PHASE IB STUDY OF BRYOSTATIN 1--NSC339555
Steven Grant, Professor
Virginia Commonwealth University Po Box 980568 Richmond, Va 232980568
Grant 2M01RR000065-350457 from National Center For Research Resources
Abstract: The goal is to determine whether bryostatin 1 administered to patients with advanced malignancies is capable of inducing significant and sustained down-regulation of total PKC activity in a surrogate target tissue. The second major goal is to determine whether in vivo administration of bryostatin results in plasma bryostatin 1 concentrations mimicking those reported to potentiate ara-C related apoptosis in leukemic cells in vitro.
Keywords: bryostatin, clinical trial phase I, drug screening /evaluation, human therapy evaluation, neoplasm /cancer chemotherapy, antileukemic agent, cell death, enzyme activity, neoplastic cell, outcomes research, protein kinase C, clinical research, human subject
Project start date: 1997-01-20
Project end date: 1997-11-30
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to Steven Grant
BRYOSTATIN 1 (NSC 339555) And FLUDARABINE IN LYMPHOCYTIC LEUKEMIA
Steven Grant, Professor
Virginia Commonwealth University Po Box 980568 Richmond, Va 232980568
Grant 5M01RR000065-390552 from National Center For Research Resources
Abstract: To determine the toxicities and appropriate doses for the combination of bryostatin and fludarabine in the treatment of chronic lymphocytic leukemia and indolent lymlphoma
Keywords: acute lymphocytic leukemia, bryostatin, drug screening /evaluation, lymphoma, neoplasm /cancer chemotherapy, clinical trial phase I, dosage, drug adverse effect, human therapy evaluation, clinical research, human subject
Project start date: 2000-12-01
Project end date: 2001-11-30
BRYOSTATIN 1 (NSC 339555) & FLUDARABINE IN LYMPHOCYTIC LEUKEMIA
Steven Grant, Professor
Virginia Commonwealth University Po Box 980568 Richmond, Va 232980568
Grant 3M01RR000065-37S10552 from National Center For Research Resources
Abstract: To determine the toxicities and appropriate doses for the combination of bryostatin and fludarabine in the treatment of chronic lymphocytic leukemia and indolent lymlphoma
Keywords: acute lymphocytic leukemia, bryostatin, drug screening /evaluation, lymphoma, neoplasm /cancer chemotherapy, clinical trial phase I, dosage, drug adverse effect, human therapy evaluation, clinical research, human subject
Project start date: 1998-12-01
Project end date: 1999-11-30
Flavopiridol / Bortezomib In Myeloma / B-Cell Neoplasms
Steven Grant, Professor
Virginia Commonwealth University Po Box 980568 Richmond, Va 232980568
Grant 5R21CA110953-02 from National Cancer Institute IRG: CONC
Abstract: The results of recent studies indicate that neoplastic cells, particularly those of hematopoietic origin, are exquisitely sensitive to apoptosis induced by simultaneous disruption of survival signaling and cell cycle regulatory pathways. Bortezomib, the prototypical clinically relevant proteasome inhibitor, has shown impressive activity against multiple myeloma refractory to conventional therapies, as well as mantle cell lymphoma (MCL) and indolent NHL. Flavopiridol (NSC 649890), the first cyclin-dependent kinase inhibitor to enter clinical trials, induces apoptosis in malignant human hematopoietic cells at pharmacologically achievable concentrations. Recent preclinical studies from our laboratory indicate that co-administration of bortezomib and flavopiridol at low concentrations results in a highly synergistic induction of mitochondrial damage and apoptosis in human leukemia and myeloma cell lines as well as in primary patient-derived specimens. These events are associated with multiple perturbations in signaling and survival pathways, including activation of the stress-related JNK cascade, down-regulation of the anti-apoptotic proteins Mcl-1 and XlAP, and inactivation of the cytoprotective NF-kB pathway. The hypothesis that similar events will occur in myeloma and lymphoma cells exposed to these agents in vivo has prompted the development of a Phase I trial of bortezomib and flavopiridol in patients with multiple myeloma, MCL, and indolent B-cell NHL. The goals of this project are (1) To conduct a Phase I trial of flavopiridol and bortezomib administered as bolus IV infusions BIW (days 1 and 4, 8 and 11) q 3 wks to determine the maximally tolerated drug doses for this schedule; (2) To define dose-limiting toxicities; (3) To assess preliminary clinical activity of this regimen in patients with myeloma and B-cell neoplasms; (4) To conduct correlative laboratory and PK studies to test the hypothesis that in vivo administration of bortezomib and flavopiridol will result in NF-kB inactivation, JNK activation, and down-regulation of Mcl-1 and XIAP in multiple myeloma cells.
Keywords: B cell lymphoma, drug screening /evaluation, flavopiridol, human therapy evaluation, multiple myeloma, neoplasm /cancer chemotherapy, protein kinase inhibitor, clinical trial phase I, drug administration rate /duration, drug adverse effect, neoplasm /cancer pharmacology, nonHodgkin s lymphoma, nuclear factor kappa beta, pharmacokinetics, phosphorylation, SDS polyacrylamide gel electrophoresis, clinical research, fluorescence microscopy, human subject, patient oriented research, western blotting
Project start date: 2004-06-15
Project end date: 2008-05-31
5R21CA110953-02 (2005): $213629
1R21CA110953-01 (2004): $231985
PROMOTION OF NUCLEOSIDE ANALOG INDUCED APOPTOSIS BY HDIS
Steven Grant, Professor
Virginia Commonwealth University, Po Box 980568, Richmond, Va 23298-0568
Grant 5R01CA063753-14 from National Cancer Institute
Abstract: The goal of this renewal application is to develop a rational basis for employing novel, clinically relevant histone deacetylase inhibitors (HDIs), particularly the benzamide HDI MS-275 as well as SAHA and LAQ824, to enhance the antileukemic potential of fludarabine and other established nucleoside analogs. In the preceding funding period, the mechanism underlying synergistic interactions between the macrocyclic lactone bryostatin 1 or the PKC inhibitor UCN-01 and various antileukemic agents were determined to be elaboration of TNFalpha or inhibition of Chk1 rather than down-regulation/inhibition of protein kinase C; moreover, several clinical trials combining bryostatin 1 or UCN-01 and nucleoside analogs have since been initiated. Subsequently, it has been determined that antileukemic interactions between HDIs and nucleoside analogs such as fludarabine are, if anything, more pronounced than those involving bryostatin 1. Furthermore, such interactions have been related to several recently described pro-apoptotic HDI actions, including a) interruption of the cytoprotective MEK1/2/ERK and Akt modules accompanied by JNK activation; b) enhanced generation of reactive oxygen species (ROS); and c) perturbations in the expression of certain anti-apoptotic (i.e., Mcl-1 and XIAP) and cell cycle (i.e., cyclin D1, p27KIP1)proteins. Significantly, MS-275 treatment markedly increases fludarabine-mediated generation of ceramide, a pro-apoptotic lipid second messenger linked to nucleoside analog-mediated lethality in leukemic cells. The specific aims of this proposal are to 1) define the functional roles of inactivation of survival (e.g., ERK, Akt and stress-related (e.g., JNK) pathways by HDIs in synergistic interactions with fludarabine; 2) clarify the role of enhanced ROS generation by HDIs in these events; 3) characterize the functional role of Mcl-1, XlAP, p27KIP1, and cyclin D1 down-regulation in HDI/fludarabine interactions; 4) determine whether enhanced generation of ceramide and possibly reciprocal changes in levels of sphingosine-1-phosphate play functional roles in antileukemic synergism; 5) extend these findings to primary human leukemia cells and normal hematopoietic progenitors to determine whether a basis for therapeutic selectivity exists. Information derived from these studies may provide a rational foundation for new therapeutic approaches in leukemia in which novel HDIs such as MS-275 are combined with fludarabine or other established nucleoside analogs
Keywords: 1-(2-Oxo-4-amino-1, 2-dihydropyrimidin-1-yl)-2-deoxy-2, 2-difluororibose; 2`, 2`-DFDC; 2`, 2`-difluoro-2`-deoxycytidine; 2`, 2`-difluorodeoxycytidine; 2`-deoxy-2`-difluorocytidine; 2`Deoxy-2`, 2`-Difluorocytidine; 2, 2 difluorodexoycytidine; 2-Fluoro-9-beta-arabinofuranosyladenine; 2-Fluorovidarabine; 2-fluoroadenine arabinoside; 9-Beta-D-arabinofuranosyl-2-fluoro-9H-purin-6-amine; 9-Beta-D-arabinofuranosyl-2-fluoroadenine; API3; Active Oxygen; Acute leukemia; Apoptosis; Apoptosis Pathway; Apoptotic; Ara-C; BIRC4; BIRC4 gene; Benzamides; Blood (Leukemia); Bryostatin-1; CCND1 Protein; CDK inhibitor p27; CDKN1B; CDKN1B protein; CDKN4 protein; Calcium Phospholipid-Dependent Protein Kinase; Calcium-Activated Phospholipid-Dependent Kinase; Cell Communication and Signaling; Cell Cycle; Cell Death, Programmed; Cell Division Cycle; Cell Signaling; Cells; Ceramide (lipids); Ceramides; Chemosensitization; Chemosensitization/Potentiation; Class; Clinic; Clinical Trials; Clinical Trials, Unspecified; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p27; Cyclins; Development; Difluorodeoxycytidine; Down-Regulation; Down-Regulation (Physiology); Downregulation; Drug Potentiations; Event; F Ara A; Family member; Foundations; Funding; G1/S-Specific Cyclin D1; Generations; Goals; Grant; HDAC Agent; Hematologic Cancer; Hematologic Malignancies; Hematologic Neoplasms; Hematological Malignancies; Hematological Neoplasms; Hematological Tumor; Hematopoietic; Hematopoietic Cancer; Histone Deacetylase Inhibitor; Human; Human, General; ILP; Induction of Apoptosis; Interruption; Intracellular Communication and Signaling; Intracellular Second Messengers; Investigators; JNK; JNK1; JNK1A2; JNK21B1/2; Kip1 protein; LAQ824; LEUKCL; Lactone Compound; Lactones; Leukemia, Granulocytic; Leukemias, General; Leukemic Cell; Link; Lipids; Lymphoid; MAP Kinase 8 Gene; MAP-ERK Kinase; MAPK ERK Kinases; MAPK8; MAPK8 gene; MEKs; MIHA; MS-275; Malignant Hematologic Neoplasm; Man (Taxonomy); Man, Modern; Mediating; Molecular; Myelocytic Leukemia; Myelogenous Leukemia; Myeloid Leukemia; NVP-LAQ824; Non-Lymphoblastic Leukemia; Non-Lymphocytic Leukemia; Normal Cell; Numbers; Output; Oxygen Radicals; P27KIP1; PKC; PRAD1 Protein; PRKM8; Pathway interactions; Phase I/II Trial; Phospholipid-Sensitive Calcium-Dependent Protein Kinase; Play; Potentiation; Potentiations, Drug; Pro-Oxidants; Protein Kinase C; Protein Kinase C Inhibitor; Proteins; Proto-Oncogene Proteins c-bcl-1; Reactive Oxygen Species; Research Personnel; Researchers; Role; SAHA; SAPK1; Second Messenger Systems; Second Messengers; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Signal Transduction Systems; Signaling; Stress; Suberoylanilide Hydroxamic Acid; Testing; Therapeutic; Translating; Translatings; Vorinostat; XIAP; analog; antileukemic agent; base; bcl-1 Proto-Oncogene Products; bcl-1 Proto-Oncogene Proteins; bcl1 Proto-Oncogene Proteins; biological signal transduction; bryostatin; c-bcl-1 Proteins; clinical investigation; clinical relevance; clinically relevant; cyclin D; cyclin-dependent kinase inhibitor 1B; dFdC; dFdCyd; fludarabine; gemcitabine; gene product; language translation; leukemia; myeloid granulocytic leukemia; myelosis; new approaches; new therapeutics; next generation therapeutics; novel; novel approaches; novel strategies; novel strategy; novel therapeutics; nucleoside analog; p27 Kip1 protein; p27 protein; p27(Kip); p27-Kip1; p27Kip1 protein; pathway; progenitor; second messenger; social role; sphingosine 1-phosphate; stem; suberanilohydroxamic acid; synergism
