Genetically engineered cells having multi-transmembrane domain chimeric antigen receptors utilizing g protein-coupled receptor scaffolds, and uses thereof

Abstract

Provided are iPSCs and derivative cells thereof expressing multi-transmembrane domain chimeric antigen receptors utilizing G protein-coupled receptor (GPCR) scaffolds, and uses of the iPSCs or the derivative cells to express a GPCR chimeric antigen receptor (CAR) for allogenic cell therapy.

Description

[0001] Docket No.: 066461.11126 / 25WO1 GENETICALLY ENGINEERED CELLS HAVING MULTI-TRANSMEMBRANE DOMAIN CHIMERIC ANTIGEN RECEPTORS UTILIZING G PROTEIN-COUPLED RECEPTOR SCAFFOLDS, AND USES THEREOF REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Patent Application No.63 / 597,762 filed November 10, 2023, the disclosure of which is incorporated herein by reference in its entireties. TECHNICAL FIELD This application provides genetically engineered induced pluripotent stem cells (iPSCs) and derivative cells having multi-transmembrane domain chimeric antigen receptors utilizing G protein-coupled receptor (GPCR) scaffolds. Also provided are uses of the iPSCs or derivative cells thereof to express a GPCR chimeric antigen receptor (CAR) for allogenic cell therapy. Also provided are related vectors, polynucleotides, and pharmaceutical compositions. REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY This application contains a sequence listing, which is submitted electronically as an XML formatted sequence listing with a file name “066461-25WO1 Sequence Listing.xml” having a file size of 389 kilobytes, and a creation date of October 28, 2024. The sequence listing submitted via Patent Center is part of the specification and is herein incorporated by reference in its entirety. INCORPORATION BY REFERENCE All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. Docket No.: 066461.11126 / 25WO1 BACKGROUND Conventional CARs consist of an antigen-binding domain (e.g., derived from an antibody or antibody-like molecule), a flexible hinge / spacer, a single transmembrane domain (e.g., derived from CD28 or CD8), and intracellular domains that incorporate the intracellular domain of CD3z and one or more costimulatory domains (e.g., of CD28 or 4-1BB / CD137). While the extracellular domain of the CAR engages the target antigen and allows for the formation of an immune synapse, the intracellular domains of the CAR transduce immune effector signals from the intracellular domains and lead to activation, proliferation, cytokine secretion, and cytotoxicity against the target antigen-bearing cell. Research on CARs in recent years has focused on engineering, optimizing, and adding functionality into the receptors. Multiple antigen specificities can be engineered into the CAR extracellular domain by incorporating additional binders, or additional or better effector responses can be conferred to CAR T or NK cells by adding intracellular domains from other immune receptors (e.g., DAP10, ICOS, DNAM1, and LAT). However, since traditional CARs have a single transmembrane domain, any additional intra- or extracellular domains must be built onto the receptor in a linear fashion. For extracellular domains, this increases the distance of the binding moieties from the effector cell membrane, which can reduce CAR functionality. Similarly, for intracellular domains, increased distance from the cell membrane may reduce accessibility of those domains to membrane-bound kinases, such as lymphocyte-specific protein tyrosine kinase (Lck), which are partly involved in transducing the effector response. Current alternatives to these approaches include the engineering of additional receptors beyond the primary CAR that incorporate alternative intracellular domains, or engineering an additional CAR in tandem with the primary CAR. While expressing multiple receptors can be a route to greater efficacy, it also presents challenges. For iPSC-derived allogeneic cell therapies in which transgenes are introduced via CRISPR gene editing, engineering multiple receptors involves additional gene editing loci that must maintain robust expression throughout the differentiation process. Alternatively, one could use a single locus, multi-cistronic system (e.g., 2A), however, expression of each coding sequence can be inconsistent. In the context of autologous cell therapies, in which the CAR is transduced into effector cells by a viral Docket No.: 066461.11126 / 25WO1 vector, the packaging size of the transgene is limiting, with too large DNA cassettes inhibiting viral production and transduction. These challenges with multiple receptor engineering also preclude the exploration of further receptors beyond two, due to the same challenges. To address these challenges, the present disclosure provides iPSCs and derivative cells that utilize G Protein-Coupled Receptors (GPCRs) as a protein scaffold for the incorporation of additional intra- and extracellular domains by engineering the intracellular and extracellular loops of GPCRs to carry the costimulatory or activating domains of immune effector receptors (as intracellular domains), or antigen