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    • 44. 发明申请
    • STAT3 AS A THERANOSTIC INDICATOR
    • STAT3作为治疗指标
    • WO2008127716A2
    • 2008-10-23
    • PCT/US2008/004813
    • 2008-04-14
    • GEORGE MASON INTELLECTUAL PROPERTIES, INC.LIOTTA, Lance A.PETRICOIN, Emanuel F. IIIWULFKUHLE, Julia
    • LIOTTA, Lance A.PETRICOIN, Emanuel F. IIIWULFKUHLE, Julia
    • G01N33/574
    • G01N33/57415G01N2800/52
    • This invention relates, e.g. , to a method for predicting the response of a subject having estrogen-receptor-positive breast cancer to an inhibitor of the estrogen signaling pathway ( e.g. tamoxifen), comprising measuring in a cancer sample from the subject the level of phosphorylation, compared to a baseline value, of one or more of the following members of an interconnected intracellular signaling pathway: (a) 4EBP 1, and/or (b) p70S6, and/or (c) STAT3, and/or (d) FAK, wherein a significantly elevated level of phosphorylation of 4EBP1, and/or p70S6 and/or STAT3, and/or a significantly decreased level of phosphorylation of FAK, compared to the baseline value, indicates that the subject is likely to be a non-responder to the inhibitor and/or has a poor prognosis. Additional members of the intracellular signaling pathway whose phosphorylation can be measured are also described. Also described is a method for treating breast cancer in a subject in need thereof, wherein the subject exhibits an elevated level of phosphorylation of these markers, comprising administering to the subject an effective amount of one or more inhibitors of members of the interconnected intracellular signaling pathway.
    • 本发明涉及例如用于预测具有雌激素受体阳性乳腺癌的受试者对雌激素信号通路抑制剂(例如他莫昔芬)的反应的方法,包括在受试者的癌症样品中测量磷酸化水平 (a)4EBP1和/或(b)p70S6和/或(c)STAT3和/或(d)与细胞内信号通路相互连接的一个或多个以下成员的基线值; FAK,其中与基线值相比,4EBP1和/或p70S6和/或STAT3的磷酸化水平显着升高和/或FAK的磷酸化水平显着降低,表明受试者可能是非 - 对抑制剂的反应和/或预后差。 还描述了可以测量其磷酸化的细胞内信号传导途径的其他成员。 还描述了一种在有需要的受试者中治疗乳腺癌的方法,其中所述受试者表现出这些标志物的磷酸化水平升高,包括向受试者施用有效量的一种或多种相互连接的细胞内信号传导途径的成员抑制剂 。
    • 45. 发明申请
    • TISSUE PRESERVATION AND FIXATION METHOD
    • 组织保存和固定方法
    • WO2008073187A2
    • 2008-06-19
    • PCT/US2007/022744
    • 2007-10-26
    • GEORGE MASON INTELLECTUAL PROPERTIES, INC.LIOTTA, Lance, A.PETRICOIN, Emanuel, F.GEHO, DavidESPINA, Virginia, A.
    • LIOTTA, Lance, A.PETRICOIN, Emanuel, F.GEHO, DavidESPINA, Virginia, A.
    • A01N1/00G01N1/30
    • G01N1/30A01N1/00
    • This invention relates, e.g ., to a composition that, at room temperature, when contacted with a sample comprising phosphoproteins, can fix and stabilize cellular phosphoproteins, preserve cellular morphology, and allow the sample to be frozen to generate a cryostat frozen section suitable for molecular analysis. The composition comprises ( 1 ) a fixative that is effective to fix the phosphoproteins, and that has a sufficient water content to be soluble for a stabilizer and/or a permeability enhancing agent); (2) a stabilizer, comprising (a) a kinase inhibitor and (b) a phosphatase inhibitor and, optionally, (c) a protease (e.g., proteinase) inhibitor; and (3) a permeability enhancing agent ( e.g. PEG). Methods are described for preserving phosphoproteins, using such a composition. Also described are endogenous surrogate markers for monitoring protein degradation, including the loss of posttranslational modifications (such as phosphorylation), e.g . the following removal of a cell or tissue from a subject; and exogenous molecular sentinels ( e.g . phosphoproteins attached to magnetic nanoparticles) that allow one to evaluate the processing history of a cellular or tissue population sample.
