The present disclosure relates to mRNA, self-replicating RNA, and temperature-sensitive, self-replicating RNA encoding a cancer antigen. The RNA constructs are suitable for cancer immunotherapy in a mammalian subject, such as a human subject.

The present invention provides improved and/or shortened processes and methods for preparing TILs in order to prepare therapeutic populations of TILs with increased therapeutic efficacy for the treatment of cancer with a KRAS mutation with TILs as described herein in combination with KRAS inhibitors.

The current disclosure provides for techniques and approaches for the generation of autologous mutant neoantigen-specific, TCR-engineered T cells used for adoptive transfer in treatment of cancer patients. Also provided are surrogate cancer cells, which is a personalized cell system that can be used for vaccination and TCR discovery in cancer patients.

Cancer stem cells (CSCs) are a subpopulation of tumor cells that can drive tumor initiation and can cause relapses. These cells are seen as drivers of tumor establishment and growth, often correlated to aggressive, heterogeneous and therapy-resistant tumors. Novel tumor-specific antigens (TSAs) and tumor-associated antigens (TAAs) specifically expressed by CSCs are described herein. Most of the TSAs described herein derives from aberrantly expressed unmutated genomic sequences, such as intronic and intergenic sequences, which are not expressed in normal tissues. Nucleic acids, compositions, cells, antibodies and vaccines derived from these TSAs are described. The use of the TSAs, nucleic acids, compositions, antibodies, cells and vaccines for the treatment of cancer, and more particularly cancers associated with the presence of CSCs such as poorly differentiated cancers, is also described.

An immunotherapeutic composition is contemplated that comprises subject-derived peripheral blood mononuclear cells (PBMC) and at least one recombinant adenovirus subtype 5 (Ad5) comprising a deletion in an E1 gene region, a deletion in an E2b gene region, and a nucleic acid sequence encoding a peptide antigen, wherein the PBMC are exposed ex-vivo to the at least one Ad5 vector. Advantageously, the same PBMC composition may also be used to prepare modified NK cells, and especially modified NK include CIML NK cells, CENK cells and mCENK cells.

The present invention relates to a method for identifying a truncal neo-antigen in a tumour from a subject which comprises the steps of: i) determining mutations present in a sample isolated from the tumour; and ii) identifying a truncal mutation which is a mutation present in essentially all tumour cells; and iii) identifying a truncal neo-antigen, which is an antigen encoded by a sequence which comprises the truncal mutation.

The invention relates to the field of cancer. In particular, it relates to the field of immune system directed approaches for tumor treatment, reduction and control. Some aspects of the invention relate to the identification of tumor specific neoantigens, such as those resulting from frameshift mutations, DNA rearrangements, or splicing mutations. Such neoantigens are useful for developing tumor treatments, such as vaccines or cellular immunotherapies and other means of stimulating a neoantigen specific immune response against a tumor in individuals.