Immunogenicity in the past year we have succeeded in identifying the major B cell epitopes recognized by the human immune system and have made a new immunotoxin, HA22-LR-010, that is highly active and whose reactivity with human anti-sera is greatly reduced. We are trying to further decrease reactivity by making further mutations in the new immunotoxin. We have also initiated a program to identify and remove human specific T cell epitopes. We have now identified all the major T cell epitopes in the toxin and using alanine scanning mutagenisis identified which amino acids within the epitope can be mutated to silence the epitope and maintain high cytotoxic activity. Using this information we have made 2 new types of recombinant immunotoxins. One of these has all the major T cell epitopes silenced. The other has a combination of mutations that remove B and T cell epitopes. These new RITs are now being tested in mice with human tumor xenographs. Based on our ability to remove B cell epitopes, Roche has produced a new immunotoxin targeting mesothelin which is now in preclinical development. In collaboration with David FitzGerald we have begun to study how protein synthesis arrest leads to apoptosis and have identified BAK, Bax, MCL-1 and BCL-XL as critical players in this process. We are also studying how protein phosphorylation regulates sensitivity of cells to immunotoxin mediated cell death by doing knock down studies of tyrosine and serine/threonine kinases and measuring the effect of knock down of these kinases on immunotoxin mediated cell death. We have identified the insulin receptor, HCK, BMX and the PDGF beta receptor as kinases that regulate immunotoxin killing. Lowing levels of these proteins enhances immunotoxin action. We have also identified several TK inhibitors that enhance immunotoxin action against cultured tumors cells and tumors growing in mice.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIABC008753-31
Application #
8763020
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
31
Fiscal Year
2013
Total Cost
$2,243,727
Indirect Cost
Name
National Cancer Institute Division of Basic Sciences
Department
Type
DUNS #
City
State
Country
Zip Code
Müller, Fabian; Cunningham, Tyler; Liu, Xiu Fen et al. (2016) Wide Variability in the Time Required for Immunotoxins to Kill B Lineage Acute Lymphoblastic Leukemia Cells: Implications for Trial Design. Clin Cancer Res 22:4913-4922
Mazor, Ronit; Onda, Masanori; Pastan, Ira (2016) Immunogenicity of therapeutic recombinant immunotoxins. Immunol Rev 270:152-64
Guha, Rajarshi; Mathews Griner, Lesley A; Keller, Jonathan M et al. (2016) Ranking Differential Drug Activities from Dose-Response Synthetic Lethality Screens. J Biomol Screen :
Liu, Xiufen; Müller, Fabian; Wayne, Alan S et al. (2016) Protein Kinase Inhibitor H89 Enhances the Activity of Pseudomonas Exotoxin A-Based Immunotoxins. Mol Cancer Ther 15:1053-62
Awuah, Prince; Bera, Tapan K; Folivi, Messan et al. (2016) Reduced Shedding of Surface Mesothelin Improves Efficacy of Mesothelin-Targeting Recombinant Immunotoxins. Mol Cancer Ther 15:1648-55
Illei, Peter B; Alewine, Christine; Zahurak, Marianna et al. (2016) Mesothelin Expression in Advanced Gastroesophageal Cancer Represents a Novel Target for Immunotherapy. Appl Immunohistochem Mol Morphol 24:246-52
Ali-Rahmani, Fatima; FitzGerald, David J; Martin, Scott et al. (2016) Anticancer Effects of Mesothelin-Targeted Immunotoxin Therapy Are Regulated by Tyrosine Kinase DDR1. Cancer Res 76:1560-8
Mazor, Ronit; Onda, Masanori; Park, Dong et al. (2016) Dual B- and T-cell de-immunization of recombinant immunotoxin targeting mesothelin with high cytotoxic activity. Oncotarget 7:29916-26
Bao, Xuhui; Chandramohan, Vidyalakshmi; Reynolds, Randall P et al. (2016) Preclinical toxicity evaluation of a novel immunotoxin, D2C7-(scdsFv)-PE38KDEL, administered via intracerebral convection-enhanced delivery in rats. Invest New Drugs 34:149-58
Mazor, Ronit; Tai, Chin-Hsien; Lee, Byungkook et al. (2015) Poor correlation between T-cell activation assays and HLA-DR binding prediction algorithms in an immunogenic fragment of Pseudomonas exotoxin A. J Immunol Methods 425:10-20

Showing the most recent 10 out of 102 publications