Project start date: 1994-06-01
Project end date: 2010-04-30
Budget start date: 1-MAY-2008
Budget end date: 30-APR-2010
5R01CA063753-14 (2008): $0
5R01CA063753-13 (2007): $256009
5R01CA063753-12 (2006): $263655
5R01CA063753-11 (2005): $267470
2R01CA063753-10 (2004): $270000
CHECKPOINT ABROGATION AND MEK 1/2 INHIBITION IN MYELOMA
Steven Grant, Professor
Virginia Commonwealth University, Po Box 980568, Richmond, Va 23298-0568
Grant 5R01CA100866-06 from National Cancer Institute
Abstract: The goal of this renewal application is to define the largely unexplored relationship between the ATM/ATR/Chk1 DNA-damage checkpoint machinery and the Src/Ras/Raf/MEK/ERK survival signaling pathway in multiple myeloma (MM) cells, and to exploit this knowledge therapeutically. The previous project stemmed from the observation that the multi-kinase and Chk1 inhibitor UCN-01 triggered MEK/ERK activation in MM cells, and that pharmacologic interruption of this pathway strikingly induced apoptosis in malignant cells, including primary CD138+, but not normal cells. During the preceding period, important new insights emerged, including the observations that a) disruption of Chk1 by UCN-01 activates MEK1/2/ERK1/2 through a Ras- dependent mechanism that limits lethality; b) these findings may represent a generalized phenomenon involving novel and more specific Chk1 inhibitors as well as new agents acting upstream of Chk1 (i.e., ATM/ATR inhibitors); c) interrupting the MEK/ERK pathway at various levels, e.g., by new, clinically relevant MEK1/2 inhibitors as well as antagonists of upstream signaling targets (e.g., Src and Ras) dramatically potentiates checkpoint abrogator lethality; d) the lethality of these regimens is associated with marked potentiation of DNA damage and induction of Bim-dependent apoptosis; e) this approach is highly lethal toward non-cycling G0G1 MM cells and circumvents Mcl-1-related resistance; and e) this strategy is effective in vivo. Collectively, our findings argue that in MM, activation of the Ras/Raf/MEK/ERK pathway represents a critical cytoprotective response to Chk1/ATM/ATR inhibitors, and that blockade of the former pathway leads to pronounced activity against MM cells both in vitro and in vivo. The specific aims of this proposal are to 1) validate, through genetic means, the Chk1 and MEK/ERK pathways as targets for regimens combining new generation Chk1 and ATM/ATR inhibitors with MEK/ERK pathway antagonists in MM, and investigate mechanisms responsible for synergistic interactions, focusing on Bim upregulation, DNA damage induction, and cdc2 activation; 2) test the activity and selectivity of novel regimens toward primary CD138+ MM cells, and elucidate mechanisms responsible for the striking susceptibility of non-cycling G0G1 and primary MM cells to this strategy; 3) define the basis by which these regimens circumvent cytoprotection by Mcl-1, a critical MM survival factor, and use genetic and pharmacologic strategies to determine whether disabling of Bcl-2/Bcl-xL improves therapeutic activity further; and 4) determine whether regimens combining second-generation Chk1 and ATM/ATR inhibitors with MEK/ERK pathway antagonists overcome conventional, novel (e.g., stromal-cell related), and bortezomib resistance, emphasizing in vivo flank and systemic xenograft MM model systems. These studies will establish a new treatment paradigm for Chk1 inhibitors and lay the foundation for one or more novel Phase I trials in refractory MM in which emerging inhibitors of the ATM/ATR/Chk1 checkpoint machinery and the MEK/ERK survival pathway are rationally combined to trigger MM cell death. Despite recent advances in the treatment of multiple myeloma, it largely remains an incurable disease, and new treatment options are urgently needed. The goal of this study is to develop an entirely novel therapeutic strategy for the treatment of this disorder involving agents that interrupt DNA damage checkpoints and an important survival signaling pathway. If successful, these studies could lead to the development of a more effective therapy for patients with refractory multiple myeloma and potentially other blood cancers
Keywords: Apoptosis; Apoptosis Pathway; Biological Models; Bortezomib; CDC2 Protein Kinase; CDK1; Cancer Treatment; Cell Communication and Signaling; Cell Cycle Controller cdc2; Cell Death; Cell Death, Programmed; Cell Division Control Protein 2 Homolog; Cell Division Cycle 2; Cell Division Cycle 2 Protein; Cell Protection; Cell Signaling; Cells; Checkpoint kinase 1; Chemosensitization; Chemosensitization/Potentiation; Clinical Trials; Clinical Trials, Phase I; Clinical Trials, Unspecified; Cyclin-Dependent Kinase 1; Cytoprotection; DNA Damage; DNA Injury; DNA damage checkpoint; DNA damage checkpoint response; DNA damage response, signal transduction resulting in cell cycle arrest; Development; Disease; Disorder; Early-Stage Clinical Trials; Employee Strikes; Foundations; Funding; Generations; Genetic; Goals; Hematopoietic Cell Tumor; Hematopoietic Malignancies; Hematopoietic Neoplasms; Hematopoietic Neoplasms including Lymphomas; Hematopoietic Tumor; Hematopoietic and Lymphoid Cell Neoplasm; Hematopoietic and Lymphoid Neoplasms; Hematopoietic, Including Myeloma; Heterograft; Human; Human, General; In Vitro; Interruption; Intracellular Communication and Signaling; Knowledge; Laboratories; Lead; Link; MAP-ERK Kinase; MAPK ERK Kinases; MEKs; Malignant Cell; Malignant Hematopoietic Neoplasm; Malignant Neoplasm Therapy; Malignant Neoplasm Treatment; Mammals, Mice; Man (Taxonomy); Man, Modern; Mice; Model System; Modeling; Models, Biologic; Multiple Myeloma; Murine; Mus; Myeloma, Plasma-Cell; New Agents; Normal Cell; Pathway interactions; Patients; Pb element; Phase 1 Clinical Trials; Phase I Clinical Trials; Phase I Study; Potentiation; Predisposition; Process; Radiation; Ras/Raf; Refractory; Regimen; Resistance; Signal Pathway; Signal Transduction; Signal Transduction Systems; Signaling; Staging; Strikes; Strikes, Employee; Stromal Cells; Susceptibility; Testing; Therapeutic; Transplantation, Heterologous; Up-Regulation; Up-Regulation (Physiology); Upregulation; Xenograft; Xenograft procedure; Xenotransplantation; anticancer therapy; base; biological signal transduction; blood cancer; cancer cell; cancer therapy; cdc2 gene product; cdc2+ Protein; cdk1 Kinase; chk1 kinase; chk1 protein kinase; clinical investigation; clinical relevance; clinically relevant; disease/disorder; effective therapy; heavy metal Pb; heavy metal lead; improved; in vivo; inhibitor; inhibitor/antagonist; insight; myeloma; myelomatosis; necrocytosis; new therapeutics; next generation therapeutics; novel; novel therapeutics; p34 (cdc2); p34 Protein Kinase; p34CDC2; pathway; phase 1 study; phase 1 trial; phase I trial; pro-apoptotic protein; protocol, phase I; public health relevance; ray (radiation); resistant; response; small molecule; stem; treatment strategy
Relevance: Despite recent advances in the treatment of multiple myeloma, it largely remains an incurable disease, and new treatment options are urgently needed. The goal of this study is to develop an entirely novel therapeutic strategy for the treatment of this disorder involving agents that interrupt DNA damage checkpoints and an important survival signaling pathway. If successful, these studies could lead to the development of a more effective therapy for patients with refractory multiple myeloma and potentially other blood cancers
Project start date: 2003-05-01
Project end date: 2014-04-30
Budget start date: 1-MAY-2010
Budget end date: 30-APR-2011
PFA/PA: PA-07-070
5R01CA100866-06 (2010): $248557
Sponsored Links Excellgen http://Excellgen.com
2R01CA100866-05A2 (2009): $249983
POTENTIATION OF TAXANE ACTIVITY BY MEK/MAPK INHIBITORS
Steven Grant, Professor
Virginia Commonwealth University Po Box 980568 Richmond, Va 232980568
Grant 5R01CA083705-04 from National Cancer Institute IRG: ET
Abstract: The goal of this proposal is to elucidate the mechanism(s) underlying recent observations that inhibitors of the MEK/MAPK signal transduction pathway a) markedly potentiate paclitaxel- and taxotere-induced lethality toward leukemic cells in a sequence-dependent manner and b) circumvent resistance coffered by Bcl-2/Bcl-xL-mediated blockade of the cell death pathway. We will test the hypothesis that these phenomenon involve perturbations in stress-survival signaling, post-translational modification of anti-apoptotic proteins (Bcl-2/Bcl-xL), or cell cycle dysregulation, (e.g., promotion of mitotic arrest and/or disruption of the mitotic spindle checkpoint). In Aim #1, we will relate enhancement of taxane-mediated apoptosis by subsequent exposure to selective MEK/MAPK inhibitors (PD98059, U0126, and SL327) and agents that interrupt the PKC/MAPK axis (e.g., bryostatin 1, CGP41251) to specific alterations in JNK/MAPK signaling and G2M arrest events, particular CDK1 activation. In Aim #2, an inducible Raf-1 activation system will be used to test functionally the hypothesis that such inhibitors enhance paclitaxel by inhibiting the Raf-1 downstream targets MEK1/2 and MAPK. In Aim #3, cells over- expressing actions of MEK/MAP kinase inhibitors involve modulation of Bcl-2/Bcl-xL phosphorylation status and/or a diminution in the threshold of taxane-induced mitochondrial dysfunction. In Aim #4, the hypotheses that MEK/MAP kinase inhibitors act by antagonizing CDK1 activation (thereby promoting mitotic arrest) or, alternatively, induction of the cyclin-dependent kinase inhibitor p21/CIP1 in taxane-pretreated cells will be tested utilizing the CDK1 inhibitor butyrolactone I and p21/CIP1 antisense-expressing cell lines respectively. Finally, comparisons will be made between the effects of MEK/MAPK inhibitors on taxane-induced lethality toward normal (VFU-GM, HPP-CFC) versus leukemic (L-CFU) progenitors to identify a possible basis for therapeutic selectivity. This information could provide a rationale for using novel MEK/MAP kinase inhibitors with in vivo activity to enhance the anti-tumor activity of taxanes in hematological and ultimately non-hematological malignancies.