binding moieties such as VHH and scFv (as extracellular domains). BRIEF DESCRIPTION OF THE DRAWINGS The foregoing summary, as well as the following detailed description of preferred embodiments of the present application, will be better understood when read in conjunction with the appended drawings. It should be understood, however, that the application is not limited to the precise embodiments shown in the drawings. FIG.1 shows various CAR constructs (utilizing GPCR scaffolds and targeting an EGFR antigen) which are used in certain embodiments of the present disclosure. CAR extracellular domains were engineered with anti-EGFR VHH binders. Selected CARs were expressed and tested for cytotoxicity activity in T-cells against EGFR-expressing Nalm6 tumor cells. FIG.2A-B shows (A) the expression of various GPCR CARs of the present disclosure in lentivirus-transduced Nurkat cells; and (B) AF647 positive Nurkat cells transduced with various GPCR CARs of the present disclosure. FIG.3 shows the expression of various GPCR CARs of the present disclosure in lentivirus-transduced primary T cells. FIG.4 shows the expression of various GPCR CARs of the present disclosure in lentivirus-transduced primary T cells. FIG.5 shows EGFR-dependent killing of Nalm6 cells using therapeutic cells expressing monospecific anti-EGFR GPCR CAR constructs of the present disclosure at a 1:5 E:T ratio. Docket No.: 066461.11126 / 25WO1 FIG.6A-B shows (A) a schematic of downstream signaling from conventional CARs (left) and (B) CARs utilizing a chemokine GPCR scaffold. DETAILED DESCRIPTION Various publications, articles and patents are cited or described in the background and throughout the specification; each of these references is herein incorporated by reference in its entirety. Discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is for the purpose of providing context for the invention. Such discussion is not an admission that any or all of these matters form part of the prior art with respect to any inventions disclosed or claimed. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this application pertains. Otherwise, certain terms used herein have the meanings as set forth in the specification. It must be noted that as used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural reference unless the context clearly dictates otherwise. Unless otherwise stated, any numerical values, such as a concentration or a concentration range described herein, are to be understood as being modified in all instances by the term “about.” Thus, a numerical value typically includes ± 10% of the recited value. For example, a concentration of 1 mg / mL includes 0.9 mg / mL to 1.1 mg / mL. Likewise, a concentration range of 1% to 10% (w / v) includes 0.9% (w / v) to 11% (w / v). As used herein, the use of a numerical range expressly includes all possible subranges, all individual numerical values within that range, including integers within such ranges and fractions of the values unless the context clearly indicates otherwise. Unless otherwise indicated, the term “at least” preceding a series of elements is to be understood to refer to every element in the series. Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the application described herein. Such equivalents are intended to be encompassed by the application. Docket No.: 066461.11126 / 25WO1 As used herein, the terms “comprises,” “comprising,” “includes,” “including,” “has,” “having,” “contains” or “containing,” or any other variation thereof, will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers and are intended to be non-exclusive or open-ended. For example, a composition, a mixture, a process, a method, an article, or an apparatus that comprises a list of elements is not necessarily limited to only those elements but can include other elements not expressly listed or inherent to such composition, mixture, process, method, article, or apparatus. Further, unless expressly stated to the contrary, “or” refers to an inclusive or and not to an exclusive or. For example, a condition A or B is satisfied by any one of the following: A is true (or present), and B is false (or not present), A is false (or not present) and B is true (or present), and both A and B are true (or present). As used herein, the conjunctive term “and / or” between multiple recited elements is understood as encompassing both individual and combined options. For instance, where two elements are conjoined by “and / or,” a first option refers to the applicability of the first element without the second. A second option refers to the applicability of the second element without the first. A third option refers to the applicability of the first and second elements together. Any one of these options is understood to fall within the meaning, and therefore satisfy the requirement of the term “and / or” as used herein. Concurrent applicability of more than one of the options is also understood to fall within the meaning, and therefore satisfy the requirement of the term “and / or.” As used herein, the term “consists of,” or variations such as “consist of” or “consisting of,” as