公开(公告)号：US09976184B2

公开(公告)日：2018-05-22

申请号：US14128478

申请日：2012-06-22

申请人： Bert Vogelstein ,  Kenneth W. Kinzler ,  Jian Wu ,  Luis Diaz ,  Nickolas Papadopoulos ,  Hanno Matthaei ,  Ralph Hruban ,  Anirban Maitra

发明人： Bert Vogelstein ,  Kenneth W. Kinzler ,  Jian Wu ,  Luis Diaz ,  Nickolas Papadopoulos ,  Hanno Matthaei ,  Ralph Hruban ,  Anirban Maitra

IPC分类号： C12Q1/68 ,  C12P19/34

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.

公开(公告)号：US20140179538A1

公开(公告)日：2014-06-26

申请号：US14128478

申请日：2012-06-22

申请人： Bert Vogelstein ,  Kenneth W. Kinzler ,  Jian Wu ,  Luis Diaz ,  Nickolas Papadopoulos ,  Hanno Matthaei ,  Ralph Hruban ,  Anirban Maitra

发明人： Bert Vogelstein ,  Kenneth W. Kinzler ,  Jian Wu ,  Luis Diaz ,  Nickolas Papadopoulos ,  Hanno Matthaei ,  Ralph Hruban ,  Anirban Maitra

IPC分类号： C12Q1/68

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DNA，并搜索了在人类癌症中通常改变的169种基因中的突变。 我们发现GNAS密码子201的复发性突变。 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，我们可以调查与含有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突变可以通过诊断和管理囊性胰腺病变患者。

公开(公告)号：US09982306B2

公开(公告)日：2018-05-29

申请号：US14129850

申请日：2012-06-28

申请人： Hai Yan ,  Darell Bigner ,  Bert Vogelstein ,  Kenneth W. Kinzler ,  Alan Meeker ,  Ralph Hruban ,  Nickolas Papadopoulos ,  Luis Diaz ,  Yuchen Jiao

发明人： Hai Yan ,  Darell Bigner ,  Bert Vogelstein ,  Kenneth W. Kinzler ,  Alan Meeker ,  Ralph Hruban ,  Nickolas Papadopoulos ,  Luis Diaz ,  Yuchen Jiao

IPC分类号： G01N33/68 ,  C12Q1/68 ,  G01N33/574 ,  C07K16/40 ,  C12N15/113

CPC分类号： C12Q1/6886 ,  C07K16/40 ,  C12N15/1137 ,  C12Q2600/118 ,  C12Q2600/156 ,  G01N33/57407

摘要： We determined the sequence of ATRX and DAXX in 447 cancers from various sites. We found mutations most commonly in pediatric glioblastoma multiformae (GBM) (11.1%), adult GBM (6.5%), oligodendrogliomas (7.7%) and medulloblastomas (1.5%); and showed that Alternative Lengthening of Telomeres (ALT), a telomerase-independent telomere maintenance mechanism found in cancers that have not activated telomerase, perfectly correlated with somatic mutations of either gene. In contrast, neuroblastomas, and adenocarcinomas of the ovary, breast, and pancreas were negative for mutations in ATRX and DAXX. Alterations in ATRX or DAXX define a specific molecular pathway that is closely associated with an alternative telomere maintenance function in human cancers.

公开(公告)号：US20140227271A1

公开(公告)日：2014-08-14

申请号：US14129850

申请日：2012-06-28

申请人： Hai Yan ,  Darell Bigner ,  Bert Vogelstein ,  Kenneth W. Kinzler ,  Alan Meeker ,  Ralph Hruban ,  Nickolas Papadopoulos ,  Luis Diaz ,  Yuchen Jiao

发明人： Hai Yan ,  Darell Bigner ,  Bert Vogelstein ,  Kenneth W. Kinzler ,  Alan Meeker ,  Ralph Hruban ,  Nickolas Papadopoulos ,  Luis Diaz ,  Yuchen Jiao