Keywords: antineoplastic, biological signal transduction, enzyme inhibitor, mitogen activated protein kinase, paclitaxel, pharmacokinetics, BCL2 gene /protein, acute lymphocytic leukemia, apoptosis, butyrolactone, cell cycle, cell growth regulation, cyclin dependent kinase, cytochrome c, drug resistance, neoplasm /cancer pharmacology, neoplastic cell, neoplastic growth, oncoprotein p21, phosphorylation, posttranslational modification, human tissue, myeloid stem cell, tissue /cell culture
Project start date: 2000-01-10
Project end date: 2003-12-31
5R01CA083705-04 (2003): $168863
5R01CA083705-02 (2001): $160466
1R01CA083705-01 (2000): $157981
MODULATION OF ARA-C INDUCED APOPTOSIS BY BRYOSTATION 1
Steven Grant, Professor
Virginia Commonwealth University Po Box 980568 Richmond, Va 232980568
Grant 5R01CA063753-04 from National Cancer Institute IRG: ET
Abstract: The goal of this proposal is to elucidate the mechanism by which the protein kinase C (PC-C) activator, bryostatin 1, as well as other agents acting through the PK-C signal transduction pathway, enhance the ability of 1-Beta-D-arabinofuranosylcytosine (ara-C) to induce apoptosis (programmed cell death; PCD) in myeloid leukemia cells, resulting in synergistic antileukemic effects for the combination. This approach is based on the hypothesis that basal activity of PK-C protects myeloid leukemia cells from PCD, and that down-regulation of the enzyme, or of one or more of its isoforms, sensitizes cells to drug-induced apoptosis. Initially, the dose and schedule-dependent effects of PK-C activators, both physiologic (e.g., phospholipase C, diacyglycerol) and non- physiologic (e.g., bryostatin 1, PDBu, mezerein), as well as inhibitors (e.g., H07, staurosporine, calphostin, chelerythrine, gossypol, hypericin), will be fully characterized with respect to ara-C-induced endonucleolytic DNA cleavage, apoptotic morphology, and inhibition of clonogenicity in HL-60 cells and other leukemia cell lines (e.g., U937). The total activity and subcellular distribution (nuclear, cytoplasmic, and membrane) of PK-C will be monitored in parallel, and correlations sought between specific perturbations and potentiation (or antagonism) of ara-C-induced apoptosis. Northern analysis, immunoblotting techniques, and isoform assays will be employed to determine whether alterations in the activity (e.g., down-regulation) of individual PK-C isoforms (e.g., alpha, Beta, or gamma) are specifically related to augmentation of ara-C actions. Perturbations in PK-C activity associated with potentiation of apoptosis will be assessed with respect to the expression of oncogenes (e.g., c-jun, bcl-2, c-myc) implicated in this process; these associations may subsequently be evaluated more definitely through the use of antisense blockade strategies and dominant-negative transfectants. Attempts will be made to determine whether agents acting through signal transduction pathways also potentiate ara-C-induced apoptosis in primary AML cell cultures, and whether this strategy selectively spares normal primitive human hematopoietic progenitors exhibiting stem-cell characteristics (e.g., HPP-CFC). It is anticipated that these studies will provide a rational basis for employing bryostatin 1 and other agents acting through second messenger pathways to improve the antileukemic efficacy and selectivity of ara-C and possibly other nucleoside analogs.
Keywords: antileukemic agent, biological signal transduction, bryostatin, cytosine arabinoside, myelogenous leukemia, programmed cell death, protein kinase C, chemical cleavage, chemical kinetics, enzyme activity, hematopoietic stem cell, oncogene, protein isoform, second messenger, staurosporine, northern blotting, tissue /cell culture, transfection, western blotting
Project start date: 1994-06-01
Project end date: 1998-06-30
5R01CA063753-04 (1997): $1
5R01CA063753-02 (1995): $1
Flavopiridol And Differentiation-inducers In Leukemia
Steven Grant, Professor
Virginia Commonwealth University Po Box 980568 Richmond, Va 232980568
Grant 5R01CA093738-04 from National Cancer Institute IRG: ET
Abstract: The goal of this proposal is to develop a rational basis for a novel antileukemic strategy combining pharmacologic cyclin-dependent kinase (CDK) inhibitors with clinically relevant differentiation-inducers (DIs). This approach has been prompted by the discovery that flavopiridol (FP; NSC 649890), a potent CDK ATP-binding domain inhibitor, fails to lower the maturation threshold for human leukemia cells (e.g., U937, HL-60) despite promoting cell cycle arrest; instead, it interacts synergistically with a variety of DIs, e.g., PMA, bryostatin 1, and multiple histone deacetylase inhibitors (HDIs), including sodium butyrate (SB) and SAHA, to trigger mitochondrial damage and apoptosis. Combination of FP with DIs is associated with dysregulation of multiple signaling and cell cycle regulatory pathways, including interference with the induction of the endogenous CDK inhibitor p21 CIPI, degradation of p27KIP1, accelerated dephosphorylation and proteolysis of pRb, derepression of E2F, and enhanced activation of p42/44 MAPK. The specific aims of this proposal are (1) to test the hypothesis, using p21 CIP1 antisense, nuclear localization signal and CDK binding domain mutants, as well as inducible constructs, that dysregulation of this CDKI contributes functionally to FP/DI-induced apoptosis; (2) to determine what role, if any, dysregulation of pRb dephosphorylation, cleavage, and E2F activation play in potentiation of DI-associated apoptosis by FP; and (3) to investigate, using stable transfectants ectopically expressing Bcl-2/Bcl-xL as well as phosphorylation-deficient mutants, mechanisms by which FP and DIs circumvent the block to mitochondrial damage and apoptosis conferred by these proteins. For each of these studies, stable cell lines inducibly expressing Raf-1 or MEK1 will be employed to define the role of the PKC/Raf/MEKIMAPK cascade in FP/DI-mediated lethality. Lastly, the effect of combinations of FP with various DIs will be compared in primary human leukemic blasts and normal progenitors (e.g., CD34+, DR-, 71-) to establish whether a basis for therapeutic selectivity exists. It is hoped that information derived from this proposal will lay the foundation for the implementation of a novel therapeutic strategy combining pharmacologic CDK inhibitors with DIs in the treatment of leukemia and possibly other hematologic malignancies.
Keywords: cell differentiation, cyclin dependent kinase, enzyme inhibitor, flavopiridol, leukemia, oncoprotein p21, apoptosis, biological signal transduction, mitogen activated protein kinase, protein localization, cell line, clinical research, human subject, transfection
Project start date: 2002-01-01
Project end date: 2007-07-31
5R01CA093738-04 (2005): $249375
5R01CA093738-03 (2004): $249375
5R01CA093738-02 (2003): $249375
Checkpoint Abrogation And MEK1/2 Inhibition In Myeloma
Steven Grant, Professor
Virginia Commonwealth University Po Box 980568 Richmond, Va 232980568
Grant 5R01CA100866-04 from National Cancer Institute IRG: ET
Abstract: The goal of this proposal is to develop a novel therapeutic strategy for the treatment of multiple myeloma employing checkpoint abrogators such as UCN-01 in combination with inhibitors of the MEK1/2/MAP kinase pathway. It is based upon the recent discovery that exposure of multiple myeloma and other malignant hematopoietic cells to UCN-01 increases activity of p34cdc2(cdk1), an event associated with MAP kinase activation. Furthermore, interruption of the latter process (e.g., by pharmacologic MEK inhibitors such as U0126 or PD184352) results in a dramatic increase in mitochondrial injury, caspase activation, and apoptosis. Significantly, UCN-01/MEK inhibitor-mediated lethality toward myeloma cells is undiminished by conventional or cell adhesion-related drug resistance mechanisms, or by exogenous survival-related cytokines such as IL-6 and IGF-1. Finally, consistent with evidence that neoplastic cells are selectively impaired in checkpoint control, primary CD138+ bone marrow-derived human myeloma cells appear to be significantly more sensitive than their normal counterparts to this novel strategy. The specific aims of this proposal are a) to elucidate the mechanism(s) by which UCN-01 and MEK inhibitors interact synergistically in myeloma cells, focusing on the functional significance of perturbations in signaling/cell cycle regulatory pathways (e.g., p34cdc2, Raf/MEK/MAP kinase, NFkB, p27KIP1, and cyclin D1; b) to test the hypothesis that the anti-myeloma activity of the UCN-01/MEK inhibitor regimen can be further enhanced by coadministration of proteasome inhibitors, possibly by sparing the cdc25C phosphatase; c) to test the hypothesis that the clinically relevant geldanamycin analog 17-AAG interacts synergistically with UCN-01 in myeloma cells by mimicking the ability of MEK inhibitors to disrupt the cytoprotective MAP kinase cascade; and d) to establish whether these regimens exert selective toxicity toward primary human myeloma cells while sparing their normal hematopoietic counterparts. It is anticipated that information derived from this proposal will serve as a foundation for initiating Phase I trials combining checkpoint abrogators such as UCN-01 with pharmacologic MEK inhibitors in the treatment of multiple myeloma and possibly other hematologic malignancies. Such studies may also provide a paradigm for the development of a novel therapeutic strategy in which inhibitors of cell cycle regulatory and survival signal transduction pathways are rationally combined in the treatment of cancer.
