Recombinant antibody T cell receptors (AbTCRs) and chimeric signaling receptors (CSRs) engineered to include antigen-binding domains specific for tumor antigen glypican-2 (GPC2) or glypican-3 (GPC3) are described. Immune cells expressing the AbTCRs and CSRs effectively treated GPC2-positive or GPC3-positive tumors in multiple different xenograft models, and were significantly more potent than similarly targeted chimeric antigen receptor (CAR) T cells.

Provided herein are antibodies and antigen-binding fragment thereof targeting GPC3, and chimeric antigen receptors having one or more anti-GPC3 antigen-binding fragments thereof. Further provided are engineered immune effector cells (e.g., T cells) expressing the chimeric antigen receptors and methods of use thereof.

Recombinant NK cells, and especially recombinant NK-92 cells express a chimeric antigen receptor (CAR) having an intracellular domain of FcRI. Notably, CAR constructs with an intracellular domain of FcRI had a substantially prolonged duration of expression and significantly extended cytotoxicity over time. The CAR may be expressed from RNA and DNA, preferably as a tricistronic construct that further encodes CD16 and a cytokine to confer autocrine growth support. Advantageously, such constructs also enable high levels of transfection and expression of the recombinant proteins and provide a convenient selection marker to facilitate rapid production of recombinant NK/NK-92 cells.

The present disclosure relates to the field of cell therapy, and more specifically, to improving CAR and/or TCR function through improvement of cytokine signaling. Disclosed are embodiments of a membrane bound interleukin 15 (IL-15)-IL-15Ralpha sushi domain chimeric receptor. The chimeric receptor may comprise a FAS transmembrane domain or an IL-7 transmembrane domain. Different embodiments of the chimeric receptor have different polypeptide lengths between the IL-15 region and the transmembrane domain.

The present invention provides a method for producing a natural killer cell from an iPS (Induced pluripotent stem) cell, comprising steps of: (i) contacting an iPS cell with a composition comprising a GSK-3 inhibitor and a ROCK inhibitor to obtain an embryoid body, (ii) contacting the embryoid body with a composition comprising a TGF receptor inhibitor to obtain a hematopoietic progenitor cell, (iii) culturing the hematopoietic progenitor cell to obtain a lymphocyte progenitor cell, and (iv) differentiating and expanding the lymphocyte progenitor cells to a natural killer cell.

The present application provides constructs comprising an antibody moiety specifically recognizing Glypican 3 (GPC3), such as a cell surface-bound GPC3. Also provided are methods of making and using these constructs.

Provided are a bispecific antibody or an antigen-binding fragment thereof, a nucleic acid encoding the same, a cell comprising the nucleic acid, a composition comprising the bispecific antibody or antigen-binding fragment thereof, the nucleic acid and/or the cell, and related applications of the bispecific antibody or antigen-binding fragment thereof in the treatment of cancer.

Monoclonal antibodies that bind glypican-2 (GPC2) with high affinity are described. Immunotoxins and chimeric antigen receptors (CARs) that include the disclosed antibodies or antigen-binding fragments thereof are further described. In some instances, the antibody or antigen-binding fragment is humanized. The disclosed GPC2-specific antibodies and conjugates can be used, for example, for the diagnosis or treatment of GPC2-positive cancers, including neuroblastoma, medulloblastoma and retinoblastoma.

Provided herein are novel Glypican 3 (GPC3) antibodies or antigen binding fragments and GPC3/CD3 bispecific antibodies. The present application also provides chimeric antigen receptors comprising the antibodies or antigen-binding fragments, related CAR-T cells, and preparation methods and uses of the same. The present application further provides pharmaceutical compositions comprising GPC3 antibodies or antigen binding fragments, related GPC3/CD3 bispecific antibodies, related GPC3 CAR or CAR-T cells, and methods of treating cancer in a subject in need thereof by administering the Glypican 3 (GPC3) antibodies or antigen binding fragments, the bispecific antibodies, the chimeric antigen receptors, the CAR-T cells, or the pharmaceutical compositions. The cancers treated in accordance with the application include Glypican-3-positive cancers.

Engineered T cells are provided that express (a) a chimeric antigen receptor wherein the antigen binding domain of the CAR binds to human GPC3 (a GPC-CAR); and (b) an orthogonal receptor. Also provided are methods of making and using such engineered T cells.