IPC分类号： C12Q1/68 ,  C07K16/40 ,  C12N15/113 ,  G01N33/574

CPC分类号： C12Q1/6886 ,  C07K16/40 ,  C12N15/1137 ,  C12Q2600/118 ,  C12Q2600/156 ,  G01N33/57407

摘要： We determined the sequence of ATRX and DAXX in 447 cancers from various sites. We found mutations most commonly in pediatric glioblastoma multiformae (GBM) (11.1%), adult GBM (6.5%), oligodendrogliomas (7.7%) and medulloblastomas (1.5%); and showed that Alternative Lengthening of Telomeres (ALT), a telomerase-independent telomere maintenance mechanism found in cancers that have not activated telomerase, perfectly correlated with somatic mutations of either gene. In contrast, neuroblastomas, and adenocarcinomas of the ovary, breast, and pancreas were negative for mutations in ATRX and DAXX. Alterations in ATRX or DAXX define a specific molecular pathway that is closely associated with an alternative telomere maintenance function in human cancers.

摘要翻译： 我们确定了来自各个位点的447例癌症中ATRX和DAXX的序列。 我们发现多形性小儿多形性成胶质细胞瘤（GBM）（11.1％），成人GBM（6.5％），少突胶质细胞瘤（7.7％）和成神经管细胞瘤（1.5％）中最常见的突变; 并且表明，在未激活端粒酶的癌症中发现端粒酶不依赖端粒维持机制的替代延长端粒（ALT）与任一基因的体细胞突变完全相关。 相比之下，成骨细胞瘤和卵巢，乳腺和胰腺腺癌对于ATRX和DAXX中的突变是阴性的。 ATRX或DAXX中的改变定义了与人类癌症中替代端粒维持功能密切相关的特定分子途径。

公开(公告)号：US20120164638A1

公开(公告)日：2012-06-28

申请号：US13263020

申请日：2010-04-06

申请人： Bert Vogelstein ,  Kenneth W. Kinzler ,  Meng Li ,  Luis Diaz ,  Nickolas Papadopoulos ,  Sanford Markowitz

发明人： Bert Vogelstein ,  Kenneth W. Kinzler ,  Meng Li ,  Luis Diaz ,  Nickolas Papadopoulos ,  Sanford Markowitz

IPC分类号： C12Q1/68 ,  C07H21/04

CPC分类号： C12Q1/6816 ,  C12Q1/6886 ,  C12Q2600/154 ,  C12Q2565/501 ,  C12Q2527/125 ,  C12Q2523/125

摘要： Abnormal DNA methylation can be used as a biomarker in cancer patients. For such purposes, it is important to determine precisely the fraction of methylated molecules in an analyzed sample. A technology we term Methyl-BEAMing achieves this goal. Individual bisulfite-treated DNA molecules can be PCR-amplified within aqueous nanocompartments containing beads, resulting in a population of beads each containing thousands of copies of the template molecule. After hybridization with probes specific for methylated sequences, the beads can be analyzed by flow cytometry. This approach enables detection and enumeration of one methylated molecule in a population of ˜5000 unmethylated molecules. Methyl-BEAMing provides digital quantification of rare methylation events and is generally applicable to the assessment of methylated genes in clinical samples.

摘要翻译： DNA甲基化异常可用作癌症患者的生物标志物。 为此目的，重要的是精确测定分析样品中甲基化分子的分数。 我们称之为甲基BEAMing的技术实现了这一目标。 单独的亚硫酸氢盐处理的DNA分子可以在含有珠粒的含水纳米间隔内进行PCR扩增，导致每个珠粒群含有数千份拷贝的模板分子。 与特异于甲基化序列的探针杂交后，可以通过流式细胞术分析珠粒。 这种方法能够检测和计数一个约5000个非甲基化分子的群体中的一个甲基化分子。 甲基BEAMing提供罕见甲基化事件的数字量化，通常适用于临床样品中甲基化基因的评估。

公开(公告)号：US10144971B2

公开(公告)日：2018-12-04

申请号：US13977810

申请日：2012-01-04

申请人： Bert Vogelstein ,  Kenneth W Kinzler ,  Victor Velculescu ,  Luis Diaz ,  Nikolas Papadopoulos ,  Yuchen Jiao ,  Ralph Hruban

发明人： Bert Vogelstein ,  Kenneth W Kinzler ,  Victor Velculescu ,  Luis Diaz ,  Nikolas Papadopoulos ,  Yuchen Jiao ,  Ralph Hruban