Keywords: apoptosis, cell cycle protein, cell growth regulation, drug interaction, enzyme inhibitor, mitogen activated protein kinase, multiple myeloma, pharmacokinetics, biological signal transduction, combination chemotherapy, cytoprotection, cytotoxicity, drug resistance, enzyme activity, enzyme induction /repression, enzyme therapy, mitochondrial membrane, neoplasm /cancer chemotherapy, cell line, clinical research, flow cytometry, gel mobility shift assay, human tissue, immunoprecipitation, transfection, western blotting
Project start date: 2003-05-01
Project end date: 2008-04-30
5R01CA100866-04 (2006): $243515
Sponsored Links Excellgen http://Excellgen.com
5R01CA100866-03 (2005): $249375
5R01CA100866-02 (2004): $249375
1R01CA100866-01 (2003): $292275
CDK/histone Deacetylase Inhibition In Acute Leukemia
Steven Grant, Professor
Virginia Commonwealth University Po Box 980568 Richmond, Va 232980568
Grant 5R21CA115260-02 from National Cancer Institute IRG: CONC
Abstract: The goal of this application is to conduct a Phase I trial and correlative laboratory studies in which the cyclin-dependent kinase (CDK) inhibitor flavopiridol (NSC 649890) is combined with the histone deacetylase inhibitor (HDACI) SAHA (NSC 701852) for the treatment of refractory acute leukemia. This concept is based on preclinical results from this and other laboratories indicating that flavopiridol, despite inducing cell cycle arrest, does not promote HDACI-mediated maturation in leukemic cells; instead, it results in a pronounced increase in mitochondrial injury and apoptosis. This phenomenon may stem from flavopiridol-mediated inhibition of the PTEF-b transcription complex and other kinases, leading to down-regulation/inactivation of various anti-apoptotic proteins, including p21CIP1, XIAP, Mcl-1, NF-kappa B, and diminished phosphorylation of RNA pol II carboxy-terminal domain (CTD). Furthermore, both flavopiridol and SAHA have shown promising evidence of activity in acute leukemia in early Phase I trials. These considerations have prompted the development of a CTEP-approved Phase I trial (NCI #P6637) of SAHA administered po TID x 14 d in conjunction with flavopiridol administered as a 1-hr infusion d 1-5 for patients with refractory leukemia. The goals of this application are 1) to establish the maximally tolerated dose (MTD) of SAHA and Flavopiridol administered in this manner; 2) define the recommended Phase II dose (RPTD) for this regimen; 3) characterize the dose-limiting toxicities (DLTs) of these agents; 4) obtain preliminary evidence of activity of the regimen; 5) characterize the pharmacokinetics of flavopiridol and SAHA. Correlative laboratory studies will also be carried out to test the hypotheses that 1) in vivo administration of flavopiridol and SAHA will result in down-regulation/inactivation of p21CIP1, XIAP, Mcl-1, NF-kappa B, and diminished phosphorylation of CTD RNA pol II in primary patient-derived blasts, as observed in preclinical studies; 2) similar events will occur in blasts exposed to these agents ex vivo; and 3) plasma flavopiridol levels will be achieved sufficient to block SAHA-mediated p21CIP1 induction in cultured leukemia cell lines. Information derived from this trial will lay the groundwork for subsequent rationally designed Phase II trials employing a novel strategy involving simultaneous CDK and HDAC inhibition in the treatment of refractory acute leukemia.
Keywords: acute leukemia, amidohydrolase, cyclin dependent kinase, drug screening /evaluation, enzyme inhibitor, flavopiridol, histone, kinase inhibitor, neoplasm /cancer chemotherapy, clinical trial phase I, dosage, human therapy evaluation, nuclear factor kappa beta, pharmacokinetics, phosphorylation, cell line, clinical research, human subject, patient oriented research
Project start date: 2005-08-11
Project end date: 2008-07-31
5R21CA115260-02 (2006): $251828
1R21CA115260-01 (2005): $277243
PROTEASOME/HDAC INHIBITION IN LEUKEMIA/MDS; PHASE I TRIAL AND CORRELATIVE STUDIES
Steven Grant
Virginia Commonwealth University, Po Box 980568, Richmond, Va 23298-0568
Grant 5RC2CA148431-02 from National Cancer Institute
Abstract: The central goal of this GO RC2 application is to leverage the collective resources of three institutions (VCU/Massey Cancer Center, MD Anderson Cancer Center, H. Lee Moffitt Cancer Center) and various associated support mechanisms (i.e., R01, P01, N01, and SPORE) to conduct a mechanism-based Phase I trial of the histone deacetylase inhibitor (HDACI) belinostat (PXD-101) and the proteasome inhibitor (PI) bortezomib in patients with refractory AML, high-risk MDS, CML-blast crisis, and ALL. The second goal is to perform correlative laboratory studies to test the adequacy of methods for monitoring candidate surrogate markers that may predict for disease responsiveness and help to define mechanisms of resistance to this regimen in future efficacy-based trials (e.g., Phase II). Previous studies from our laboratories documented pronounced synergism between HDACIs and PIs in malignant hematopoietic cells, including human leukemia cells. Mechanisms responsible for synergistic interactions are likely to be multi-factorial, including PI-mediated inhibition of HDACI-induced NF-B activation, down-regulation of NF-B-dependent anti-apoptotic proteins (Bcl-xL, XIAP), HDACI-mediated up-regulation of Bim, and disruption of aggresome function. In addition, evidence suggests that HDACIs disrupt proteasome function, raising the possibility that combined treatment with these agents may result in enhanced proteasome inhibition. Notably, recent preclinical evidence from our laboratories indicates that very low (e.g., nM) concentrations of belinostat and bortezomib interact in a highly synergistic manner in cultured and primary AML blasts to induce apoptosis in association with diminished nuclear p65/RelA localization, down-regulation of NF-B-dependent proteins (Bcl-xL and XIAP), and up- regulation of Bim. Despite this preclinical evidence, a strategy combining HDACIs with PIs has not yet been evaluated in AML, MDS, and related acute leukemias. Specific Aim #1 of this proposal is to conduct a Phase I trial of belinostat given IVP days 1-5 and 8-12 of a 3-wk schedule in conjunction with bortezomib given IVP twice weekly x two weeks and to identify the RPTD (recommended Phase II doses) for future Phase II trials. Secondary aims are to identify the dose-limiting toxicities of this regimen, and to gain preliminary insights into the potential therapeutic efficacy of this strategy. Specific Aim #2 of this proposal is to test the adequacy of methods for monitoring candidate correlative pharmacodynamic determinants in leukemic blast cells prior to and 24 hr after treatment with belinostat/bortezomib, focusing on events observed in vitro in preclinical studies e.g., diminished p65/RelA nuclear localization by digitized fluorescence microscopy; Bcl-xL/XIAP down- regulation and Bim up-regulation by Western blot analysis; and inhibition of 20S proteasome activity. The successful conduct of this trial and performance of correlative laboratory studies could serve as a prototype for future partnerships between the NCI, academia, and the pharmaceutical industry in the development of novel, mechanism-based anti-cancer therapeutic strategies involving two or more investigational agents. Acute myelogenous leukemia and related diseases (myelodysplasic syndrome, acute lymphocytic leukemia, chronic myelogenous leukemia in blast crisis) are responsible for significant morbidity and mortality. If successful, the current proposal could lead to the development of a new and potentially more effective treatment strategy for these diseases, and could also help to identify laboratory correlates that might predict for disease responsiveness in individual patients
Keywords: 20S Catalytic Proteasome; 20S Core Proteasome; 20S Proteasome; 20S Proteosome; ALL - Acute Lymphocytic Leukemia; AML - Acute Myeloid Leukemia; API3; Academia; Accounting; Acetylation; Acute Lymphoid Leukemia; Acute leukemia; Address; After Care; After-Treatment; Aftercare; Apoptosis; Apoptosis Pathway; Apoptotic; Assay; BIRC4; BIRC4 gene; Bioassay; Biochemical; Biologic Assays; Biological Assay; Blast Cell; Blast Crisis; Blast Phase; Blast Phase CML; Blast Phase Chronic Granulocytic Leukemia; Blast Phase Chronic Myelocytic Leukemia; Blast Phase Chronic Myelogenous Leukemia; Blast Phase Chronic Myeloid Leukemia; Blastic Phase CML; Blastic Phase Chronic Granulocytic Leukemia; Blastic Phase Chronic Myelocytic Leukemia; Blastic Phase Chronic Myelogenous Leukemia; Blastic Phase Chronic Myeloid Leukemia; Blasts; Blood (Leukemia); Blotting, Western; Bortezomib; Cancer Center; Cell Death; Cell Death, Programmed; Cells; Chronic Myeloid Leukemia; Clinical; Clinical Trials, Phase I; Clinical Trials, Phase II; Combined Modality Therapy; Correlative Study; Development; Disease; Disorder; Dose; Dose-Limiting; Down-Regulation; Down-Regulation (Physiology); Downregulation; Drug Industry; Dysfunction; Early-Stage Clinical Trials; Event; Ferrata cell; Fluorescence Microscopy; Functional disorder; Future; Goals; HDAC; HDAC Agent; HDAC Proteins; Hematohistioblast; Hematologic Cancer; Hematologic Malignancies; Hematologic Neoplasms; Hematological Malignancies; Hematological Neoplasms; Hematological Tumor; Hematopoietic; Hematopoietic Cancer; Hemocytoblast; Hemohistioblast; Histone Deacetylase; Histone Deacetylase Inhibitor; Human; Human, General; IAP-like protein, human; ILP; In Vitro; Individual; Indolent; Industry, Pharmaceutic; Institution; Laboratories; Laboratory Study; Lead; Leukemia, Granulocytic, Chronic; Leukemia, Lymphocytic, Acute; Leukemia, Myelocytic, Acute; Leukemias, General; Lymphoblastic Leukemia, Acute; MIHA; Macropain; Macroxyproteinase; Malignant; Malignant - descriptor; Malignant