used throughout the specification and claims, indicate the inclusion of any recited integer or group of integers, but that no additional integer or group of integers can be added to the specified method, structure, or composition. As used herein, the term “consists essentially of,” or variations such as “consist essentially of” or “consisting essentially of,” as used throughout the specification and claims, indicate the inclusion of any recited integer or group of integers, and the optional inclusion of any recited integer or group of integers that do not materially change the basic or novel properties of the specified method, structure or composition. See M.P.E.P. § 2111.03. Docket No.: 066461.11126 / 25WO1 As used herein, “subject” means any animal, preferably a mammal, most preferably a human. The term “mammal” as used herein, encompasses any mammal. Examples of mammals include, but are not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc., more preferably a human. It should also be understood that the terms “about,” “approximately,” “generally,” “substantially,” and like terms, used herein when referring to a dimension or characteristic of a component of the preferred invention, indicate that the described dimension / characteristic is not a strict boundary or parameter and does not exclude minor variations therefrom that are functionally the same or similar, as would be understood by one having ordinary skill in the art. At a minimum, such references that include a numerical parameter would include variations that, using mathematical and industrial principles accepted in the art (e.g., rounding, measurement or other systematic errors, manufacturing tolerances, etc.), would not vary the least significant digit. The term "chimeric antigen receptor” or “CAR” refers to engineered receptors, which are grafted onto cells. In general, a CAR of the present disclosure comprises one or more extracellular domains comprising the antigen binding domain(s), one or more intracellular domains comprising one or more costimulatory and / or signaling domains, and a scaffold comprising multiple transmembrane domains and intracellular or extracellular loops, at which the one or more extracellular or intracellular domains are disposed. The antigen binding domain of the CAR targets specific antigens. The targeting regions may comprise full length heavy chain, Fab fragments, scFvs, divalent single chain antibodies or diabodies, each of which are specific to the target antigen (e.g., a tumor antigen of Table 1). The antigen binding domain can be derived from the same species or a different species for or in which the CAR will be used in. The term "transmembrane domain" refers to the region of the CAR, which crosses or bridges the plasma membrane. In certain embodiments of the present disclosure, the CAR scaffold comprises a GPCR, which can multiple (e.g., two, three, four, five, or more) transmembrane domains. As used herein, the term "G protein-coupled receptor" or "GPCR," or "GPR" refers to a transmembrane receptor that is capable of transmitting a signal from the outside of a cell to the inside of a cell through a G protein pathway and / or an arrestin Docket No.: 066461.11126 / 25WO1 pathway. Hundreds of such receptors are known in the art; see, e.g., Fredriksson et al., Mol. Pharmacol.63: 1256-1272, 2003, and Vassilatis, D.K., Proc Natl Acad Sci USA 100: 4903-4908 (2003), each of which are hereby incorporated by reference. These references have characterized the human and mouse GPCRs based on sequence homology and function. Human GPCRs can be broken down into five classes: secretin, rhodopsin, glutamate, frizzled / Tas2, and adhesion. Alternatively, receptors may be classified by their ligands, e.g., peptide hormones or small molecules (e.g., biogenic amines). Other classification schemes include the A-F classification, where class A represents receptors related to rhodopsin and the adrenergic receptors, class B, receptors related to the calcitonin and parathyroid hormone receptors, class C, receptors related to the metabotropic receptors, and classes D-F represent receptors found in fungi and archaebacteria. In certain embodiments of the present disclosure, GPCRs can be used as a scaffold for a CAR. The term "G protein" refers to a heterotrimeric protein complex that transmits a signal from an activated GPCR to effector molecule(s) inside the cell such as enzymes and ion channels. G proteins are made up of Got, Gp, and Gy subunits. Families of Got subunits include Gq, Gi, Gs, and Gal2 / 13. G protein signaling pathways are named for the activated Got subunit, / .