公开(公告)号：EP1572867B1

公开(公告)日：2014-07-23

申请号：EP02728495.9

申请日：2002-04-10

申请人： The Johns Hopkins University

发明人： CARSON-WALTER, Eleanor ,  ST. CROIX, Brad ,  KINZLER, Kenneth, W. ,  VOGELSTEIN, Bert

IPC分类号： C07K14/705 ,  G01N33/53 ,  C07K16/30

CPC分类号： C07K14/705 ,  A61K2039/505 ,  C07K14/4748 ,  C07K16/28 ,  G01N33/5023 ,  G01N33/68 ,  G01N2333/4703 ,  G01N2500/04 ,  G01N2800/7014 ,  G01N2800/7028

摘要： To gain a better understanding of tumor angiogenesis, new techniques for isolating endothelial cells (ECs) and evaluating gene expression patterns were developed. When transcripts from ECs derived from normal and malignant colorectal tissues were compared with transcripts from non-endothelial cells, over 170 genes predominantly expressed in the endothelium were identified. Comparison between normal- and tumor-derived endothelium revealed 79 differentially expressed genes, including 46 that were specifically elevated in tumor-associated endothelium. Experiments with representative genes from this group demonstrated that most were similarly expressed in the endothelium of primary lung, breast, brain, and pancreatic cancers as well as in metastatic lesions of the liver. These results demonstrate that neoplastic and normal endothelium in humans are distinct at the molecular level, and have significant implications for the development of anti-angiogenic therapies in the future.

公开(公告)号：EP2723896A2

公开(公告)日：2014-04-30

申请号：EP12802288.6

申请日：2012-06-22

申请人： The Johns Hopkins University

发明人： VOGELSTEIN, Bert ,  KINZLER, Kenneth W. ,  WU, Jian ,  DIAZ, Luis ,  PAPADOPOULOS, Nickolas ,  MATTHAEI, Hanno ,  HRUBAN, Ralph ,  MAITRA, Anirban

IPC分类号： C12Q1/68 ,  C12N15/11

CPC分类号： C12Q1/6886 ,  C12Q2600/112 ,  C12Q2600/156

摘要： To help reveal the pathogenesis of these lesions, we purified the DNA from Intraductal Papillary Mucinous Neoplasm (IPMN) cyst fluids from 19 patients and searched for mutations in 169 genes commonly altered in human cancers. We identified recurrent mutations at codon 201 of GNAS. We found that GNAS mutations were present in 66% of IPMNs and that either KRAS or GNAS mutations could be identified in 96%. In eight cases, we could investigate invasive adenocarcinomas that developed in association with IPMNs containing GNAS mutations. In seven of these eight cases, the GNAS mutations present in the IPMNs were also found in the invasive lesion. GNAS mutations were not found in other types of cystic neoplasms of the pancreas or in invasive adenocarcinomas not associated with IPMNs. These data suggest that GNAS mutations can inform the diagnosis and management of patients with cystic pancreatic lesions.

摘要翻译： 为了揭示这些病变的发病机制，我们从19名患者的导管内乳头状粘液性肿瘤（IPMN）囊肿液中纯化了DNA，并在人类癌症中通常改变的169种基因中进行了突变检测。 我们在GNAS的201号密码子处鉴定了复发性突变。 我们发现GNAS突变存在于66％的IPMNs中，KRAS或GNAS突变可以在96％中鉴定。 在8例中，我们可以研究与含有GNAS突变的IPMN相关的侵袭性腺癌。 在这8例中有7例中，存在于IPMNs中的GNAS突变也在侵袭性损伤中发现。 在其他类型的胰腺囊性肿瘤或与IPMN无关的侵袭性腺癌中未发现GNAS突变。 这些数据表明，GNAS突变可以为囊性胰腺病变患者的诊断和治疗提供依据。

公开(公告)号：EP2536854A2

公开(公告)日：2012-12-26

申请号：EP11745196.3

申请日：2011-02-17

申请人： The Johns Hopkins University

发明人： VOGELSTEIN, Bert ,  KINZLER, Kenneth W. ,  VELCULESCU, Victor ,  DIAZ, Luis ,  LEARY, Rebecca J.