Hematologic Neoplasm; Malignant lymphoid neoplasm; Man (Taxonomy); Man, Modern; Mediating; Methods; Microscopy, Fluorescence; Microscopy, Light, Fluorescence; Monitor; Morbidity; Morbidity - disease rate; Mortality; Mortality Vital Statistics; Multicatalytic Proteinase; Multimodal Therapy; Multimodal Treatment; Multimodality Treatment; Multiple Myeloma; Myeloblastic Leukemia, Acute; Myelocytic Leukemia, Chronic; Myelogenous Leukemia, Acute; Myelogenous Leukemia, Chronic; Myeloid Leukemia, Chronic; Myeloma, Plasma-Cell; Nuclear; Patients; Pb element; Performance; Pharmaceutical Industry; Pharmacodynamics; Phase; Phase 1 Clinical Trials; Phase 2 Clinical Trials; Phase I Clinical Trials; Phase I Study; Phase II Clinical Trials; Physiopathology; Precursor Cell Lymphoblastic Leukemia; Precursor Lymphoblastic Leukemia; Prosome; Proteasome; Proteasome Endopeptidase Complex; Proteasome Inhibition; Proteasome Inhibitor; Proteins; Proteosome; Refractory; Regimen; Reproduction spores; Research Resources; Resources; SCHED; Safety; Schedule; Spores; Surrogate Markers; Syndrome; Testing; Therapeutic; Toxic effect; Toxicities; Treatment Efficacy; Up-Regulation; Up-Regulation (Physiology); Upregulation; Western Blotting; Western Blottings; Western Immunoblotting; X-linked IAP; XIAP; XIAP protein; acute granulocytic leukemia; acute lymphatic leukemia; acute lymphogenous leukemia; acute lymphomatic leukemia; acute myeloid leukemia; acute nonlymphocytic leukemia; anti-cancer therapeutic; anticancer therapeutic; base; cell transformation; cell type; combination therapy; combined modality treatment; combined treatment; disease/disorder; effective therapy; gene product; hILP protein; heavy metal Pb; heavy metal lead; high risk; in vivo; insight; leukemia; lymphoid malignancy; multicatalytic endopeptidase complex; multimodality therapy; myeloma; myelomatosis; necrocytosis; novel; p65; pathophysiology; phase 1 study; phase 1 trial; phase 2 study; phase 2 trial; phase I trial; phase II trial; pre-clinical; preclinical; preclinical study; pro-apoptotic protein; protein blotting; protocol, phase I; protocol, phase II; prototype; public health relevance; resistance mechanism; resistant mechanism; response; study, phase II; synergism; therapeutic efficacy; therapeutically effective; transformed cells; treatment strategy; x-linked IAP protein
Relevance: Acute myelogenous leukemia and related diseases (myelodysplasic syndrome, acute lymphocytic leukemia, chronic myelogenous leukemia in blast crisis) are responsible for significant morbidity and mortality. If successful, the current proposal could lead to the development of a new and potentially more effective treatment strategy for these diseases, and could also help to identify laboratory correlates that might predict for disease responsiveness in individual patients
Project start date: 2009-09-30
Project end date: 2011-08-31
Budget start date: 1-SEP-2010
Budget end date: 31-AUG-2011
PFA/PA: RFA-OD-09-004
5RC2CA148431-02 (2010): $589802
1RC2CA148431-01 (2009): $591990
Flavopiridol And Imatinib In Bcr/Abl+ Leukemia
Steven Grant, Professor
Virginia Commonwealth University Po Box 980568 Richmond, Va 232980568
Grant 5R21CA106139-02 from National Cancer Institute IRG: CONC
Abstract: Recent evidence suggests that neoplastic cells are particularly susceptible to a strategy involving simultaneous interruption of survival-associated signal transduction and cell cycle regulatory pathways. Consistent with this notion, we have observed in preclinical studies that the cyclin-dependent kinase inhibitor flavopiridol (NSC 649890) interacts synergistically with the Bcr/Abl kinase inhibitor imatinib (STI571; Gleevec) to induce mitochondrial injury, caspase activation and apoptosis in Bcr/Abl+ human leukemia cells, including those highly resistant to imatinib. These events are associated with multiple perturbations in survival signaling and cell cycle-related pathways, including down-regulation of Mcl-1 and Bcl-xL, reduced expression of cyclin D1, activation of JNK, and inactivation of CREB and Stat5. Based upon these findings, a multi-institutional Phase I trial has been developed in which patients with progressive CML (chronic and accelerated phase) or CML-BC or Philadelphia chromosome+ AML or ALL will be treated with escalating doses of daily imatinib in conjunction with flavopiridol administered as a 1-hr infusion weekly x 3 q month. The goals of this Phase I trial are to define the MTD for these agents, characterize dose-limiting toxicities, and gain preliminary information regarding activity of the regimen. Correlative laboratory studies will test the hypothesis that in vivo administration of imatinib in conjunction with flavopiridol will induce perturbations in apoptotic regulatory proteins in peripheral blood Bcr/Abl+ cells (e.g., diminished expression of Mcl-1, Bcl-xL, and cyclin D1, inactivation of Stat5 and CREB, activation of JNK) similar to those observed in Bcr/Abl+ cell lines exposed to these agents in vitro. Other studies will investigate a) effects of the imatinib/flavopiridol regimen on Stat5 phosphorylation of Bcr/Abl+ peripheral blood cells by flow cytometry; b) the pharmacokinetics of imatinib and flavopiridol when administered together; and c) the presence of Bcr/Abl mutations as well as increased Bcr/Abl expression/activity in cells from imatinib-resistant patients, and their possible relationship to imatinib/flavopiridol pharmacodynamics. Information derived from this trial will provide a foundation for a successor Phase II trial and correlative laboratory studies which will address issues of regimen activity and imatinib/flavopiridol molecular interactions more definitively.
Keywords: acute lymphocytic leukemia, acute myelogenous leukemia, antineoplastic, chronic myelogenous leukemia, combination chemotherapy, enzyme inhibitor, flavopiridol, human therapy evaluation, neoplasm /cancer chemotherapy, JUN kinase, biological signal transduction, cell cycle, clinical trial phase I, dosage, monocyte, neoplasm /cancer pharmacology, oncoprotein, pharmacokinetics, transcription factor, clinical research, flow cytometry, human subject, patient oriented research
Project start date: 2003-09-11
Project end date: 2006-08-31
5R21CA106139-02 (2004): $271299
1R21CA106139-01 (2003): $297053
PHASE I TRIAL OF BORTEZOMIB AND ROMIDEPSIN IN CLL AND SMALL CELL LYMPHOMA
Steven Grant
Virginia Commonwealth University, Po Box 980568, Richmond, Va 23298-0568
Grant 5R21CA137823-02 from National Cancer Institute
Abstract: Previous studies from this and other laboratories have established that histone deacetylase inhibitors (HDACIs) and proteasome inhibitors such as bortezomib interact synergistically to induce apoptosis in malignant human hematopoietic cells. In chronic lymphocytic leukemia (CLL) cells, postulated mechanisms of synergism have focused on bortezomib-mediated blockade of HDACI-induced RelA acetylation and activation of the canonical and alternative NF-B pathways, resulting in down regulation of NF-B-dependent survival proteins (e.g., Bcl-xL and XIAP). Very recently, we have observed that when co-administered in vitro at extremely low concentrations (i.e. 3-5 nM each), the Class I HDACI romidepsin (depsipeptide; FK228) interacts with bortezomib to induce very pronounced apoptosis in fresh primary CLL cells as well as .CLL cell lines. Furthermore, these events are associated with prevention of romidepsin-induced activation of the classical and alternative NF-B pathways, down regulation of the NF-B dependent proteins Bcl-xL and XIAP, and induction of the pro-apoptotic protein Bim. We now propose to begin testing the in vivo implications of these preclinical findings by conducting a Phase I trial. The specific aims of this proposal are First, to determine the maximum tolerated dose (MTD) for the combination of bortezomib and romidepsin administered weekly x 3 every 4 weeks in patients with chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL); to determine the safety and describe the toxicities of the combination; and to document activity of the combination observed in the course of the dose finding study. Second, to demonstrate adequate techniques for the assessment of pharmacodynamic responses of CLL cells to the combination with respect to effects on activation of the canonical and alternative NF-B pathways (nuclear RelA and p52 as a marker of p100 processing), expression of the NF-B-dependent proteins XIAP and Bcl-xL, and expression of the pro-apoptotic protein Bim; and to document pharmacodynamic responses observed in the course of the dose finding study. Studies from our laboratory have shown a potent interaction between the histone deacetylase inhibitor romidepsin and the proteosome inhibitor in inducing cell death in primary chronic lymphocytic leukemia (CLL) cells. The purpose of this study is to determine the maximum tolerated dose (MTD) for the combination administered weekly x 3 every 4 weeks in patients with chronic lymphocytic leukemia/small cell lymphocytic lymphoma (CLL/SLL), to determine the safety and describe the toxicities of the combination, and to document activity of the combination observed in the course of the dose finding study. Further, the purpose is to demonstrate adequate techniques for the assessment of pharmacodynamic responses of CLL cells to the combination with respect to effects on activation of the canonical and alternative NF-B pathways, expression of selected NF-B-dependent proteins, and expression of pro-apoptotic protein Bim, and to document pharmacodynamic responses observed in the course of the dose finding study. This will position us to perform future trials that will determine the effectiveness of this novel drug combination in patients with CLL or SLL and address the validity of our preclinical pharmacodynamic observations
