Cells and methods of preparation
AssigneeLIfT BioSciences Ltd
InventorsAlex Blyth, Oxana POLYAKOVA, Aoife MCGINLEY, Samuel FLORENCE, Mihil PATEL, Durva PATEL, Andrew WILLIS
Life of this patent
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Abstract
A method of preparing cells for therapeutic use is provided, in which a population of progenitor cells is cultured in cell culture conditions that promote differentiation of the progenitor cells comprising the presence of: G-CSF, GM-CSF, IL-3 and TNF; to produce a population of granulopoietic cells. The granulopoietic cells produced may be useful in methods of treatment, including the treatment of cancer and infections, and to modulate inflammatory cell responses.
Claims82
A method of preparing cells for therapeutic use, the method comprising:
- culturing a population of progenitor cells in cell culture conditions that promote differentiation of the progenitor cells comprising the presence of:
- G-CSF,
- GM-CSF,
- IL-3 and
- TNF;
- to produce a population of granulopoietic cells.
A method according to any preceding claim, wherein GM-CSF and IL-3 are provided to the cells for the final 48 hours of the period for which they are in culture.
A method according to any preceding claim, wherein TNF is provided to the cells for the final 24 hours of the period for which they are in culture.
A method according to any preceding claim, further comprising a step of culturing a population of stem cells in cell culture conditions to produce the population of progenitor cells:
- wherein the cell culture conditions for producing the progenitor cells comprise the presence of
- SCF,
- Flt-3 Ligand,
- IL-3,
- IL-6, and
- TPO.
A method according to any preceding claim, further comprising a step of purifying the population of granulopoietic cells produced.
A method according to any preceding claim, further comprising a step of formulating the population of granulopoietic cells produced for medical use.
A method according to any preceding claim, further comprising priming the granulopoietic cells for therapeutic use, by a method comprising culturing the population of granulopoietic cells in the presence of GM-CSF, and optionally one or more cytokines selected from the group consisting of:
- TNF, IFN-α, IFN-β, IL-15, and IL-18.
An isolated population of granulopoietic cells comprising:
- a first subpopulation of cells that are CD15+ CD64+ CD18+ CD49d+ CD71+;
- a second subpopulation of cells that are CD15− CD11b+/− CD18+ CD49d+ CD32+ HLA-DR−;
- and
- a third subpopulation of cells that are CD15− CD11b− HLA-DR+ CD18+ CD49d+ and CD71+.
A population of granulopoietic cells that are CD15+ CD64+ CD18+ CD49d+ CD71+.
A granulopoietic cell that is a CD64+ granulopoietic cell, or a population of such cells.
A granulopoietic cell that is a CD16− granulopoietic cell, or a population of such cells.
A CD62L− granulopoietic cell, or a population of such cells.
An isolated population of granulopoietic cells comprising:
- more than 90% Lin-cells (for example, approximately 97% Lin-cells);
- less than 30% CD34+ cells (for example, approximately 14% CD34+ cells);
- more than 30% CD38+ cells (for example, approximately 65% CD38+ cells);
- less than 1% cells with an HSC phenotype (for example approximately 0.04% cells with an HSC phenotype);
- less than 1% cells with an LT-HSC phenotype (for example approximately 0.02% cells with an LT-HSC phenotype;
- less than 20% cells with an LMPP phenotype (for example approximately 5% cells with an LMPP phenotype);
- and
- less than 10% cells with an MPP phenotype (for example approximately 2.5% cells with an MPP phenotype).
A method of treating a disease or disorder in a subject, the method comprising administering a pharmaceutical composition in accordance with the sixth aspect of the invention, or a population of granulopoietic cells in accordance with the second, or fourth to tenth, aspects of the invention, to the subject.
A method of treating cancer in a subject comprising administering a pharmaceutical composition of the invention to the subject.
Use of a pharmaceutical composition of the invention in the manufacture of a medicament for treating cancer in a subject.
A method of treating an infection in a subject comprising administering a pharmaceutical composition of the invention to the subject
Use of a pharmaceutical composition of the invention in the manufacture of a medicament for treating an infection in a subject.
