Mantle cell lymphoma (MCL) is a B-cell malignancy that is characterized by dysregulation of various oncogenes. Building on evidence that retinoic acid and its derivative, all trans retinoic acid (ATRA), are useful agents that potentiate apoptosis or anti-proliferative effects, it is proposed to pursue a strategy of targeted delivery of ATRA to MCL cells in culture. Using a novel delivery vehicle wherein ATRA is solubilized in nanoscale, protein stabilized lipid particles, termed nanodisks (ND), protein engineering methods will be employed to target ATRA-ND to the CD20 antigen present on the surface of B lymphocytes. This will be achieved by construction of a single chain variable antibody (scFv) apolipoprotein (apo) fusion protein. It is hypothesized that 1-CD20 scFv7apoA-I fusion protein will be capable of forming ND while retaining the antigen recognition properties of the parent scFv. Recombinant fusion protein will be expressed in E. coli, isolated and characterized. The ability of the chimera to associate with lipid and induce formation of ND will be determined while the antigen recognition properties of the 1-CD20 scFv portion of the fusion protein will be evaluated by Western blot and flow cytometry. ND prepared with wild type apoA-I will serve as control for these studies. In a second aim the effect of 1-CD20 scFv7apoA-I ATRA-ND on MCL cells in culture will be assessed. It is hypothesized that retinoid containing, targeted ND, will display enhanced induction of apoptosis in cell culture models of MCL. Different MCL cell lines will be employed in studies designed to evaluate targeting efficiency, concentration effectiveness and cell viability following exposure to CD20 targeted scFv7apoA-I ATRA-ND. Cultured cells will be exposed to control ATRA-ND and 1-CD20 scFv7apoA-I ATRA-ND followed by measurements of cell viability, apoptosis and autophagy. Dose- response and time course studies will be conducted to define optimal conditions. Studies will also be performed with ND harboring related synthetic and natural retinoids. The results of these studies will expand the potential of ND mediated drug delivery by demonstrating cell / tissue specific targeting, providing a framework for in vivo studies of targeted ND in animal models of MCL.
Public Health Relevance: Despite progress made in the broad category of lymphomas, mantle cell lymphoma (MCL) remains a poorly treated disease with median survival time of approximately 3 to 4 years. New therapy regimens have increased the complete remission rate but they have done little to change overall survival. Research proposed herein will evaluate the effectiveness of targeted drug payload delivery to cultured MCL cells. These studies will establish a novel approach with broad applicability, establishing targeted nanodisks as a platform that can be used with other forms of cancer, potentially decreasing the public burden associated with this disease.
Public Health Relevance Statement: Project narrative: Despite progress made in the broad category of lymphomas, mantle cell lymphoma (MCL) remains a poorly treated disease with median survival time of approximately 3 to 4 years. New therapy regimens have increased the complete remission rate but they have done little to change overall survival. Research proposed herein will evaluate the effectiveness of targeted drug payload delivery to cultured MCL cells. These studies will establish a novel approach with broad applicability, establishing targeted nanodisks as a platform that can be used with other forms of cancer, potentially decreasing the public burden associated with this disease.
NIH Spending Category: Bioengineering; Biotechnology; Cancer; Hematology; Lymphoma; Nanotechnology