Keywords: 20S Catalytic Proteasome; 20S Core Proteasome; 20S Proteasome; 20S Proteosome; API3; Acetylation; Address; Apoptosis; Apoptosis Pathway; B-Cell CLL; B-Cell Chronic Lymphocytic Leukemia; B-Cell Chronic Lymphogenous Leukemia; B-Cell Chronic Lymphoid Leukemia; B-Lymphocytic Leukemia; B-Lymphocytic Leukemia, Chronic; BIRC4; BIRC4 gene; Blood (Leukemia); Bortezomib; Cell Death; Cell Death, Programmed; Cell Line; Cell Lines, Strains; CellLine; Cells; Chronic; Chronic B-Cell Leukemias; Chronic Lymphatic Leukemia; Chronic Lymphocytic Leukemia; Chronic Lymphogenous Leukemia; Clinical Trials, Phase I; Depsipeptide Antibiotic; Depsipeptides; Dose; Down-Regulation; Down-Regulation (Physiology); Downregulation; Drug Combinations; Early-Stage Clinical Trials; Effectiveness; Event; Future; HDAC Agent; Hematopoietic; Histone Deacetylase Inhibitor; Human; Human, General; IAP-like protein, human; ILP; In Vitro; Laboratories; Leukemia, B-Cell; Leukemias, General; Lymphoblastic Leukemia, Chronic; Lymphocyte; Lymphocytic; Lymphocytic Leukemia, B-Cell; Lymphocytic Leukemia, Chronic, B-Cell; Lymphoma, Lymphocytic; Lymphoma, Lymphocytic, Diffuse, Well-Differentiated; Lymphoma, Lymphocytic, Plasmacytoid; Lymphoma, Lymphocytic, Well-Differentiated; Lymphoma, Small Lymphocytic; Lymphoma, Small Lymphocytic, Plasmacytoid; Lymphoma, Small-Cell; Lymphoplasmacytoid Lymphoma, CLL; MIHA; Macropain; Macroxyproteinase; Malignant; Malignant - descriptor; Man (Taxonomy); Man, Modern; Maximal Tolerated Dose; Maximally Tolerated Dose; Maximum Tolerated Dose; Mediating; Methods and Techniques; Methods, Other; Multicatalytic Proteinase; Nuclear; Pathway interactions; Patients; Pharmacodynamics; Phase 1 Clinical Trials; Phase I Clinical Trials; Phase I Study; Position; Positioning Attribute; Prevention; Process; Prosome; Proteasome; Proteasome Endopeptidase Complex; Proteasome Inhibitor; Proteins; Proteosome; Safety; Small B-Cell Lymphocytic Lymphoma; Small Cell Lymphoma; Small-Cell Lymphoma; Techniques; Testing; Toxic effect; Toxicities; X-linked IAP; XIAP; XIAP protein; chronic lymphoid leukemia; cultured cell line; gene product; hILP protein; in vivo; inhibitor; inhibitor/antagonist; leukemia; lymph cell; multicatalytic endopeptidase complex; necrocytosis; novel; pathway; phase 1 study; phase 1 trial; phase I trial; pre-clinical; preclinical; pro-apoptotic protein; protein expression; protocol, phase I; response; synergism; x-linked IAP protein
Relevance: Studies from our laboratory have shown a potent interaction between the histone deacetylase inhibitor romidepsin and the proteosome inhibitor in inducing cell death in primary chronic lymphocytic leukemia (CLL) cells. The purpose of this study is to determine the maximum tolerated dose (MTD) for the combination administered weekly x 3 every 4 weeks in patients with chronic lymphocytic leukemia/small cell lymphocytic lymphoma (CLL/SLL), to determine the safety and describe the toxicities of the combination, and to document activity of the combination observed in the course of the dose finding study. Further, the purpose is to demonstrate adequate techniques for the assessment of pharmacodynamic responses of CLL cells to the combination with respect to effects on activation of the canonical and alternative NF-¿B pathways, expression of selected NF-:B-dependent proteins, and expression of pro-apoptotic protein Bim, and to document pharmacodynamic responses observed in the course of the dose finding study. This will position us to perform future trials that will determine the effectiveness of this novel drug combination in patients with CLL or SLL and address the validity of our preclinical pharmacodynamic observations
Project start date: 2009-08-06
Project end date: 2011-07-31
Budget start date: 27-AUG-2010
Budget end date: 31-JUL-2011
PFA/PA: PAR-08-025
5R21CA137823-02 (2010): $262278
Sponsored Links Excellgen http://Excellgen.com
1R21CA137823-01A1 (2009): $274653
Phase II And Correlative Studies Of Bryo/HiDAC
Steven Grant, Professor
Internal Medicinevirginia Commonwealth University
po Box 980568
richmond, Va 232980568
Grant 1R21CA092950-01 from National Cancer Institute IRG: ZRG1
Abstract: Preclinical studies have demonstrated that the macrocyclic lactone bryostatin 1 (NSC 399555), which initially activates and subsequently down-regulates the serine/threonine kinase protein kinase C, sensitizes human leukemia cells to 1 -B-D-arabinofuranosylcytosine (ara-C)-induced apoptosis in a dose- and schedule-dependent manner, resulting in synergistic antileukemic effects for the combination. Based upon these and other findings, a multi-institutional Phase I trial in which escalating doses of bryostatin 1 were given as a 24-hr continuous infusion before and after 4 doses of high-dose-ara-C (HiDAC; 1.5 gm/sq.m. q 12h x 4 on days 2-4 and 9-10; 8 total doses) in patients with refractory/relapsed acute leukemia has recently been completed and has identified the bryostatin 1 MTD as 50 ug/sq.m. Initial results of this Phase I trial were encouraging, with 5 objective CRs and one 6+ month leukemia-free interval obtained in a heavily pretreated patient population. The goal of the present application is to conduct a formal multi-institutional Phase II trial of bryostatin 1 and HiDAC in patients with refractory or relapsed acute leukemia, CML-myeloid blast crisis, or untreated high-risk acute leukemia to define the toxicities and antileukemic activity of this regimen more rigorously. Correlative laboratory studies will be conducted in parallel to determine whether in vivo administration of bryostatin 1 (a) down-regulates leukemic blast PKC activity; (b) modifies ex vivo expression of p21 CIP 1, Bcl-2, or phospho-MAPK in ara-C-treated blasts; (c) sensitizes blasts to ara-C-induced mitochondrial damage and apoptosis ex vivo; or (d) increases ara-CTP formation. Additional studies will establish whether sequential ex vivo exposure of blasts to ara-C followed by bryostatin 1 maximizes apoptosis in the subset of leukemic specimens susceptible to bryostatin 1-induced maturation. Finally, attempts will be made to determine which if any of these laboratory determinants correlates with clinical responsiveness to the HiDAC/bryostatin 1 regimen. Information derived from this trial will aid in the design of successor studies in which bryostatin 1 or other novel agents acting through signal transduction pathways are combined with established cytotoxic agents in the treatment of leukemia and possibly other hematologic malignancies
Keywords: acute leukemia, bryostatin, cytosine arabinoside, human therapy evaluation, neoplasm /cancer chemotherapy BCL2 gene /protein, apoptosis, clinical trial phase II /III /IV, combination chemotherapy, drug metabolism, enzyme activity, enzyme induction /repression, mitochondria, mitogen activated protein kinase, neoplasm /cancer relapse /recurrence, oncoprotein p21, protein kinase C cryopreservation, human subject, patient oriented research
Project start date: 2001-07-16
Project end date: 2003-06-30
1R21CA092950-01 (2001): $323820
PHASE IB STUDY OF BRYOSTATIN 1--NSC339555
Steven Grant, Professor
Institution:
Grant 5M01RR000065-340457 from National Center For Research Resources
Abstract: NO AVAILABLE
Keywords: bryostatin, human therapy evaluation, clinical trial phase I, clinical research, human subject
NF-KAPPAB INHIBITORS AND DIFFERENTIATION-INDUCERS IN LEUKEMIA
Steven Grant, Professor
Virginia Commonwealth University, Po Box 980568, Richmond, Va 23298-0568
Grant 5R01CA093738-08 from National Cancer Institute
Abstract: In this renewal application, we propose to extend our efforts to develop novel antileukemic strategies based on the concept of disrupting/modulating the actions of differentiation-inducing agents, primarily histone deacetylase inhibitors (HDACIs), to promote mitochondrial injury and apoptosis in leukemia cells. In the preceding application, we demonstrated that the CDK inhibitor flavopiridol triggered multiple perturbations in leukemia cells, including down-regulation of p21CIP1, XIAP, Mcl-1, and NF-kappaB, that collectively blocked HDACI-mediated maturation and reciprocally potentiated apoptosis, resulting in pronounced antileukemic synergism. These studies resulted in several multi-institutional Phase I trials in patients with hematologic and non-hematologic malignancies. Insights generated during the preceding funding period have led us to the hypothesis that NF-kappaB/p65 acetylation/nuclear translocation/activation plays a pivotal role in protecting leukemic cells from HDACI-mediated lethality, and that interference with these events at multiple levels (e.g., through inhibition of IKKs, particularly IKKbeta, proteasome inhibition, or activation of the SIRT1 HDAC by resveratrol) potently trigger leukemic cell apoptosis through the selective induction of oxidative injury. The specific aims of this proposal are a) to elucidate, through both pharmacologic and genetic strategies, the functional roles of disruption of p65 acetylation/phosphorylation, nuclear translocation, and activation in synergistic antileukemic interactions between these agents; b) to determine whether and to what extent downregulation/inactivation of NF-kappaB antioxidant target genes, particularly MnSOD, Trx/TrxR, and GPx1, culminate in oxidative injury and contribute to antileukemic activity and selectivity; c) to test the hypothesis that sustained activation of the stress-related JNK pathway plays a critical role in mediating the antileukemic activity of these regimens; and d) to perform parallel studies in primary leukemic and leukemia stem cells (LSC), and to employ xenograft and NOD/SCID murine models, to establish a basis for the selectivity of this strategy. The relevance of this proposal is that the information generated may provide a rational foundation for the development of entirely novel antileukemic regimens combining inhibitors of an important survival pathway (NF-kappaB) and a promising new class of antileukemic agents (HDAC inhibitors) in the treatment of acute leukemia and potentially other hematologic malignancies in humans