< ., Gas, Gai, Gaq, and Gal 2 / 13. A heterotrimeric G protein binds to an activated GPCR protein, that is, a GPCR protein that is bound to a ligand or surrogate ligand. When bound to a GPCR protein, the Ga subunit exchanges bound guanosine diphosphate (GDP) for guanosine-5'-triphosphate (GTP) and dissociates from the GP and Gy subunits, which are typically associated in a heterodimeric complex. Once dissociated, both the Ga-GTP -bound protein and the GPy complex can activate signaling pathways. The Gq family includes Gaq, Gal 1, Gal4, and Gal5 / 16. The Gi family includes Gail-3, Gao, Gat, Gagust, and Gaz. The Gs family includes Gas and Gaolf. The G12 / 13 includes Gal2 and Gal3. The terms “binder” or “specifically binds” or “specific for” with respect to an antigen-binding domain of a ligand like an antibody, of a fragment thereof or of a CAR refer to an antigen-binding domain which recognizes and binds to a specific antigen, but does not substantially recognize or bind other molecules in a sample. An antigen-binding domain that binds specifically to an antigen from one species may bind also to that Docket No.: 066461.11126 / 25WO1 antigen from another species. This cross-species reactivity is not contrary to the definition of that antigen-binding domain as specific. An antigen-binding domain that specifically binds to an antigen may bind also to different allelic forms of the antigen (allelic variants, splice variants, isoforms etc.). This cross reactivity is not contrary to the definition of that antigen-binding domain as specific. The terms “engineered cell” and “genetically modified cell” as used herein can be used interchangeably. The terms mean containing and / or expressing a foreign gene or nucleic acid sequence which in turn modifies the genotype or phenotype of the cell or its progeny. Especially, the terms refers to cells, preferentially T cells which are manipulated by recombinant methods well known in the art to express stably or transiently peptides or proteins which are not expressed in these cells in the natural state. For example, T cells are engineered to express an artificial construct such as a chimeric antigen receptor on their cell surface. For example, the sequences encoding the CAR may be delivered into cells using a retroviral or lentiviral vector. The term “primary cell” refers to a cell isolated directly from a multicellular organism. Primary cells typically have undergone very few population doublings and are therefore more representative of the main functional component of the tissue from which they are derived in comparison to continuous (tumor or artificially immortalized) cell lines. In some cases, primary cells are cells that have been isolated and then used immediately. In other cases, primary cells cannot divide indefinitely and thus cannot be cultured for long periods of time in vitro. In certain embodiments of the present disclosure, a primary cell can comprise a primary immune cell (e.g., a primary T-cell) The term “target” as used herein refers to an antigen or epitope associated with a cell that should be recognized specifically by an antigen binding domain, e.g. an antigen binding domain of an antibody or of a CAR. The antigen or epitope for antibody recognition can be bound to the cell surface but also be secreted, part of the extracellular membrane, or shed from the cell. As used herein, the term “dual-targeting” refers to a protein (e.g., a chimeric protein) capable of binding to two different antigens. Specifically, a dual-targeting protein of the present disclosure (e.g., a CAR having two or more tumor or cancer antigen binding domains) does not naturally occur and is produced by a genetic engineering Docket No.: 066461.11126 / 25WO1 method or other method. In one embodiment, a primary cell, an engineered iPSC or derivative cell of the present disclosure can comprise one or more exogenous polynucleotides encoding a CAR having a first antigen binding domain that specifically binds CD19 and a second antigen binding domain that specifically binds CD22. This is in contrast with other examples of the present disclosure wherein a primary cell, an engineered iPSC or derivative cell comprises one or more polynucleotides encoding a first CAR having a first antigen binding domain that specifically binds CD33 and a second CAR having a second antigen binding domain that specifically binds CD123. The terms “identical” or percent “identity,” in the context of two or more nucleic acids or polypeptide sequences (e.g., CAR polypeptides and the CAR polynucleotides that encode them), refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same, when compared and aligned for maximum correspondence, as measured using one of the following sequence comparison algorithms or by visual inspection. For sequence comparison, typically one sequence acts as a reference sequence, to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are input into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. The sequence comparison algorithm then calculates the percent sequence identity for the test sequence(s) relative to the reference sequence, based on the designated program parameters. Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith & Waterman, Adv. Appl. Math.2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol.48:443 (1970), by the search for similarity method of Pearson & Lipman, Proc. Nat’l. Acad. Sci. USA 85:2444 (1988), by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, WI), or by visual inspection (see generally, Current Protocols in Molecular Biology, F.M. Ausubel et al., eds., Current Protocols, a joint venture between Greene Publishing Associates, Inc. and John Wiley & Sons, Inc., (1995 Supplement) (Ausubel)). Docket No.: 066461.11126 / 25WO1 Examples of algorithms that are suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al. (1990) J. Mol. Biol.215: 403-410 and Altschul et al. (1997) Nucleic Acids Res.25: 3389-3402, respectively. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information. This algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence, which either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul et al., supra). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are then extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always > 0) and N (penalty score for mismatching residues; always < 0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a wordlength (W) of 11, an expectation (E) of 10, M=5, N= -4, and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a wordlength (W) of 3, an expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff & Henikoff, Proc. Natl. Acad. Sci. USA 89:10915 (1989)). In addition to calculating percent sequence identity, the BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, e.g., Karlin & Altschul, Proc. Nat’l. Acad. Sci. USA 90:5873-5787 (1993)). One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability by which a match between two Docket No.: 066461.11126 / 25WO1 nucleotide or amino acid sequences would occur by chance. For example, a nucleic acid is considered similar to a reference sequence if the smallest sum probability in a comparison of the test nucleic acid to the reference nucleic acid is less than about 0.1, more preferably less than about 0.01, and most preferably less than about 0.001. A further indication that two nucleic acid sequences or polypeptides are substantially identical is that the polypeptide encoded by the first nucleic acid is immunologically cross reactive with the polypeptide encoded by the second nucleic acid, as described below. Thus, a polypeptide is typically substantially identical to a second polypeptide, for example, where the two peptides differ only by conservative substitutions. Another indication that two nucleic acid sequences are substantially identical is that the two molecules hybridize to each other under stringent conditions. As used herein, the term “isolated” means a biological component (such as a nucleic acid, peptide, protein, or cell) has been substantially separated, produced apart from, or purified away from other biological components of the organism in which the component naturally occurs, i.e., other chromosomal and extrachromosomal DNA and RNA, proteins, cells, and tissues. Nucleic acids, peptides, proteins, and cells that have been “isolated” thus include nucleic acids, peptides, proteins, and cells purified by standard purification methods and purification methods described herein. “Isolated” nucleic acids, peptides, proteins, and cells can be part of a composition and still be isolated if the composition is not part of the native environment of the nucleic acid, peptide, protein, or cell. The term also embraces nucleic acids, peptides and proteins prepared by recombinant expression in a host cell as well as chemically synthesized nucleic acids. As used herein, the term “polynucleotide,” synonymously referred to as “nucleic acid molecule,” “nucleotides” or “nucleic acids,” refers to any polyribonucleotide or polydeoxyribonucleotide, which can be unmodified RNA or DNA or modified RNA or DNA. “Polynucleotides” include, without limitation single- and double-stranded DNA, DNA that is a mixture of single- and double-stranded regions, single- and double- stranded RNA, and RNA that is mixture of single- and double-stranded regions, hybrid molecules comprising DNA and RNA that can be single-stranded or, more typically, double-stranded or a mixture of single- and double-stranded regions. In addition, Docket No.: 066461.11126 / 25WO1 “polynucleotide” refers to triple-stranded regions comprising RNA or DNA or both RNA and DNA. The term polynucleotide also includes DNAs or RNAs containing one or more modified bases and DNAs or RNAs with backbones modified for stability or for other reasons. “Modified” bases include, for example, tritylated bases and unusual bases such as inosine. A variety of modifications can be made to DNA and RNA; thus, “polynucleotide” embraces chemically, enzymatically or metabolically modified forms of polynucleotides as typically found in nature, as well as the chemical forms of DNA and RNA characteristic of viruses and cells. “Polynucleotide” also embraces relatively short nucleic acid chains, often referred to as oligonucleotides. A “construct” refers to a macromolecule or complex of molecules comprising a polynucleotide to be delivered to a host cell, either in vitro or in vivo. A “vector,” as used herein refers to any nucleic acid construct capable of directing the delivery or transfer of a foreign genetic material to target cells, where it can be replicated and / or expressed. The term “vector” as used herein comprises the construct to be delivered. A vector can be a linear or a circular