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1. 发明申请

WO0065077A8 TRIPLE HYBRID AMPLICON VECTOR SYSTEMS TO GENERATE RETROVIRAL PACKAGING LINES 审中-公开
标题翻译：三重混合放大器矢量系统生成回流包装线
公开(公告)号：WO0065077A8
公开(公告)日：2001-10-11
申请号：PCT/US0010669
申请日：2000-04-21
申请人： GEN HOSPITAL CORP , SENA ESTEVES MIGUEL , BREAKEFIELD XANDRA O , SAEKI YOSHINAGA
发明人： SENA-ESTEVES MIGUEL , BREAKEFIELD XANDRA O , SAEKI YOSHINAGA
IPC分类号： A61K48/00 , C07K14/15 , C12N5/10 , C12N15/38 , C12N15/867 , C12N15/864 , C12N15/63 , C12N15/64 , C12N15/869
CPC分类号： C12N7/00 , A61K48/00 , C07K14/005 , C12N15/86 , C12N2740/13022 , C12N2740/13043 , C12N2740/13052 , C12N2799/021
摘要： The present invention relates to a triple hybrid vector amplicon system comprising genetic elements derived from Herpes Simplex Virus (HSV), Epstein-Barr Virus (EBV) or Adeno-Associated Virus (AAV), and a retrovirus. The vector was developed to stably transform cells, both in culture or in vivo, into retrovirus packaging cells in a single step. This step can be accomplished both by transfection using liposomes, electroporation, calcium phosphate, or any other methodology used to transfer naked or complexed DNA into cells or by infection when the vector is packaged as an amplicon vector in HSV virions. In one embodiment, the system takes advantage of the host range and retention properties of HSV/EBV hybrid amplicons to efficiently convert cells to retrovirus vector producer cells after single-step transduction. Retrovirus genes gag-pol and env (GPE) and retroviral vector sequences were modified to minimize sequence overlap and cloned into an HSV/EBV hybrid amplicon. In a second embodiment, retrovirus gag-pol and env genes and a retrovirus vector carrying lacZ, were cloned into HSV/AAV hybrid amplicons. These hybrid amplicon vector systems hold great potential for the generation of new retrovirus packaging lines derived from cells that due to their migratory, tumor or tissue targeting properties, can expand the spatial distribution of gene delivery by retrovirus vectors in vivo.
摘要翻译：本发明涉及包含衍生自单纯疱疹病毒（HSV），爱泼斯坦 - 巴尔病毒（EBV）或腺相关病毒（AAV））和逆转录病毒的遗传元件的三重杂交载体扩增子系统。开发该载体以在单一步骤中将培养物或体内的细胞稳定地转化成逆转录病毒包装细胞。该步骤可以通过使用脂质体，电穿孔，磷酸钙或用于将裸露或复合DNA转移到细胞中的任何其它方法或当将载体作为扩增子载体包装在HSV病毒粒子中时通过感染来实现。在一个实施方案中，该系统利用HSV / EBV杂交扩增子的宿主范围和保留性质，以在单步转导后有效地将细胞转化为逆转录病毒载体生产细胞。逆转录病毒基因gag-pol和env（GPE）和逆转录病毒载体序列被修饰以最小化序列重叠并克隆到HSV / EBV杂交扩增子中。在第二个实施方案中，将逆转录病毒gag-pol和env基因和携带lacZ的逆转录病毒载体克隆到HSV / AAV杂交扩增子中。这些杂交扩增子载体系统具有巨大的潜力，用于产生由细胞衍生的新的逆转录病毒包装线，由于其迁移，肿瘤或组织靶向特性，可以扩大反转录病毒载体在体内的基因递送的空间分布。