A pharmaceutical composition of the invention, for use to amplify a non-granulocytic therapeutic immune response
A method of treatment comprising amplifying a non-granulocytic therapeutic immune response, the method comprising providing a pharmaceutical composition of the invention to a subject in need of such treatment
A pharmaceutical composition of the invention for use in the manufacture of a medicament for use in amplifying a non-granulocytic therapeutic immune response.
A composition comprising a population of granulopoietic cells as herein defined and a non-granulocytic immune cell.
A composition comprising a population of granulopoietic cells as herein defined and a non-granulocytic immune cell, wherein the composition does not comprise an αβ T cell
A composition comprising a granulopoietic cell as herein defined and a non-granulocytic immune cell, wherein the granulopoietic cell is capable of modulating (preferably modulates) the therapeutic immune response of the non-granulocytic immune cell.
A composition comprising a population of granulopoietic cells as herein defined and a non-granulocytic immune cell, wherein the population of granulopoietic cells is capable of amplifying (preferably amplifies) the therapeutic immune response of the non-granulocytic immune cell.
A kit comprising:
- (a) the composition according to the invention;
- or
- (b) a population of granulopoietic cells as herein defined and a non-granulocytic immune cell (e.g. a terminally differentiated non-granulocytic immune cell);
- and
- (c) optionally instructions for the use of the same (e.g. in treating cancer).
A method for manufacturing a composition (e.g. a composition of the invention), the method comprising:
- culturing PBMCs in the presence of granulopoietic cells as herein defined, thereby forming the composition;
- and optionally depleting αβ T cells before, during, or after the culturing.
A method for manufacturing a composition (e.g. a composition of the invention), the method comprising:
- culturing αβ T cell-depleted PBMCs under conditions that promote differentiation of progenitor cells present in the αβ T cell-depleted PBMCs into granulopoietic cells by a method as herein defined, thereby forming the composition.
A method of treating a disease or disorder in a subject comprising administering a composition of the invention to the subject.
A composition of the invention, for use to modulate a non-granulocytic therapeutic immune response.
A composition of the invention, for use to amplify a non-granulocytic therapeutic immune response.
A method of treatment comprising modulating a non-granulocytic therapeutic immune response, the method comprising providing a composition of the invention to a subject in need of such treatment.
A method of treatment comprising amplifying a non-granulocytic therapeutic immune response, the method comprising providing a composition of the invention to a subject in need of such treatment.
A composition of the invention for use in the manufacture of a medicament for use in modulating a non-granulocytic therapeutic immune response.
A composition of the invention for use in the manufacture of a medicament for use in amplifying a non-granulocytic therapeutic immune response.
A method of preparing a composition of the invention, the method comprising culturing a non-granulocytic immune cell in the presence of a population of granulopoietic cells of the invention.
A composition comprising a population of granulopoietic cells as herein defined and a non-granulocytic immune cell (e.g. a terminally differentiated non-granulocytic immune cell), wherein the population of granulopoietic cells is capable of amplifying (preferably amplifies) the therapeutic immune response of the non-granulocytic immune cell.
A composition comprising one or more granulocytes differentiated from a population of granulopoietic cells as herein defined capable of amplifying (preferably that amplifies) a therapeutic immune response of a non-granulocytic cell, and a non-granulocytic cell.
A method of treatment comprising modulating a non-granulocytic therapeutic immune response, the method comprising providing a composition of the invention to a subject in need of such treatment.
A method of treatment comprising amplifying a non-granulocytic therapeutic immune response, the method comprising providing a composition of the invention to a subject in need of such treatment.
A method of promoting therapeutic activity of non-granulocytic immune cells, the method comprising incubating a non-granulocytic immune cell with a population of granulopoietic cells as herein defined.
A method of increasing survival of immune cells in culture, the method comprising, culturing the immune cells in the presence of a feeder layer of granulopoietic cells as herein defined.
A method of increasing proliferation of immune cells in culture, the method comprising, culturing the immune cells in the presence of a feeder layer of granulopoietic cells as herein defined.