IPC分类号： C12Q1/68 ,  C12N15/11 ,  C40B40/06

CPC分类号： C12Q1/6886 ,  C12Q2600/156

摘要： Clinical management of human cancer is dependent on the accurate monitoring of residual and recurrent tumors. We have developed a method, called personalized analysis of rearranged ends (PARE), which can identify translocations in solid tumors. Analysis of four colorectal and two breast cancers revealed an average of nine rearranged sequences (range 4 to 15) per tumor. Polymerase chain reaction with primers spanning the breakpoints were able to detect mutant DNA molecules present at levels lower than 0.001% and readily identified mutated circulating DNA in patient plasma samples. This approach provides an exquisitely sensitive and broadly applicable approach for the development of personalized biomarkers to enhance the clinical management of cancer patients.

公开(公告)号：EP1948816B1

公开(公告)日：2011-12-07

申请号：EP06817236.0

申请日：2006-10-20

申请人： The Johns Hopkins University

发明人： VOGELSTEIN, Bert ,  DIEHL, Frank ,  KINZLER, Kenneth, W. ,  LI, Ming

IPC分类号： C12P19/34

CPC分类号： C12Q1/6858 ,  G01R31/025 ,  G01R31/3277 ,  C12Q2565/537 ,  C12Q2563/155 ,  C12Q2527/125

摘要： Improvements on the basic method used for BEAMing increase sensitivity and increase the signal-to-noise ratio. The improvements have permitted the determination of intrinsic error rates of various DNA polymerases and have permitted the detection of rare and subtle mutations in DNA isolated from plasma of cancer patients.

摘要翻译： 用于BEAMing的基本方法的改进提高了灵敏度并提高了信噪比。 这些改进允许确定各种DNA聚合酶的固有错误率，并允许检测从癌症患者血浆中分离的DNA中罕见且微妙的突变。

公开(公告)号：EP1430071B1

公开(公告)日：2011-04-06

申请号：EP02773302.1

申请日：2002-09-09

申请人： THE JOHNS HOPKINS UNIVERSITY SCHOOL OF MEDICINE

发明人： BUCKHAULTS, Phillip ,  KINZLER, Kenneth, W. ,  VOGELSTEIN, Bert

IPC分类号： C07H21/04 ,  C12N9/48 ,  C07K14/495

CPC分类号： C07K14/495 ,  C07K14/47 ,  C07K14/705

摘要： Serial analysis of gene expression (SAGE) was used to identify transcripts encoding secreted or cell-surface proteins that were expressed in benign and malignant tumors of the colorectum. A total of 290,394 tags were analyzed from normal, adenomatous and cancerous colonic epithelium. Of the 21,343 different transcripts observed, 957 were found to be differentially expressed between normal and adenoma or between normal and cancer. Forty-nine transcripts were elevated ≥ 20-fold in adenomas, 40 transcripts were elevated ≥ 20-fold in cancers, and nine transcripts were elevated ≥ 20-fold in both. Product of six these nine transcripts (TGFBI, LYS, RDP, MIC-1, REGA, and DEHL) were predicted to be secreted or to reside on the cell surface and these were analyzed in more detail. The abnormal expression levels predicted by SAGE were confirmed by quantitative PCR analyses of each of these six genes. Moreover, the cell types responsible for the elevated expression were identified by in situ hybridization and by PCR analyses of epithelial cells immunoaffinity purified from primary tumors.

公开(公告)号：EP0972037B1

公开(公告)日：2006-08-16

申请号：EP98912994.5

申请日：1998-03-20

申请人： THE JOHNS HOPKINS UNIVERSITY ,  Kylix B.V.

发明人： BARKER, Nick ,  CLEVERS, Hans ,  KINZLER, Kenneth, W. ,  KORINEK, Vladimir ,  MORIN, Patrice, J. ,  SPARKS, Andrew, B. ,  VOGELSTEIN, Bert

IPC分类号： C12N15/12 ,  C07K14/47 ,  C12Q1/68 ,  G01N33/50 ,  A61K38/17 ,  A61K48/00

CPC分类号： C07K14/47 ,  A61K38/00 ,  A61K48/00 ,  C07K14/4705 ,  C12Q1/6897

摘要： The APC tumor suppressor protein binds to beta -catenin, a protein recently shown to interact with Tcf/Lef transcription factors. Here, the gene encoding a Tcf family member that is expressed in colonic epithelium (hTcf-4) was cloned and characterized. hTcf-4 transactivates transcription only when associated with beta -catenin. Nuclei of APC colon carcinoma cells were found to contain a stable beta -catenin-hTCF-4 complex that was constitutively active, as measured by transcription of a Tcf reporter gene. Reintroduction of APC removed beta -catenin from hTcf4 and abrogated the transcriptional transactivation. Constitutive transcription of TCF target genes, caused by loss of APC function, may be a crucial event in the early transformation of colonic epithelium. It is also shown here that the products of mutant APC genes found in colorectal tumors are defective in regulating beta -catenin/Tcf-4 transcriptional activation. Furthermore, colorectal tumors with intact APC genes were shown to contain subtle activating mutations of beta -catenin that altered functionally significant phosphorylation sites. These results indicate that regulation of beta -catenin is critical to APC's tumor suppressive effect and that this regulation can be circumvented by mutations in either APC or beta -catenin.