    • 本发明涉及例如在室温下与包含磷蛋白的样品接触时可以固定和稳定细胞磷酸蛋白的组合物,保留细胞形态,并允许样品冷冻以产生适于分子的低温恒温器冷冻部分 分析。 组合物包含(1)固定磷酸蛋白有效的固定剂,并且其具有足够的水含量可溶于稳定剂和/或渗透性增强剂); (2)稳定剂,其包含(a)激酶抑制剂和(b)磷酸酶抑制剂和任选的(c)蛋白酶(例如蛋白酶)抑制剂; 和(3)渗透性增强剂(例如PEG)。 描述了使用这种组合物来保存磷蛋白的方法。 还描述了用于监测蛋白质降解的内源替代标记,包括翻译后修饰(例如磷酸化)的丧失,例如, 以下从受试者中除去细胞或组织; 和外源性分子前哨蛋白(例如连接到磁性纳米颗粒的磷酸蛋白),其允许评价细胞或组织群体样品的加工历史。
    • 46. 发明申请
    • LIVER DISEASE DIAGNOSTIC USING AKT/MTOR/IRS PATHWAY ACTIVATION STATE IN FAT CELLS
    • 在FAT细胞中使用AKT / MTOR / IRS途径激活状态的肝病诊断
    • WO2007136822A2
    • 2007-11-29
    • PCT/US2007/012055
    • 2007-05-21
    • GEORGE MASON INTELLECTUAL PROPERTIES, INC.INOVA HEALTH CARE SERVICESPETRICOIN, EmanuelLIOTTA, LanceCALVERT, ValerieYOUNOSSI, Zobair
    • PETRICOIN, EmanuelLIOTTA, LanceCALVERT, ValerieYOUNOSSI, Zobair
    • G01N33/68G01N33/573G01N33/00
    • G01N33/6893G01N2800/085
    • This invention relates, e.g. , to a method to distinguish NASH (non-alcoholic steatohepatitis) from non-NASH types of NAFLD (non-alcoholic fatty liver disease), comprising (a) determining the phosphorylation state of one or more members of the AKT/mTOR/IRS pathway ( e.g. , IRS-1 (S612), AKT (S308), FKHR (T24), FKHRL (T32), GSK3 (S21/9), EIF4G (S1108), SHC (Y317), PKC-delta (T505), PKA (T197), CREB (S133), and/or FAK (Y397) in adipose tissue from a subject, wherein a significantly altered level of phosphorylation of the one or more members compared to a suitable reference standard indicates that the subject is likely to have NASH; and/or (b) determining the phosphorylation state of FAK (focal adhesion kinase) in adipose tissue from a subject, wherein a significantly increased level of phosphorylation in FAK compared to a suitable reference standard indicates that the subject is likely to have NASH. Treatment modalities based on this diagnostic method, and kits for carrying out a diagnostic method or a treatment method are also disclosed.
    • 本发明涉及例如将NASH(非酒精性脂肪性肝炎)与非NASH类型的NAFLD(非酒精性脂肪性肝病)区分开的方法,其包括(a)确定AKT中一种或多种成员的磷酸化状态 / mTOR / IRS途径(例如IRS-1(S612),AKT(S308),FKHR(T24),FKHRL(T32),GSK3(S21 / 9),EIF4G(S1108),SHC(Y317) (T505),PKA(T197),CREB(S133)和/或FAK(Y397),其中一个或多个成员的磷酸化水平与合适的参考标准相比显着改变的磷酸化水平表明, 受试者可能具有NASH;和/或(b)确定来自受试者的脂肪组织中FAK(粘着斑激酶)的磷酸化状态,其中与合适的参考标准相比,FAK中磷酸化水平的显着增加水平表明该受试者 可能有NASH。基于这种诊断方法的治疗方式,以及进行诊断的试剂盒 还公开了方法或处理方法。
    • 50. 发明申请
    • IMPROVED ONE-STEP CELL AND TISSUE PRESERVATIVE FOR MORPHOLOGIC AND MOLECULAR ANALYSIS
    • 改进的一步细胞和组织形态学和分子分析保守剂
    • WO2011091438A1
    • 2011-07-28
    • PCT/US2011/022463
    • 2011-01-25
    • GEORGE MASON INTELLECTUAL PROPERTIES, INC.ESPINA, VirginiaLIOTTA, Lance, A.MUELLER, Claudius
    • ESPINA, VirginiaLIOTTA, Lance, A.MUELLER, Claudius
    • A01N1/02G01N33/48
    • G01N1/30A01N1/00
    • The invention relates to a one-step chemical composition that preserves animal tissue, cells, and biomolecules, such as human tissue, human cells, and biomolecules therein. It improves the fidelity and morphologic structure of cells, organelles, and nuclear chromatin, and maintains and enhances the cellular antigenicity for immunohistochemistry and flow cytometry, while preserving proteins, post-translational modifications of proteins, and nucleic acids. In one embodiment, the composition comprises a) a non-aldehyde precipitating fixative at a concentration below 25% (volume/volume), b) a reversible/cleavable protein cross-linker that targets lipid-associated molecules, and c) a c reversible/cleavable protein cross-linker that targets water soluble molecules. In another embodiment, the composition further includes a kinase inhibitor, a phosphatase inhibitor, and a permeation enhancer. In still another embodiment, the compositions further include lactic acid at a concentration sufficient to maintain cellular nuclear volume at a level equivalent to aldehyde fixation of the same type of cell. In a further embodiment, the composition comprises: a) a precipitating fixative, b) a reversible/cleavable cross-linker, c) a permeation enhancer, d) a kinase inhibitor, e) a phosphatase inhibitor, and f) a carboxylic acid. In a still further embodiment, the invention comprises method for preserving a biological sample by contacting the sample with the composition of the invention under conditions effective for the preservation of the sample.
    • 本发明涉及保留动物组织,细胞和生物分子的一步化学组合物,例如人组织,人细胞和生物分子。 它提高细胞,细胞器和核染色质的保真度和形态结构,并保持和增强免疫组织化学和流式细胞术的细胞抗原性,同时保留蛋白质,蛋白质的翻译后修饰和核酸。 在一个实施方案中,组合物包含a)浓度低于25%(体积/体积)的非醛沉淀固定剂,b)靶向脂质相关分子的可逆/可切割蛋白质交联剂,以及c)可逆可逆/ 可裂解的蛋白质交联剂,其靶向水溶性分子。 在另一个实施方案中,组合物还包含激酶抑制剂,磷酸酶抑制剂和渗透增强剂。 在另一个实施方案中,组合物还包括浓度足以将细胞核体积保持在与相同类型细胞的醛固定相当的水平的乳酸。 在另一个实施方案中,组合物包括:a)沉淀固定剂,b)可逆/可切割交联剂,c)渗透促进剂,d)激酶抑制剂,e)磷酸酶抑制剂,和f)羧酸。 在又一个实施方案中,本发明包括通过在有效保存样品的条件下使样品与本发明的组合物接触来保存生物样品的方法。