The present disclosure provides methods and compositions for microglia replacement therapy in a subject in need thereof. In some cases, the method involves administering myeloid cells to the central nervous system of a subject. In some cases, the myeloid cells are derived from embryonic or extraembryonic tissue. In some cases, the myeloid cells are genetically modified. The genetic modification may include a colony stimulating factor 1 receptor (CSF1R) variant that is resistant to a CSF1R inhibitor, yet retains sensitivity to its ligand (e.g., CSF1, IL34).

Microglia/monocytes exist within a ‘niche’ which limits the total number of microglia/monocytes/macrophages that reside within a mammalian central nervous system (CNS). Therefore, methods are needed that can help therapeutically modify microglia, monocytes, and macrophages or the cells that give rise to them to compete with endogenous microglia and partially or completely occupy the CNS niche. The present disclosure features therapeutic microglia, monocytes, or macrophages that have a selective advantage in comparison to endogenous brain resident microglia in their response to CSF1R inhibitors. Specifically, therapeutic cells developed in the present disclosure do not die at a given dose of CSF1R inhibitor that is sufficient to kill endogenous microglia. The therapeutic cells described herein can be used to treat neurological diseases.

Compositions and methods for the regulation of the cell cycle are encompassed, wherein the cell cycle is regulated via modulation of βcytokine activity.

A novel synthetic biology-based ADCC technology is provided that enhances or enables ADCC response. The novel ADCC technology can be used to prevent or treat cancers, infectious, inflammatory or autoimmune diseases, and other diseases where elimination of diseased cells is desirable.

A method of replacing endogenous microglia of a subject’s brain, e.g., an adult subject, with transplanted donor microglia includes depleting at least a portion of the endogenous microglia by administering to the subject a Colony Stimulating Factor 1 Receptor (CSFR1) inhibitor, wherein the CSFR1 inhibitor is blood-brain barrier permeable and pharmacologically ablates endogenous microglia; optionally stopping administration of the CSFR1 inhibitor for a time sufficient to prevent ablation of the transplanted donor microglia; and transplanting the donor microglia into the brain of the subject to provide the transplanted donor microglia.

Methods for Identifying protein modulators of eukaryotic cells by expressing a combinatorial library of potential agonists inside a eukaryotic cell and then directly selecting for an agonist of a target molecule. Some methods involve co-culturing a cell expressing a combinatorial library of potential agonists and a second cell, and then selecting agents that modulate a phenotype of or a desired cellular response in the second cell. Preferably, the agonists are antibodies introduced into and expressed in the starting cells, such as agonist anti-EpoR, anti-TpoR, or G-CSFR antibodies. Also disclosed are methods for selecting from combinatorial antibody libraries bispecific antibodies that can regulate cell phenotypes.

Disclosed is a group of chimeric antigen receptors (CARs) having two, three or four CAR molecules, wherein each member of the group of CARs is different in its amino acid sequence from one another, and wherein each of the CAR molecules of the group include at least a transmembrane domain and an ectodomain, wherein the ectodomain has one or two antigen binding moieties and/or one or two binding sites to which other polypeptides each including at least an antigen binding moiety are able to bind; wherein the ectodomain of each CAR molecule of the group in its prevalent conformation is free of cysteine amino acid moieties which are able to form intermolecular disulphide bonds with other CAR molecules of the group, respectively, and wherein each CAR molecule of the group includes at least one heterodimerization domain.

The present invention relates to a monoclonal antibody, or fragment thereof, which binds to CSF-1R (Colony stimulating factor 1 receptor), in particular to human CSF-1R. The present invention further relates to the in vitro use of the monoclonal antibody, or fragment thereof, of the present invention for the detection of CSF-1R in a sample. Further encompassed by the present invention is a complex comprising the monoclonal antibody, or fragment thereof, of the present invention and CSF-1R such as the human CSF-1R polypeptide.

Provided are a chimeric antigen receptor T lymphocyte for treating tumors, a preparation method therefor, and the use thereof. The chimeric antigen receptor successively comprises a single-chain antibody against a tumor cell surface antigen, a human hinge transmembrane region, a human intracellular signal domain, a self-cleaving peptide, and the full length of human CD27. The human intracellular signal domain comprises a human intracellular co-stimulatory signal domain and a human intracellular signaling domain. The method for preparing the chimeric antigen receptor T lymphocyte comprises the following steps: introducing the coding gene of the above-mentioned chimeric antigen receptor into a T cell and expressing the coding gene, thereby obtaining the CAR-T cell.

Methods and compositions are provided for reversibly localizing proteins to the exterior part of the cell surface. Compositions provided herein can include nucleic acids that encode polypeptides of interest and the ligand binding domain (LBD) of a nuclear hormone receptor. Medical applications are provided, including controlling the toxicity of CAR T cells.