公开(公告)号：EP1255856B1

公开(公告)日：2005-03-09

申请号：EP00952304.4

申请日：2000-07-31

申请人： THE JOHNS HOPKINS UNIVERSITY

发明人： KINZLER, Kenneth, W. ,  VOGELSTEIN, Bert

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6886 ,  C12Q1/6818 ,  C12Q1/6851 ,  C12Q1/686 ,  C12Q1/6874 ,  C12Q2600/156 ,  C12Q2600/158

摘要： The identification of pre-defined mutations expected to be present in a minor fraction of a cell population is important for a variety of basic research and clinical applications. The exponential, analog nature of the polymerase chain reaction is transformed into a linear, digital signal suitable for this purpose. Single molecules can be isolated by dilution and individually amplified; each product is then separately analyzed for the presence of mutations. The process provides a reliable and quantitative measure of the proportion of variant sequences within a DNA sample.

公开(公告)号：EP1104475B1

公开(公告)日：2004-05-26

申请号：EP99943741.1

申请日：1999-08-20

申请人： The Johns Hopkins University School of Medicine

发明人： HE, Tong-Chuan ,  VOGELSTEIN, Bert ,  KINZLER, Kenneth, W.

IPC分类号： C12N15/63 ,  C12Q1/68 ,  C07K14/27

CPC分类号： C07K14/47 ,  A61K38/00 ,  A61K48/00 ,  C07K14/4705 ,  C12Q1/6897

摘要： The APC tumor suppressor protein binds to β-catenin, a protein recently shown to interact with Tcf/Lef transcription factors. Here, the gene encoding a Tcf family member that is expressed in colonic epithelium (hTcf-4) was cloned and characterized. hTcf-4 transactivates transcription only when associated with β-catenin. Nuclei of APC-/- colon carcinoma cells were found to contain a stable β-catenin-hTCF-4 complex that was constitutively active, as measured by transcription of a Tcf reporter gene. Reintroduction of APC removed β-catenin from hTcf4 and abrogated the transcriptional transactivation. Constitutive transcription of TCF target genes, caused by loss of APC function, may be a crucial event in the early transformation of colonic epithelium. It is also shown here that the products of mutant APC genes found in colorectal tumors are defective in regulating β-catenin/Tcf-4 transcriptional activation. Furthermore, colorectal tumors with intact APC genes were shown to contain subtle activating mutations of β-catenin that altered functionally significant phosphorylation sites. These results indicate that regulation of β-catenin is critical to APC"s tumor suppressive effect and that this regulation can be circumvented by mutations in either APC or β-catenin.

公开(公告)号：EP1104492B1

公开(公告)日：2004-02-11

申请号：EP99943742.9

申请日：1999-08-20

申请人： THE JOHNS HOPKINS UNIVERSITY SCHOOL OF MEDICINE

发明人： VOGELSTEIN, Bert ,  KINZLER, Kenneth, W. ,  POLYAK, Kornelia

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6809 ,  C12Q1/6886 ,  C12Q2600/156

摘要： The accumulation of homoplasmic somatic mutations has been observed in the mitochondrial DNA of certain tumor cells. The presence or a recurrence of a tumor can be detected by determining the presence of single basepair mutations in the mitochondrial genome from a cell sample of a patient.

公开(公告)号：EP0847450B1

公开(公告)日：2003-11-05

申请号：EP96930551.5

申请日：1996-08-23

申请人： THE JOHNS HOPKINS UNIVERSITY

发明人： VOGELSTEIN, Bert ,  KINZLER, Kenneth, W. ,  PAPADOPOULOS, Nickolas

IPC分类号： C12Q1/68 ,  C12P21/00 ,  G01N33/50 ,  G01N33/574 ,  G01N33/68

CPC分类号： G01N33/68 ,  C12Q1/6827 ,  G01N33/5005 ,  C12Q2561/107

摘要： A diagnostic strategy for detection of inherited diseases caused by germline mutations is based on somatic cell hybridization. Each allele of a human gene involved in the inherited disease is isolated in a somatic cell hybrid. The products of the isolated human allele are then observed in the absence of the other allele of the human.