The main reason why tumors are not controlled by the immune system of the cancer patient is that, unlike pathogens, tumors do not express potent tumor antigens that can be recognized by the immune system as "foreign". The current focus in developing immune-based modalities is to potentiate an immune response against the existing, albeit weak, antigens expressed in the tumor. An alternative approach - the focus of this proposal - is to express new, and hence potent, antigens in tumor cells in situ. Two main challenges that have precluded the development of such strategies, how to express new antigens in the disseminated tumor lesions of the cancer patients and how to restrict expression of such antigens to the tumor, precluding their expression in normal tissue, are addressed by the approach described in this proposal. Expression of novel antigens in tumor cells will be achieved using siRNA technology to inhibit nonsense mediated mRNA decay (NMD), a surveillance mechanism which prevents the expression of mRNAs containing a premature termination codon. Targeting siRNA inhibition to tumor cells - an essential requisite because of the constitutive nature and physiological roles of the NMD process - will be achieved using a novel targeting technology comprised of oligonucleotide aptamer ligands. Aptamers or aptamer targeted siRNA conjugates, unlike antibodies, can be synthesized in a chemical process, providing a more straightforward and cost effective manufacturing and regulatory approval process to generate clinical grade reagents. The hypotheses tested in this proposal - supported by extensive preliminary studies - are: (I). NMD inhibition in tumor cells will result in the expression of novel products which will function as tumor rejection antigens eliciting an immune response that will negatively impact on tumor growth, and (II). Aptamer-targeted siRNA inhibition of NMD in tumor-bearing mice is sufficiently robust to inhibit tumor growth. The main objectives of the proposed studies-carried out in preclinical murine models - are to provide proof-of-concept that tumor targeted inhibition of NMD using aptamer-siRNA conjugates will elicit tumor immunity, and to determine how effective is the this novel approach, compared to best-in-class "conventional" tumor vaccination protocols in preventing and reversing tumor progression in mice.
The specific aims of this proposal are: (1). To develop and characterize in vitro best-in-class PSMA aptamer targeted siRNAs corresponding to murine NMD-specific factors. (2). To evaluate the ability of PSMA aptamer-siRNA conjugates to engender antitumor immunity in tumor bearing mice. (3). To develop Her2 aptamer-siRNA conjugates and evaluate their ability to inhibit tumor growth in transplantable and spontaneous murine models for breast cancer. Successful completion of the studies proposed in this application will provide the rationale and guidelines for exploring aptamer targeted NMD inhibition to potentiate tumor immunogenicity in cancer patients.
Cancer vaccines aims at eliciting a potent antitumor immune responses capable of seeking out and destroying the disseminated tumor lesions in the cancer patient. The development of effective cancer vaccines is facing the challenges of identifying potent tumor antigens and effective "adjuvants" which are required to generate a robust immune response. Here we are proposing to develop an alternative, broadly applicable, and clinically feasible approach whereby new tumor antigens are expressed directly on the disseminated tumor cells, in effect "converting" tumor cells, immunologically, into pathogens.