2. 发明申请

WO0034497A3 ENHANCED PACKAGING OF HERPES VIRUS AMPLICONS AND GENERATION OF RECOMBINANT VIRUS VECTORS 审中-公开
标题翻译：增强的病毒片段的包装和重组病毒载体的产生
公开(公告)号：WO0034497A3
公开(公告)日：2000-11-09
申请号：PCT/US9929120
申请日：1999-12-09
申请人： BREAKEFIELD XANDRA O , CHIOCCA E ANTONIO , SAEKI YOSHINAGA , FRAEFEL CORNEL , TOBLER KURT , ACKERMANN MATHIAS , SUTER MARK , ADEMA GOSSE J , SHORTMAN KEN
发明人： BREAKEFIELD XANDRA O , CHIOCCA E ANTONIO , SAEKI YOSHINAGA , FRAEFEL CORNEL , TOBLER KURT , ACKERMANN MATHIAS , SUTER MARK , ADEMA GOSSE J , SHORTMAN KEN
IPC分类号： A61K39/00 , A61K39/245 , A61K48/00 , C12N15/33 , C12N15/38 , C12N15/869
CPC分类号： C12N15/86 , A61K39/00 , A61K39/12 , A61K39/245 , A61K48/00 , A61K2039/5256 , A61K2039/53 , A61K2039/545 , C12N2710/16634 , C12N2710/16643 , C12N2800/204
摘要： The present invention relates to an enhanced and simplified herpes virus amplicon packaging system and method of generating recombinant herpes virus vectors by "pac rescue". In one embodiment of this packaging system, the HSW-1 genome is cloned and maintained as a single-copy, F-plasmid-based BAC in E.coli. Such a plasmid, containing the HSV-1 genome deleted for the pac signals, did not generate replication-competent progeny virus upon transfection into mammalian cells, but rather, supported the packaging of co-transfected amplicon DNA which contained a functional pac signal. In another embodiment, the HSV genome cloned in BAC has an intact pac signal, however, the helper HSV genome is made packaging defective by increasing or decreasing the size of the BAC. This method involves the co-introduction of DNA fragments containing a pac site flanked by homologous HSV sequences and pac-deleted helper virus genome to permissive cells. In another embodiment, any kind of modification can be introduced into the viral genome through homologous recombination in bacteria.
摘要翻译：本发明涉及一种增强和简化的疱疹病毒扩增子包装系统和通过“pac拯救”产生重组疱疹病毒载体的方法。在该包装系统的一个实施方案中，将HSW-1基因组克隆并维持为大肠杆菌中的基于单拷贝，F-质粒的BAC。含有针对pac信号缺失的HSV-1基因的这种质粒在转染哺乳动物细胞后没有产生复制能力的后代病毒，而是支持包含功能性pac信号的共转染的扩增子DNA的包装。在另一个实施方案中，克隆在BAC中的HSV基因组具有完整的pac信号，然而，辅助HSV基因组通过增加或减少BAC的大小而被制成包装有缺陷的。该方法涉及将含有同源HSV序列和pac缺失的辅助病毒基因组的Pac位点的DNA片段共同引入允许的细胞。在另一个实施方案中，可以通过细菌中的同源重组将任何种类的修饰引入到病毒基因组中。

3. 发明申请

WO2012031008A2 CANCER-RELATED BIOLOGICAL MATERIALS IN MICROVESICLES 审中-公开
标题翻译：与微生物相关的癌症相关生物材料
公开(公告)号：WO2012031008A2
公开(公告)日：2012-03-08
申请号：PCT/US2011050041
申请日：2011-08-31
申请人： GEN HOSPITAL CORP , SKOG JOHAN KARL OLOV , BALAJ LEONORA , NOERHOLM MIKKEL , BREAKEFIELD XANDRA O
发明人： SKOG JOHAN KARL OLOV , BALAJ LEONORA , NOERHOLM MIKKEL , BREAKEFIELD XANDRA O
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6886 , C12Q1/6806 , C12Q2600/106 , C12Q2600/112 , C12Q2600/118 , C12Q2600/154 , C12Q2600/156 , C12Q2600/158 , C12Q2600/178 , C12Q2563/161
摘要： Disclosed herein are methods for assaying a biological sample from a subject by analyzing components of microvesicle fractions in aid of risk, diagnosis, prognosis or monitoring of, or directing treatment of the subject for, a disease or other medical condition in the subject. Also disclosed are methods of treatment and identifying biomarkers using a microvesicle fraction of a subject. Kits, pharmaceutical compositions, and profiles related to the methods are also disclosed.
摘要翻译：本文公开了通过分析受试者的疾病或其他医学病症的风险，诊断，预后或对受试者的治疗来分析来自受试者的生物样品的方法。还公开了使用受试者的微泡级分的治疗和鉴定生物标志物的方法。还公开了与方法相关的试剂盒，药物组合物和特征。