Keywords: (-)cis-5, 7-dihydroxy-2-(2-chlorophenyl)-8-(4-(3-hydroxy-1-methyl)piperidinyl)-4H-1-benzopyran-4-one; 2-(1-ethyl-2-hydroxyethylamino)-6-benzylamino-9-isopropylpurine; 3, 4`, 5-stilbenetriol; 3, 5, 4`-trihydroxystilbene; 4H-1-Benzopyran-4-one, 2-(2-chlorophenyl)-5, 7-dihydroxy-8-(3-hydroxy-1-methyl-4-piperidinyl)-, hydrochloride, (-)-cis-; API3; Acetylation; Acute leukemia; Agonist; Animal Model; Animal Models and Related Studies; Antioxidants; Apoptosis; Apoptosis Pathway; BIRC4; BIRC4 gene; Biological Models; Blast Cell; Blasts; Blood (Leukemia); Bortezomib; CDK Inhibitor Protein; CDKI Protein; CYC 202; CYC202; Cell Communication and Signaling; Cell Cycle; Cell Death, Programmed; Cell Division Cycle; Cell Maturation; Cell Signaling; Cells; Clinical Trials, Phase I; Complex; Cyclin Kinase Inhibitor; Cyclin-Dependent Kinase Inhibitor; Development; Differentiating Agents; Differentiation Agents; Differentiation Inducer; Down-Regulation; Down-Regulation (Physiology); Downregulation; Early-Stage Clinical Trials; Enzymes; Event; Exhibits; Ferrata cell; Flavopirodol/HMR-1275; Foundations; Funding; Gene Targeting; Gene Transcription; Genetic; Genetic Transcription; Glutathione[{..}]hydrogen-peroxide oxidoreductase; Goals; HD3; HDAC; HDAC Agent; HDAC Proteins; HDAC3; HDAC3 gene; Hematohistioblast; Hematologic Cancer; Hematologic Malignancies; Hematologic Neoplasms; Hematological Malignancies; Hematological Neoplasms; Hematological Tumor; Hematopoietic Cancer; Hemocytoblast; Hemohistioblast; Heterograft; Histone Deacetylase; Histone Deacetylase Inhibitor; Human; Human, General; ILP; Immunoglobulin Enhancer-Binding Protein; Injury; Intracellular Communication and Signaling; JN Kinase; JNK; JNK Mitogen-Activated Protein Kinases; JNK1; JNK1 Kinase; JNK1 protein; JNK1A2; JNK21B1/2; Killings; LEUKCL; Leukemias, General; Leukemic Cell; MAP Kinase 8; MAP Kinase 8 Gene; MAPK8; MAPK8 Mitogen-Activated Protein Kinase; MAPK8 gene; MIHA; Malignant Hematologic Neoplasm; Mammals, Mice; Man (Taxonomy); Man, Modern; Mediating; Mice; Mitochondria; Mitogen-Activated Protein Kinase 8; Model System; Modeling; Models, Biologic; Mother Cells; Murine; Mus; NF-kB; NF-kappa B; NF-kappaB; NFKB; Non-Hematologic Cancer; Non-Hematologic Malignancy; Nuclear; Nuclear Factor kappa B; Nuclear Transcription Factor NF-kB; Nuclear Translocation; PRKM8; Pathway interactions; Patients; Phase 1 Clinical Trials; Phase I Clinical Trials; Phase I Study; Phosphorylation; Play; Progenitor Cells; Proteasome Inhibition; Proteasome Inhibitor; Protein Phosphorylation; RNA Expression; RPD3; RPD3-2; Refractory; Regimen; Reporting; Resveratrol; Role; SAHA; SAP Kinase-1; SAPK/JNK; SAPK1; SAPK1 Mitogen-Activated Protein Kinase; SAPK1/JNK; Signal Pathway; Signal Transduction; Signal Transduction Systems; Signaling; Solid Neoplasm; Solid Tumor; Stem Cell Leukemia; Stem cells; Stress; Stress-Activated Protein Kinase JNK1; Stress-Activated Protein Kinase gamma; Suberoylanilide Hydroxamic Acid; Targetings, Gene; Testing; Thioredoxin; Transcription; Transcription Factor NF-kB; Transcription, Genetic; Transplantation, Heterologous; Vorinostat; XIAP; Xenograft; Xenograft procedure; Xenotransplantation; analog; anti-oxidant; antileukemic agent; base; biological signal transduction; c-jun Amino-Terminal Kinase; c-jun Kinase-1; c-jun N-Terminal Kinase; c-jun N-Terminal Kinase 1; clinical relevance; clinically relevant; cyclin T; cyclin T1; flavopiridol; glutathione peroxidase; in vivo; inhibitor; inhibitor/antagonist; insight; jun-NH2-Terminal Kinase; kappa B Enhancer Binding Protein; leukemia; mitochondrial; model organism; nonhematologic cancer; novel; nuclear factor kappa beta; p65; pathway; phase 1 study; phase 1 trial; phase I trial; protocol, phase I; roscovitine; social role; stress-activated protein kinase 1; suberanilohydroxamic acid; synergism; thioredoxin reductase
Project start date: 2002-01-01
Project end date: 2012-07-31
Budget start date: 1-AUG-2010
Budget end date: 31-JUL-2011
5R01CA093738-08 (2010): $253047
3R01CA093738-08S1 (2010): $57444
5R01CA093738-07 (2009): $253047
3R01CA093738-07S1 (2009): $55731
5R01CA093738-06 (2008): $253047
2R01CA093738-05A2 (2007): $253047
PHASE 1 TRIAL--BRYOSTATIN 1 AND HIGH DOSE ARA C (HIDAC)
Steven Grant, Professor
Virginia Commonwealth University Po Box 980568 Richmond, Va 232980568
Grant 1R03CA077141-01 from National Cancer Institute IRG: NCI
Abstract: Bryostatin 1 is a macrocyclic lactone protein kinase C (PKC) activator that has recently undergone phase I evaluation in humans. Preclinical evidence indicates that bryostatin 1 increases the susceptibility of human leukemic cells to 1-beta-D-arabinofuranosylcytosine (ara-C)- induced apoptosis in a dose- and schedule-dependent manner, and that this phenomenon is associated with bryostatin 1-mediated PKC down- regulation. In vivo administration of bryostatin 1 has been shown to down-regulate splenocyte PKC activity in a murine model. Moreover, in a recently completed phase Ib pharmacodynamic trial in humans, a fixed dose of bryostatin 1 (25 g/M2) administered according to three schedules was observed to induce down-regulation of peripheral blood mononuclear cell PKC activity in a subset of patients. Based upon these findings, a phase I trial has been designed and subsequently approved by CTEP in which escalating doses of bryostatin 1 will be administered before and after high-dose ara-C (HIDAC) in patients with refractory leukemia. The goals of this trial are (1) to identify the MTD of bryostatin 1 administered as a 24-hour infusion immediately before and after 4 courses of HIDAC (1.5 gM/M2 q hx4 on days 1+2 and 9+10); (2) to identify the dose-limiting toxicities of this regimen; (3) to assess the effects of bryostatin 1 on HIDAC pharmacokinetics, and (4) to determine, using a platelet aggregation-based bioassay, whether escalation of the bryostatin 1 dose will lead to detectable plasma levels. Funds are now requested for clinical and data management support for this clinical trial, as well as correlative pharmacokinetic studies. It is anticipated that this trial could serve as a prototype for future studies in which agents acting through signal transduction pathways are combined with conventional cytotoxic agents in the treatment of patients with refractory hematopoietic and other malignancies.
Keywords: antileukemic agent, blood disorder chemotherapy, bryostatin, chronic myelogenous leukemia, combination chemotherapy, cytosine arabinoside, human therapy evaluation, neoplasm /cancer chemotherapy, pharmacokinetics, blood /lymphatic pharmacology, clinical trial phase I, dosage, drug administration rate /duration, drug adverse effect, drug interaction, platelet aggregation, bioassay, blood chemistry, clinical research, human subject, injection /infusion, plasma
Project start date: 1997-09-30
Project end date: 1999-09-29
1R03CA077141-01 (1997): $72500
Sponsored Links Excellgen http://Excellgen.com
MODULATION OF ARA-C INDUCED APOPTOSIS BY BRYOSTATIN 1
Steven Grant, Professor
Virginia Commonwealth University Po Box 980568 Richmond, Va 232980568
Grant 5R01CA063753-09 from National Cancer Institute IRG: ET
Abstract: Applicant s ) The goal of this renewal application is to elucidate further the basis by which agents that down-regulate (bryostatin 1) or inhibit (UCN-01, safingol) protein kinase C (PKC) promote leukemic cell apoptosis by ara-C and other nucleoside analogs. Evidence generated during the preceding period of support suggests that interruption of the PKC signal transduction pathway may promote cell death by three distinct mechanisms (a) dysregulation of cell cycle-related events, particularly induction of cyclin-dependent kinase inhibitors (CDKIs); (b) redirection of signals away from cytoprotective survival (e.g., MAPK/ERK) toward stress-related (e.g., SAPK/JNK) pathways; and (c) phosphorylation of the Bcl-2 protein, promoting mitochondrial permeability transition and circumvention of the block to capase activation. To examine these possibilities, (1) Human leukemic cells (HL-60, U937) stably overexpressing Bcl-2, Bcl-xL, and phosphorylation loop deletant mutants will be employed to determine whether phosphorylation of anti-apoptotic proteins is responsible for potentiation of ara-C mediated apoptosis by PKC inhibition/down-regulation, (2) Analogously, HL-60 and U937 cells expressing antisense p21WAF1+/-p27KIP1 will be used to characterize the effect of CDKI dysregulation on ara-C mediated apoptosis and its potentiation by PKC inhibitors/down-regulators, (3) The effects of enforced expression of p53 will be examined with respect to leukemic cell differentiation and modulation of ara-C induced cell death by bryostatin 1/PKC inhibitors, (4) Direct evidence for the participation of stress and survival pathways in cell death decisions will be obtained through the use of SEK1/JNK1 dominant-negative and ER-inducible Raf/MAPK mutants, (5) Findings will be extended to include another clinically important nucleoside analog, 2,2(1)-difluorodeoxcytidine (gemcitabine) and (6) Finally, the ability of pharmacologically relevant concentrations of bryostatin 1, UCN-01, and safingol to potentiate ara-C- (and gemcitabine)-mediated apoptosis in primary leukemic myeloblasts ex vivo will be explored. Information derived from these studies will lay the foundation for a novel approach to leukemia therapy aimed at enhancing the activity of effective antileukemic drugs via combination with agents that interrupt the PKC signal transduction pathway.