IPC分类号： C12Q1/68 ,  C12P19/34 ,  C12Q1/6886 ,  G01N33/574

摘要： Pancreatic Neuroendocrine Tumors (PanNETs) are a rare but clinically important form of pancreatic neoplasia. To explore the genetic basis of PanNETs, we determined the exomic sequences of ten non-familial PanNETs and then screened the most commonly mutated genes in 58 additional PanNETs. Remarkably, the most frequently mutated genes specify proteins implicated in chromatin remodeling: 44% of the tumors had somatic inactivating mutations in MEN-1, which encodes menin, a component of a histone methyltransferase complex; and 43% had mutations in genes encoding either of the two subunits of a transcription/chromatin remodeling complex consisting of DAXX (death-domain associated protein) and ATRX (alpha thalassemia/mental retardation syndrome X-linked). Clinically, mutations in the MEN1 and DAXX/ATRX genes were associated with better prognosis. We also found mutations in genes in the mTOR (mammalian target of rapamycin) pathway in 14% of the tumors, a finding that could potentially be used to stratify patients for treatment with mTOR inhibitors.

公开(公告)号：US20140045881A1

公开(公告)日：2014-02-13

申请号：US13977810

申请日：2012-01-04

申请人： Bert Vogelstein ,  Kenneth W. Kinzler ,  Victor Velculescu ,  Luis Diaz ,  Nikolas Papadopoulos ,  Yuchen Jiao ,  Ralph Hruban

发明人： Bert Vogelstein ,  Kenneth W. Kinzler ,  Victor Velculescu ,  Luis Diaz ,  Nikolas Papadopoulos ,  Yuchen Jiao ,  Ralph Hruban

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6886 ,  C12Q2600/106 ,  C12Q2600/112 ,  C12Q2600/118 ,  C12Q2600/156 ,  G01N33/57438 ,  G01N2800/52

摘要： Pancreatic Neuroendocrine Tumors (PanNETs) are a rare but clinically important form of pancreatic neoplasia. To explore the genetic basis of PanNETs, we determined the exomic sequences of ten non-familial PanNETs and then screened the most commonly mutated genes in 58 additional PanNETs. Remarkably, the most frequently mutated genes specify proteins implicated in chromatin remodeling: 44% of the tumors had somatic inactivating mutations in MEN-1, which encodes menin, a component of a histone methyltransferase complex; and 43% had mutations in genes encoding either of the two subunits of a transcription/chromatin remodeling complex consisting of DAXX (death-domain associated protein) and ATRX (alpha thalassemia/mental retardation syndrome X-linked). Clinically, mutations in the MEN1 and DAXX/ATRX genes were associated with better prognosis. We also found mutations in genes in the mTOR (mammalian target of rapamycin) pathway in 14% of the tumors, a finding that could potentially be used to stratify patients for treatment with mTOR inhibitors.

摘要翻译： 胰腺神经内分泌肿瘤（Pannets）是一种罕见但临床上重要的胰腺肿瘤形式。 为了探索PanNETs的遗传基础，我们确定了十个非家族PanNETs的外显子序列，然后筛选了58个额外的PanNET中最常见的突变基因。 值得注意的是，最常突变的基因指定涉及染色质重塑的蛋白质：44％的肿瘤在MEN-1中具有体细胞失活突变，其编码menin，组蛋白甲基转移酶复合物的组分; 43％在编码由DAXX（死亡相关蛋白）和ATRX（α地中海贫血/智力迟钝综合征X连锁）组成的转录/染色质重塑复合物的两个亚基之一的基因中具有突变。 在临床上，MEN1和DAXX / ATRX基因的突变与更好的预后相关。 我们还发现在14％的肿瘤中mTOR（雷帕霉素的哺乳动物靶标）途径的基因突变，这种发现可能用于分层患者用mTOR抑制剂治疗。

公开(公告)号：US20100041048A1

公开(公告)日：2010-02-18

申请号：US12512585

申请日：2009-07-30

申请人： Frank DIEHL ,  Luis Diaz ,  Kenneth W. Kinzler ,  Bert Vogelstein ,  Kerstin Schmidt