4. 发明申请

WO2008066608A8 A SECRETED LUCIFERASE FLUORESCENT PROTEIN CONJUGATE NUCLEIC ACID CONSTRUCT AND USES THEREOF 审中-公开
标题翻译：一种秘密的LUCIFERASE荧光蛋白缀合物核酸构建及其应用
公开(公告)号：WO2008066608A8
公开(公告)日：2008-12-24
申请号：PCT/US2007021810
申请日：2007-10-12
申请人： GEN HOSPITAL CORP , TANNOUS BAKHOS A , BREAKEFIELD XANDRA O , HEWETT JEFFREY W
发明人： TANNOUS BAKHOS A , BREAKEFIELD XANDRA O , HEWETT JEFFREY W
IPC分类号： C07H21/04 , C07K1/00 , C12Q1/00
CPC分类号： C12Q1/66 , C07K2319/60 , C12N2799/027
摘要： The present invention relates generally to methods to monitor the transport of proteins through the secretory pathway, and methods to monitor ER stress. In particular, the present invention relates to methods to monitor, in real-time, the processing of protein through the secretory pathway, which can be monitored both at a subcellular level by florescence visualization and quantitatively by detecting the secreted luciferase reporter protein. The present invention also relates to methods to assess biological processes in cells, in particular the secretory pathway and ER stress, as well as methods to identify agents which augment or inhibit the secretory pathway and/or ER stress. The present invention also relates to compositions and nucleic constructs encoding a secreted luciferase- fluorescent protein conjugate for methods to monitor protein trafficking in the cell by simultaneous detection of fluorescence and luciferase secretion.
摘要翻译：本发明一般涉及监测蛋白质通过分泌途径的转运的方法，以及监测ER应激的方法。具体而言，本发明涉及实时监测通过分泌途径处理蛋白质的方法，所述分泌途径可以在亚细胞水平上通过荧光可视化以及通过检测分泌的荧光素酶报道蛋白质进行定量监测。本发明还涉及评估细胞中的生物过程，特别是分泌途径和ER应激的方法，以及鉴定增强或抑制分泌途径和/或ER应激的物质的方法。本发明还涉及编码分泌荧光素酶 - 荧光蛋白偶联物的组合物和核酸构建体，用于通过同时检测荧光和萤光素酶分泌来监测细胞中的蛋白质运输的方法。

5. 发明申请

WO2012051622A2 MICROVESICLE-BASED ASSAYS 审中-公开
标题翻译：基于微波的测定
公开(公告)号：WO2012051622A2
公开(公告)日：2012-04-19
申请号：PCT/US2011056589
申请日：2011-10-17
申请人： GEN HOSPITAL CORP , CHEN WALTER , BREAKEFIELD XANDRA O , BALAJ LEONORA , SKOG JOHAN KARL OLOV
发明人： CHEN WALTER , BREAKEFIELD XANDRA O , BALAJ LEONORA , SKOG JOHAN KARL OLOV
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6827 , C12Q1/6883 , C12Q1/6886 , C12Q2600/118 , C12Q2600/156
摘要： Methods are disclosed herein for assaying a biological sample or a bodily fluid obtained from a subject by isolating, obtaining or using a microvesicle fraction from the biological sample or bodily fluid and detecting in the microvesicle fraction the presence or absence of a genetic aberration in an IDH1, IDH2, TP53, PTEN, CDKN2A, NF1, EGFR, RB1, PIK3CA, or BRAF gene. The methods may be used for aiding the diagnosis, prognosis, monitoring, or therapy selection in relation to a disease or other medical condition (e.g., a glioma) in a subject.
摘要翻译：本文公开了用于分析从受试者获得的生物样品或体液的方法，其通过分离，获得或使用来自生物样品或体液的微泡部分并在微泡部分中检测IDH1中存在或不存在遗传畸变，IDH2，TP53，PTEN，CDKN2A，NF1，EGFR，RB1，PIK3CA或BRAF基因。该方法可以用于辅助与受试者中疾病或其他医学病症（例如神经胶质瘤）相关的诊断，预后，监测或治疗选择。