Project Terms: (All-E)-3,7-Dimethyl-9-(2,6,6-trimethyl-1-cyclohexen-1-yl)-2,4,6,8-nonatetraenoic Acid; 9-cis-Retinoic Acid Receptor; ATGN; ATRA; Active Oxygen; Acute Promyelocytic Leukemia; Aggressive course; All-trans retinoic acid; Amino Acid Sequence; Animal Model; Animal Models and Related Studies; Antibodies; Antigens; Apo A-1; Apo A-I; Apo A1; Apo AI; ApoA-1; ApoA-I; Apolipoprotein A-1; Apolipoprotein A-I; Apolipoprotein A1; Apolipoprotein AI; Apolipoproteins; Apoptosis; Apoptosis Pathway; Autophagocytosis; B blood cells; B cell malignancy; B lymphoid malignancy; B-Cells; B-Lymphocyte Antigen CD20; B-Lymphocyte Surface Antigen B1; B-Lymphocytes; Binding; Binding (Molecular Function); Biological; Blotting, Western; Body Tissues; Bp35; Bursa-Dependent Lymphocytes; Bursa-Equivalent Lymphocyte; C-terminal; CD20; CD20 Antigens; CD20 Receptor; Cancer Genes; Cancer Treatment; Cancer-Promoting Gene; Cancers; Categories; Cell Culture Techniques; Cell Death, Programmed; Cell Line; Cell Lines, Strains; Cell Survival; Cell Viability; Cell-Death Protease; CellLine; Cells; Cellular Expansion; Cellular Growth; Chemosensitization; Chemosensitization/Potentiation; Chimera; Chimera Protein; Chimera organism; Chimeric Proteins; Code; Coding System; Controlled Study; Cultured Cells; Cytofluorometry, Flow; Disease; Disease remission; Disorder; Dose; Drug Delivery; Drug Delivery Systems; Drug Targeting; Drug Targetings; E coli; Effectiveness; Escherichia coli; Evaluation; Exhibits; Exposure to; Flow Cytofluorometries; Flow Cytometry; Flow Microfluorimetry; Forecast of outcome; Fusion Protein; Gene Targeting; Genetic Engineering of Proteins; Germinoblastoma; Human; Human, General; ICE-like protease; Induction of Apoptosis; Investigation; Investigators; Juvenile Chronic Myelogenous Leukemia; Juvenile Chronic Myeloid Leukemia; Juvenile Myelomonocytic Leukemia; Kaposi - Kaposi's Sarcoma; Kaposi Sarcoma; Kaposi?s Sarcoma; Leu-16; Lipids; Lymphoma; Lymphoma (Hodgkin's and Non-Hodgkin's); Lymphoma, Lymphocytic, Diffuse, Intermediate Differentiated; Lymphoma, Lymphocytic, Diffuse, Poorly-Differentiated; Lymphoma, Malignant; Lymphoma, Mantle-Cell; Lymphoma, Non-Hodgkin's; Lymphoma, Nonhodgkins; Lymphoma, Small-Cell, Centrocytic; MS4A1; MS4A1 gene; MS4A2; Malignant Neoplasm Therapy; Malignant Neoplasm Treatment; Malignant Neoplasms; Malignant Tumor; Malignant lymphoma, lymphocytic, intermediate differentiation, diffuse; Man (Taxonomy); Man, Modern; Mantle-Zone Lymphoma; Measurement; Mediating; Membrane-Spanning 4-Domains Subfamily A Member 1; Methods; Microfluorometry, Flow; Modeling; Molecular Interaction; Multiple Hemorrhagic Sarcoma; Myeloid Leukemia, Acute, M3; Neuroblastoma; Neuroblastoma (Schwannian Stroma-Poor); Non-Hodgkin's Lymphoma; Nuclear Hormone Receptor Superfamily; Nuclear Hormone Receptors; Oncogenes; Oxygen Radicals; Parents; Plasmid Cloning Vector; Plasmid Vector; Potentiation; Pro-Oxidants; Production; Prognosis; Progranulocytic Leukemia; Property; Property, LOINC Axis 2; Protein Engineering; Protein Structure, Primary; Proteins; Protocols, Treatment; RGM; RMSN; RXR; RXR Protein; Reaction Time; Reactive Oxygen Species; Recombinant Fusion Proteins; Regimen; Remission; Research; Research Design; Research Personnel; Researchers; Resistance; Response RT; Response Time; Reticulolymphosarcoma; Retinoic Acid; Retinoic Acid Agent; Retinoic Acid Receptor; Retinoic Acid Receptor RXR; Retinoic Acid and Derivatives; Retinoid X Receptors; Retinoids; Role; Sarcoma, Germinoblastic; Study Type; Surface; Targetings, Gene; Time; Tissues; Trans Vitamin A Acid; Transforming Genes; Treatment Protocols; Treatment Regimen; Treatment Schedule; Tretinoin; Tretinoinum; Vitamin A Acid; Western Blotting; Western Blottings; Western Immunoblotting; all-trans-Retinoic Acid; all-trans-Vitamin A acid; anticancer therapy; autophagy; base; bionano technology; bionanotechnology; cancer therapy; caspase; cell growth; cell type; cultured cell line; cystein protease; cystein proteinase; cysteine endopeptidase; disease/disorder; flow cytophotometry; gene product; high risk; immunogen; in vivo; juvenile chronic myelomonocytic leukemia; malignancy; member; model organism; nano biotechnology; nano meter scale; nano meter sized; nano scale; nanobiotechnology; nanometer scale; nanometer sized; nanoscale; neoplasm/cancer; new approaches; non-Hodgkin's lymphoma; non-Hodgkins disease; non-Hodgkins lymphoma; novel; novel approaches; novel strategies; novel strategy; outcome forecast; particle; promyelocytic leukemia; protein blotting; protein sequence; psychomotor reaction time; public health relevance; resistant; site targeted delivery; social role; study design; targeted delivery; trans-Retinoic Acid
