NK cells with an increased antibody-dependent cellular toxicity (ADCC) against tumors
Inventors
Childs, Richard W. • Carlsten, Mattias C.V.
Assignees
US Department of Health and Human Services
Publication Number
US-10813952-B2
Publication Date
2020-10-27
Expiration Date
2035-11-13
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Abstract
Disclosed herein are modified NK cells, compositions comprising modified NK cells, and methods for treating a tumor or hyperproliferative disease in a subject. In some embodiments, the modified NK cells include NK cells including a heterologous nucleic acid molecule encoding a CD16 protein comprising a valine at amino acid position 158 (CD16-V158), a heterologous nucleic acid molecule encoding a CCR7 protein, or both. In some embodiments, methods include treating a subject with a tumor by administering a composition comprising an anti-cancer monoclonal antibody and administering a composition comprising the modified NK cells to the subject. Also disclosed are methods of making modified NK cells by obtaining a population of NK cells from a subject and transfecting the population of NK cells with a heterologous nucleic acid molecule encoding CD16-V158, a heterologous nucleic acid molecule encoding a CCR7 protein, or both.
Core Innovation
The invention discloses modified natural killer (NK) cells that include a heterologous nucleic acid molecule encoding a CD16 protein comprising a valine at amino acid position 158 (CD16-V158), a heterologous nucleic acid molecule encoding a CCR7 protein, or both. These modified NK cells, compositions comprising them, and methods for treating tumors or hyperproliferative diseases in subjects by administering such cells are provided. The methods further include treating a subject with a tumor by administering an anti-cancer monoclonal antibody and subsequently administering the modified NK cells to augment the anti-tumor immune response.
The background highlights the limitations of current NK cell therapies, including undesirable phenotypic changes following long-term culturing that can compromise homing capacity and cytotoxic function, as well as senescence compromising in vivo longevity. Moreover, genetic modification of NK cells via viral transduction is challenging due to substantial reductions in NK cell viability and low efficiency. Therefore, there is a need for methods to modify NK cells to enhance their in vivo viability, cytotoxicity, and homing ability to improve NK cell-based immunotherapy.
The invention addresses these challenges by utilizing mRNA electroporation to transiently transfect ex vivo expanded NK cells with nucleic acids encoding either the high-affinity CD16-V158 receptor, the CCR7 chemokine receptor to enhance homing to lymphoid tissues, or both. This provides modified NK cells with enhanced antibody-dependent cellular cytotoxicity (ADCC) against tumor cells and improved migration toward chemokine gradients. Transient expression through mRNA electroporation is efficient, maintains NK cell viability and cytotoxic function, and avoids the regulatory complexities and risks associated with viral transduction.
Claims Coverage
The independent claims focus on methods of treating multiple myeloma using modified NK cells.
Treatment method involving CD16-V158 modified NK cells with CD38 blockade
A method of treating a human subject with multiple myeloma comprising obtaining NK cells from the subject, transfecting or transducing the NK cells with a heterologous nucleic acid encoding a human CD16 protein with a valine at amino acid position 158 to create modified NK cells, blocking CD38 surface antigen on these modified NK cells using a Fab or F(ab)2 fragment of an anti-CD38 monoclonal antibody to produce CD38-blocked modified NK cells, administering an anti-CD38 antibody to the subject, and then administering the CD38-blocked modified NK cells to the subject.
Specific sequences encoding the CD16-V158 protein
The nucleic acid encoding the CD16 protein comprises the nucleic acid sequences of SEQ ID NO: 3 or SEQ ID NO: 5, and the encoded protein contains the amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 6, wherein valine at amino acid position 158 corresponds to amino acid 212 of SEQ ID NO: 4 or amino acid 176 of SEQ ID NO: 6.
Timing of administration for enhanced therapeutic effect
The composition comprising the CD38-blocked modified NK cells is administered to the subject approximately 1 to 8 hours after the administration of the anti-CD38 antibody composition.
Use of daratumumab as the anti-CD38 antibody
The anti-CD38 antibody administered to the subject in the method is daratumumab.
In vitro expansion prior to genetic modification
The population of NK cells is expanded in vitro prior to transfecting or transducing them with the heterologous nucleic acid encoding CD16-V158.
Administration sequence of antibody and modified NK cells
The composition comprising the modified NK cells is administered to the subject after administering the monoclonal antibody composition.
The independent claims cover methods of treating multiple myeloma by administering modified NK cells expressing CD16-V158 following in vitro expansion, where the modified cells have their CD38 blocked with antibody fragments, in combination with administration of an anti-CD38 antibody (such as daratumumab) to the subject. The claims emphasize nucleic acid and protein sequences encoding CD16-V158, timing of administration, and blocking of CD38 on NK cells to enhance therapy.
Stated Advantages
mRNA electroporation enables rapid, efficient, and non-toxic genetic modification of ex vivo expanded NK cells without using viral vectors.
Transient expression of CD16-V158 or CCR7 enhances NK cell antibody-dependent cellular cytotoxicity (ADCC) and homing ability without compromising proliferation, viability, or cytotoxic function.
Blocking CD38 on NK cells prior to administration prevents antibody-mediated NK cell depletion, enhancing therapeutic potential when combined with anti-CD38 antibodies like daratumumab.
The methods facilitate improved anti-tumor responses in subjects treated with monoclonal antibodies due to enhanced NK cell activity.
Documented Applications
Treatment of tumors and hyperproliferative diseases, including hematological malignancies such as multiple myeloma, chronic lymphocytic leukemia, acute lymphocytic leukemia, acute myeloid leukemia, Hodgkin's lymphoma, non-Hodgkin's lymphoma, mantle cell lymphoma, diffuse large B-cell lymphoma, and follicular lymphoma.
Treatment of solid tumors including lung carcinoma, prostate cancer, pancreatic cancer, breast cancer, colorectal adenocarcinoma, neuroblastoma, testicular cancer, and ovarian cancer.
Use of modified NK cells in combination with therapeutic monoclonal antibodies such as daratumumab for multiple myeloma and rituximab for B cell lymphomas.
Enhancing NK cell homing to lymphoid tissues to improve treatment efficacy against lymphomas or metastases in lymphoid tissues by modifying NK cells to express CCR7.
Interested in licensing this patent?