Keywords: antileukemic agent, apoptosis, biological signal transduction, bryostatin, cytosine arabinoside, enzyme activity, myelogenous leukemia, protein kinase C, DNA damage, antisense nucleic acid, cell differentiation, cyclin dependent kinase, cytotoxicity, enzyme inhibitor, fludarabine, gene expression, mitogen activated protein kinase, mutant, myeloid stem cell, oncoprotein p21, pharmacokinetics, phosphorylation, staurosporine, tumor suppressor gene, tissue /cell culture
Project start date: 1994-06-01
Project end date: 2004-04-30
5R01CA063753-09 (2002): $314725
5R01CA063753-08 (2001): $272468
5R01CA063753-07 (2000): $1
5R01CA063753-06 (1999): $1
PHASE I/LABORATORY STUDIES BRYOSTATIN/FLVDARA IN CLL/NHL
Steven Grant, Professor
Internal Medicinevirginia Commonwealth University
po Box 980568
richmond, Va 232980568
Grant 5R21CA087056-02 from National Cancer Institute IRG: ET
Abstract: Applicant´s ) The purine analog F-ara-AMP (fludarabine) is among the most active agents available against B cell malignancies such as CLL and indolent non-Hodgkin´s lymphoma. The signal transduction modulator bryostatin 1 (NSC 339555), a macrocyclic lactone activator/down-regulator of PKC currently under extensive clinical evaluation, has also displayed activity against these disorders in early Phase I trials. Furthermore, preclinical in vitro and in vivo studies have documented synergistic, sequence-dependent anti-leukemic interactions between these agents, a phenomenon that may involve either (a) a diminished threshold for apoptosis or (b) potentiation of leukemic cell differentiation. Based upon these considerations, a multicenter Phase I trial has been initiated in which patients with progressive CLL or refractory indolent NHL are randomized to receive escalating doses of bryostatin 1 as a 24-hr infusion (initial dose 16 mcg/sq m) either before, or after, a 5-day course of fludarabine (initial dose 12.5 mg/sq m/d). Preliminary results of this study are encouraging, with 2 complete and 3 partial objective responses (out of 12 patients) at the initial Phase I drug dose levels. The specific aims of this application are (1) To determine the MTD for bryostatin 1 when given before or after a standard course of fludarabine; (2) To characterize the dose-limiting and sequence-dependent toxicities of this regimen; (3) To test the hypotheses that in vivo administration of bryostatin 1 can down-regulate total PKC activity in primary CLL lymphocytes, and that this activity correlates with biological actions; (4) To determine (a) whether prior in vivo administration of bryostatin 1 sensitizes CLL lymphocytes to fludarabine-mediated apoptosis ex vivo, and (b) whether subsequent ex vivo exposure to bryostatin 1 promotes apoptosis in CLL cells previously exposed to fludarabine in vivo; (5) To test the hypothesis that in vivo administration of bryostatin 1 and fludarabine can induce differentiation of CLL lymphocytes. Information derived from these studies will assist in the rational design of successor Phase II efficacy trials of bryostatin 1 and fludarabine in B-cell malignancies, provide mechanistic insights into in vivo interactions between these agents and help to identify intermediate laboratory-based markers that are capable of predicting disease responsiveness to this regimen
Keywords: bryostatin, chronic lymphocytic leukemia, combination chemotherapy, fludarabine, human therapy evaluation, neoplasm /cancer chemotherapy, nonHodgkin`s lymphoma clinical trial phase I, drug administration rate /duration, neoplasm /cancer pharmacology clinical research, human subject
Project start date: 2000-05-08
Project end date: 2002-04-30
5R21CA087056-02 (2001): $143362
1R21CA087056-01 (2000): $143425
MODULATION OF ARA-C INDUCED APOPTOSIS BY BRYOSTATIN 1
Steven Grant, Professor
Internal Medicinevirginia Commonwealth University
po Box 980568
richmond, Va 232980568
Grant 3R01CA063753-07S1 from National Cancer Institute IRG: ET
Abstract: Applicant´s ) The goal of this renewal application is to elucidate further the basis by which agents that down-regulate (bryostatin 1) or inhibit (UCN-01, safingol) protein kinase C (PKC) promote leukemic cell apoptosis by ara-C and other nucleoside analogs. Evidence generated during the preceding period of support suggests that interruption of the PKC signal transduction pathway may promote cell death by three distinct mechanisms (a) dysregulation of cell cycle-related events, particularly induction of cyclin-dependent kinase inhibitors (CDKIs); (b) redirection of signals away from cytoprotective survival (e.g., MAPK/ERK) toward stress-related (e.g., SAPK/JNK) pathways; and (c) phosphorylation of the Bcl-2 protein, promoting mitochondrial permeability transition and circumvention of the block to capase activation. To examine these possibilities, (1) Human leukemic cells (HL-60, U937) stably overexpressing Bcl-2, Bcl-xL, and phosphorylation loop deletant mutants will be employed to determine whether phosphorylation of anti-apoptotic proteins is responsible for potentiation of ara-C mediated apoptosis by PKC inhibition/down-regulation, (2) Analogously, HL-60 and U937 cells expressing antisense p21WAF1+/-p27KIP1 will be used to characterize the effect of CDKI dysregulation on ara-C mediated apoptosis and its potentiation by PKC inhibitors/down-regulators, (3) The effects of enforced expression of p53 will be examined with respect to leukemic cell differentiation and modulation of ara-C induced cell death by bryostatin 1/PKC inhibitors, (4) Direct evidence for the participation of stress and survival pathways in cell death decisions will be obtained through the use of SEK1/JNK1 dominant-negative and ER-inducible Raf/MAPK mutants, (5) Findings will be extended to include another clinically important nucleoside analog, 2,2(1)-difluorodeoxcytidine (gemcitabine) and (6) Finally, the ability of pharmacologically relevant concentrations of bryostatin 1, UCN-01, and safingol to potentiate ara-C- (and gemcitabine)-mediated apoptosis in primary leukemic myeloblasts ex vivo will be explored. Information derived from these studies will lay the foundation for a novel approach to leukemia therapy aimed at enhancing the activity of effective antileukemic drugs via combination with agents that interrupt the PKC signal transduction pathway
Keywords: antileukemic agent, biological signal transduction, bryostatin, cytosine arabinoside, enzyme activity, myelogenous leukemia, programmed cell death, protein kinase C DNA damage, antisense nucleic acid, cell differentiation, cyclin dependent kinase, cytotoxicity, enzyme inhibitor, fludarabine, gene expression, mitogen activated protein kinase, mutant, myeloid stem cell, oncoprotein p21, pharmacokinetics, phosphorylation, staurosporine, tumor suppressor gene tissue /cell culture
Project start date: 1994-06-01
Project end date: 2003-04-30
3R01CA063753-07S1 (2001): $15835
3R01CA063753-05S1 (1999): $1
BRYOSTATIN 1 (NSC 339555) And FLUDARABINE IN LYMPHOCYTIC LEUKEMIA
Steven Grant, Professor
Virginia Commonwealth University Po Box 980568 Richmond, Va 232980568
Grant 5M01RR000065-380552 from National Center For Research Resources
Abstract: An investigation of the effects of using a new treatment of chronic lymphocytic leukemia and indolent Non-Hodgkin s lymphoma. A combination of Bryostatin 1, an experimental drug, and fludarabine, an established drug, will be given to 35 patients. The purpose of the study is to determine the safety, side effects, and correct doses of the combination in patients with symptomatic or advanced chronic lymphocytic leukemia (CLL) and relapsed, indolent (Non-Hodgkin s) lymphoma according to two sequences of administration. Also, to monitor apoptosis (fragmentation of a cell into membrane bound particles), differentiation, and a protein kinase C activity in leukemic lymphocytes exposed in vivo to bryostatin and fludarabine. And, to observe the anti-tumor activity of the combination of drugs in patients with CCL and relapse indolent lymphoma. Patients who have CCL or a previously treated lymphoma will receive fludarabine daily for five days and bryostatin for one day just before or after the fludarabine treatment. Order of the administration of the two drugs will be randomly determined and the doses will be measured by the experience of the patients previously treated. Dose increase will stop when serious side effects appear. Both drugs will be administered intravenously. Bryostatin will be continuously infused for 24 hours. Fludarabine will be infused for 30 minutes for five consecutive days. The treatment will be repeated every four weeks as long as the leukemia/lymphoma shrinks or remains stable and the side effects are acceptable. Progress will be checked during treatment by blood tests, physical exams, and other tests. Several extra blood samples will be used to determine how the drugs are acting in the body.
Keywords: acute lymphocytic leukemia, bryostatin, drug screening /evaluation, lymphoma, neoplasm /cancer chemotherapy, clinical trial phase I, dosage, drug adverse effect, human therapy evaluation, clinical research, human subject
Project start date: 1999-12-01
Project end date: 2000-11-30
Steven Grant
Virginia Commonwealth University
Project start date: 2003-05-01
Project end date: 2014-04-30
Sponsored Links Excellgen http://Excellgen.com