6. 发明申请

WO9955345A9 COMBINATION VIRAL-BASED AND GENE-BASED THERAPY OF TUMORS 审中-公开
标题翻译：组合基于病毒和基因的肿瘤治疗
公开(公告)号：WO9955345A9
公开(公告)日：2000-04-13
申请号：PCT/US9909376
申请日：1999-04-30
申请人： GEN HOSPITAL CORP
发明人： CHIOCCA E ANTONIO , BREAKEFIELD XANDRA O
IPC分类号： A61K35/76 , A61K47/48 , A61K48/00 , C12N7/00 , C12N15/869 , A61K35/00 , C12N15/63
CPC分类号： B82Y5/00 , A61K31/675 , A61K35/763 , A61K38/415 , A61K38/44 , A61K45/06 , A61K47/6891 , A61K48/00 , C12N7/00 , C12N15/86 , C12N2710/16332 , C12N2710/16632 , C12N2710/16643 , C12Y114/14 , C12Y117/04001 , A61K2300/00
摘要： The present invention relates to viral mutants and methods of using these viral mutants for selectively killing neoplastic cells. The viral mutants of the invention are capable of selectively killing neoplastic cells by a combination of viral mediated oncolysis and anti-cancer ("suicide") gene therapy.
摘要翻译：本发明涉及病毒突变体和使用这些病毒突变体选择性杀死肿瘤细胞的方法。本发明的病毒突变体能够通过病毒介导的溶瘤和抗癌（“自杀”）基因治疗的组合选择性地杀死肿瘤细胞。

7. 发明申请

WO0077167A3 HERPES SIMPLEX VIRUS AMPLICON VECTOR TARGETING SYSTEM AND METHOD OF USING SAME 审中-公开
标题翻译： HERPES SIMPLEXVIVUS AMPLICON矢量测向系统及其使用方法
公开(公告)号：WO0077167A3
公开(公告)日：2004-05-06
申请号：PCT/US0016050
申请日：2000-06-12
申请人： GEN HOSPITAL CORP , UNIV CALIFORNIA
发明人： SPEAR MATTHEW A , BREAKEFIELD XANDRA O
IPC分类号： A61K48/00 , C12N15/38 , C12N15/869 , C12N5/10 , C12N15/64 , C12Q1/70
CPC分类号： C12N15/86 , A61K48/00 , C12N2710/16643 , C12N2710/16645 , C12N2810/40 , C12N2810/60
摘要： The present invention relates to herpes simplex virus (HSV) amplicon vectors, and in particular, HSV-1 amplicon vectors, which have been genetically modified and used alone or with consequent genetically modified HSV virus, to target a selected cell type, such as neoplastic cells. The present invention also relates to methods of using such vectors to target a cell, in order to treat a pathologic condition, such as cancer.
摘要翻译：本发明涉及单纯疱疹病毒（HSV）扩增子载体，特别是HSV-1扩增子载体，其已被遗传修饰并且单独使用或随后使用遗传修饰的HSV病毒靶向所选择的细胞类型，例如肿瘤细胞。本发明还涉及使用这些载体靶向细胞的方法，以便治疗诸如癌症的病理状况。

8. 发明申请

WO9324657A3 USE OF GENETIC MARKERS TO DIAGNOSE FAMILIAL DYSAUTONOMIA 审中-公开
标题翻译：使用遗传标记来诊断家族性遗传
公开(公告)号：WO9324657A3
公开(公告)日：1994-02-03
申请号：PCT/US9304946
申请日：1993-05-25
申请人： GEN HOSPITAL CORP
发明人： BLUMENFELD ANAT , GUSELLA JAMES F , BREAKEFIELD XANDRA O
IPC分类号： C07H21/00 , C07H21/04 , C12N20060101 , C12Q20060101 , C12Q1/68
CPC分类号： C12Q1/6858 , C12Q1/6883 , C12Q2600/156 , Y10S435/81
摘要： Familial dysautonomia (FD), the Riley-Day syndrome, is an autosomal recessive disorder characterized by developmental loss of neurons from the sensory and autonomic nervous system. It is limited to the Ashkenazi Jewish population, where the carrier frequency is 1 in 30. We have mapped the FD gene to the chromosome region 9q31-q33 by linkage with ten DNA markers in twenty-six families. The maximum lod score of 21.1 with no recombinants was achieved with D9S58. This marker also showed strong linkage disequilibrium with FD, with one allele present on 73 % of all affected chromosomes compared to 5.4 % of control chromosomes (X2=3142, 15 d.f. p

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