We propose the following paradigm for personalized anti-cancer therapy: a physician determines the unique surface-based characteristic (or """"""""fingerprint"""""""") of a cell type that he/she would like to target for elimination from a patient. Guided by the comparison of that fingerprint to the fingerprints of healthy cells, the physician combines off-the-shelf reagents and administers them to the patient, specifically killing that cell type (while protecting similar non-target cells and tissues). How realistic is this suggestion? Immunophenotype fingerprints for lymphomas and leukemias could be readily identified using monoclonal antibodies (MAb's). The topic of the current application is the second component of our paradigm: engineering modular off-the-shelf reagents that may be combined in order to trigger the death of target cells bearing these unique fingerprints, while protecting healthy cells at the same time. At the end of this funding period, we will provide proof-of-concept experiments for the complete paradigm, and either eliminate or protect any one or two out of three model cell lines (CCRF-CEM, Raji, HL-60), based on their immunophenotype fingerprints. It is our goal over the next three years to demonstrate that mixtures of molecules can perform AND and NOT logical operations on cell surfaces with therapeutically applicable outcomes (e.g., cell death or survival). We will first construct modular reagents that can be combined into oligonucleotide-Fab (antibody fragment) conjugates targeting cell- surface markers. In order to target selectively each of CCRF-CEM, Raji, and HL-60 cell lines, we will synthesize the reagents capable of targeting CD71, CD5, CD52, and CD33. The following specific aims will be then pursued:
Aim 1, AND Logic Demonstration: Selective oligonucleotide-based crosslinking of pairs of Fab-oligonucleotide conjugates on the surface of each one of the model cell lines. In this aim we will also study apoptotic/necrotic processes triggered by crosslinking with and without drug release.
Aim 2, NOT Logic Demonstration: Selective protection of each one of the model cell lines from the action of drug-oligonucleotide conjugate targeting markers found on all three cell lines. At the end of this funding period we will be ready to test this approach on actual samples from patients, and to initiate studies of self-assembly in murine models of leukemia and lymphoma. We will provide a proof-of-concept for the following paradigm for personalized anti- cancer therapy: a physician determines the unique surface-based characteristic (or """"""""fingerprint"""""""") of a cell type that he/she would like to target for elimination from a patient. Guided by the comparison of that fingerprint to the fingerprints of healthy cells, the physician combines off-the-shelf reagents and administers them to the patient, specifically killing that cell type (while protecting similar non-target cells and tissues). How realistic is this suggestion? Immunophenotype fingerprints for lymphomas and leukemias could be readily identified using monoclonal antibodies (MAb's). The topic of the current application is the second component of our paradigm: engineering modular off- the-shelf reagents that may be combined in order to trigger the death of target cells bearing these unique fingerprints, while protecting healthy cells at the same time. At the end of this funding period, we will provide proof-of-concept experiments for the complete paradigm, and either eliminate or protect any one or two out of three model cell lines, based on their immunophenotype fingerprints.
| Rudchenko, Maria; Taylor, Steven; Pallavi, Payal et al. (2013) Autonomous molecular cascades for evaluation of cell surfaces. Nat Nanotechnol 8:580-6 |