发明人： Frank DIEHL ,  Luis Diaz ,  Kenneth W. Kinzler ,  Bert Vogelstein ,  Kerstin Schmidt

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6886 ,  C12Q2525/197 ,  C12Q2535/131 ,  C12Q2549/119 ,  C12Q2600/112 ,  C12Q2600/118 ,  C12Q2600/136

摘要： DNA containing somatic mutations is highly tumor specific and thus, in theory, can provide optimum markers. However, the number of circulating mutant gene fragments is small compared to the number of normal circulating DNA fragments, making it difficult to detect and quantify them with the sensitivity required for meaningful clinical use. We apply a highly sensitive approach to quantify circulating tumor DNA (ctDNA) in body samples of patients. Measurements of ctDNA can be used to reliably monitor tumor dynamics in subjects with cancer, especially those who are undergoing surgery or chemotherapy. This personalized genetic approach can be generally applied.

摘要翻译： 含有体细胞突变的DNA具有高度的肿瘤特异性，因此在理论上可以提供最佳的标记。 然而，循环突变基因片段的数量与正常循环DNA片段的数量相比较小，使得难以用有意义的临床使用所需的灵敏度来检测和量化它们。 我们应用高度敏感的方法来量化患者身体样本中的循环肿瘤DNA（ctDNA）。 ctDNA的测量可用于可靠地监测癌症患者的肿瘤动态，特别是正在接受手术或化疗的患者。 这种个性化的遗传方法一般可以应用。

公开(公告)号：US20130210645A1

公开(公告)日：2013-08-15

申请号：US13579964

申请日：2011-02-17

申请人： Bert Volgelstein ,  Kenneth W. Kinzler ,  Victor Velculescu ,  Luis Diaz ,  Rebecca J. Leary

发明人： Bert Volgelstein ,  Kenneth W. Kinzler ,  Victor Velculescu ,  Luis Diaz ,  Rebecca J. Leary

IPC分类号： C12Q1/68

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.

摘要翻译： 人类癌症的临床管理取决于对残留和复发性肿瘤的准确监测。 我们开发了一种称为重排末端（PARE）的个性化分析方法，可以识别实体瘤易位。 对4例结直肠癌和2例乳腺癌的分析显示，每个肿瘤平均有9个重排序列（范围4至15）。 与跨越断点的引物的聚合酶链反应能够检测出低于0.001％的突变型DNA分子，并且容易鉴定出患者血浆样品中突变的循环DNA。 这种方法为个性化生物标志物的开发提供了一种非常灵敏和广泛适用的方法，以加强癌症患者的临床管理。

公开(公告)号：US20130323167A1

公开(公告)日：2013-12-05

申请号：US13880608

申请日：2011-10-20

申请人： Bert Vogelstein ,  Yuan Qiao ,  Xin Huang ,  Kenneth Kinzler ,  Shibin Zhou ,  Luis Diaz

发明人： Bert Vogelstein ,  Yuan Qiao ,  Xin Huang ,  Kenneth Kinzler ,  Shibin Zhou ,  Luis Diaz

IPC分类号： A61K51/10

CPC分类号： A61K51/1045 ,  A61K9/1271 ,  A61K39/395 ,  A61K45/06 ,  A61K51/1009 ,  C07K16/2887 ,  C07K16/40 ,  A61K2300/00

摘要： Several agents capable of inducing vascular responses akin to those observed in inflammatory processes enhance the accumulation of nanoparticles in tumors. Exemplary vascular-active agents include a bacterium, a pro-inflammatory cytokine, and microtubule-destabilizing drugs. Such agents can increase the tumor to blood ratio of radioactivity by more than 20-fold compared to nanoparticles alone. Moreover, vascular-active agents dramatically improved the therapeutic effect of nanoparticles containing radioactive isotopes or chemotherapeutic agents.

摘要翻译： 能够诱导与炎症过程中观察到的血管反应类似的几种药物增强纳米颗粒在肿瘤中的积累。 示例性的血管活性剂包括细菌，促炎细胞因子和微管不稳定药物。 与单独的纳米颗粒相比，这样的药物可以将放射性的肿瘤与血液的比例增加20倍以上。 此外，血管活性剂显着改善了含有放射性同位素或化学治疗剂的纳米颗粒的治疗效果。