. Spiegel