A method of selecting a suitable treatment regimen for a patient, the method comprising:
- identifying whether the patient has an impaired non-granulocytic immune response;
- and
- if the patient is identified as having an impaired non-granulocytic immune response, then treatment with a population of granulopoietic cells as herein defined is selected as an appropriate treatment;
- and
- if the patient is identified as lacking an impaired non-granulocytic immune response, then treatment with a therapy other than a population of granulopoietic cells as herein defined is selected.
A method of selecting a suitable treatment regimen for a patient, the method comprising:
- incubating a non-granulocytic immune cell from the patient with a population of granulopoietic cells as herein defined;
- wherein
- if the activation of the non-granulocytic immune cell from the patient is increased in response to the incubation, then treatment with a population of granulopoietic cells as herein defined is selected as an appropriate treatment;
- and
- if the activation of the non-granulocytic immune cell from the patient is increased in response to the incubation, then treatment with a therapy other than a population of granulopoietic cells as herein defined is selected.
A method of identifying whether or not a population of granulopoietic cells as herein defined is suitable for use in the treatment of cancer by beneficially modulating the tumour microenvironment, the method comprising:
- assessing whether the population of granulopoietic cells as herein defined, or a cell derived from the population of granulopoietic cells, is able to express proinflammatory cytokines;
- and/or
- assessing whether the population of granulopoietic cells as herein defined, or a cell derived from the population of granulopoietic cells, is able to stimulate expression of proinflammatory cytokines by non-granulocytic immune cells;
- and identifying whether or not a population of granulopoietic cells as herein defined is suitable for use in the treatment of cancer by beneficially modulating the tumour microenvironment on the basis of this assessment.
A method of identifying whether or not a population of granulopoietic cells as herein defined is suitable for use in the treatment of cancer by increasing recruitment of immune cells into a tumour and/or immune cell activation, the method comprising:
- assessing whether the population of granulopoietic cells as herein defined, or a cell derived from the population of granulopoietic cells, is able to express a chemokine associated with promoting cell trafficking;
- and/or
- assessing whether the population of granulopoietic cells as herein defined, or a cell derived from the population of granulopoietic cells, is able to stimulate expression of degranulation markers by non-granulocytic immune cells;
- and identifying whether or not a population of granulopoietic cells as herein defined is suitable for use in the treatment of cancer by increasing recruitment of immune cells into a tumour and/or immune cell activation on the basis of this assessment.
A method of identifying whether or not a population of granulopoietic cells as herein defined is suitable for use in the treatment of cancer by directly promoting killing of cancer cells, the method comprising:
- incubating the population of granulopoietic cells as herein defined, or a cell derived from the population of granulopoietic cells, with cells of a cancer cell line;
- and
- assessing whether the population of granulopoietic cells as herein defined, or a cell derived from the population of granulopoietic cells, is able to increase death of the cells of the cancer cell line to a greater extent than death of non-cancer cells;
- and identifying whether or not a population of granulopoietic cells as herein defined is suitable for use in the treatment of cancer by directly promoting killing of cancer cells on the basis of this assessment.
A method of identifying whether or not a population of granulopoietic cells as herein defined is suitable for use in the treatment of infection by directly promoting killing of cellular infectious agents or infected cells, the method comprising:
- incubating the population of granulopoietic cells as herein defined, or a cell derived from the population of granulopoietic cells, with a sample of a cellular infectious agent or of infected cells;
- and
- assessing whether the population of granulopoietic cells as herein defined, or a cell derived from the population of granulopoietic cells, is able to increase death of the cellular infectious agent or of infected cells;
- and identifying whether or not a population of granulopoietic cells as herein defined is suitable for use in the treatment of infection by directly promoting killing of cellular infectious agents or infected cells on the basis of this assessment.
A method of identifying whether or not a population of granulopoietic cells as herein defined is suitable for use in treatment by amplifying a therapeutic immune response, the method comprising:
- incubating the population of granulopoietic cells as herein defined, or a cell derived from the population of granulopoietic cells, with immune cells;
- and
- assessing whether the population of granulopoietic cells as herein defined, or the cell derived from the population of granulopoietic cells, is able to increase activation of the immune cells;
- and identifying whether or not a population of granulopoietic cells as herein defined is suitable for use in the treatment by amplifying a therapeutic immune response on the basis of this assessment.
Dependencies are inferred from the claim text.