molecule. A vector can be integrating or non-integrating. The major types of vectors include, but are not limited to, plasmids, episomal vector, viral vectors, cosmids, and artificial chromosomes. Viral vectors include, but are not limited to, adenovirus vector, adeno-associated virus vector, retrovirus vector, lentivirus vector, Sendai virus vector, and the like. By “integration” it is meant that one or more nucleotides of a construct is stably inserted into the cellular genome, i.e., covalently linked to the nucleic acid sequence within the cell's chromosomal DNA. By “targeted integration” it is meant that the nucleotide(s) of a construct is inserted into the cell's chromosomal or mitochondrial DNA at a pre-selected site or “integration site”. The term “integration” as used herein further refers to a process involving insertion of one or more exogenous sequences or nucleotides of the construct, with or without deletion of an endogenous sequence or nucleotide at the integration site. In the case, where there is a deletion at the insertion site, “integration” can further comprise replacement of the endogenous sequence or a nucleotide that is deleted with the one or more inserted nucleotides. As used herein, the term “exogenous” is intended to mean that the referenced molecule or the referenced activity is introduced into, or non-native to, the host cell. The Docket No.: 066461.11126 / 25WO1 molecule can be introduced, for example, by introduction of an encoding nucleic acid into the host genetic material such as by integration into a host chromosome or as non- chromosomal genetic material such as a plasmid. Therefore, the term as it is used in reference to expression of an encoding nucleic acid refers to introduction of the encoding nucleic acid in an expressible form into the cell. The term “endogenous” refers to a referenced molecule or activity that is present in the host cell in its native form. Similarly, the term when used in reference to expression of an encoding nucleic acid refers to expression of an encoding nucleic acid natively contained within the cell and not exogenously introduced. As used herein, a “gene of interest” or “a polynucleotide sequence of interest” is a DNA sequence that is transcribed into RNA and in some instances translated into a polypeptide in vivo when placed under the control of appropriate regulatory sequences. A gene or polynucleotide of interest can include, but is not limited to, prokaryotic sequences, cDNA from eukaryotic mRNA, genomic DNA sequences from eukaryotic (e.g., mammalian) DNA, and synthetic DNA sequences. For example, a gene of interest may encode an miRNA, an shRNA, a native polypeptide (i.e. a polypeptide found in nature) or fragment thereof; a variant polypeptide (i.e. a mutant of the native polypeptide having less than 100% sequence identity with the native polypeptide) or fragment thereof; an engineered polypeptide or peptide fragment, a therapeutic peptide or polypeptide, an imaging marker, a selectable marker, and the like. “Operably-linked” refers to the association of nucleic acid sequences on a single nucleic acid fragment so that the function of one is affected by the other. For example, a promoter is operably-linked with a coding sequence or functional RNA when it is capable of affecting the expression of that coding sequence or functional RNA (i.e., the coding sequence or functional RNA is under the transcriptional control of the promoter). Coding sequences can be operably-linked to regulatory sequences in sense or antisense orientation. The term “expression” as used herein, refers to the biosynthesis of a gene product. The term encompasses the transcription of a gene into RNA. The term also encompasses translation of RNA into one or more polypeptides, and further encompasses all naturally occurring post-transcriptional and post-translational modifications. The expressed CAR Docket No.: 066461.11126 / 25WO1 can be within the cytoplasm of a host cell, into the extracellular milieu such as the growth medium of a cell culture or anchored to the cell membrane. As used herein, the terms “peptide,” “polypeptide,” or “protein” can refer to a molecule comprised of amino acids and can be recognized as a protein by those of skill in the art. The conventional one-letter or three-letter code for amino acid residues is used herein. The terms “peptide,” “polypeptide,” and “protein” can be used interchangeably herein to refer to polymers of amino acids of any length. The polymer can be linear or branched, it can comprise modified amino acids, and it can be interrupted by non-amino acids. The terms also encompass an amino acid polymer that has been modified naturally or by intervention; for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification, such as conjugation with a labeling component. Also included within the definition are, for example, polypeptides containing one or more analogs of an amino acid (including, for example, unnatural amino acids, etc.), as well as other modifications known in the art. The peptide sequences described herein are written according to the usual convention whereby the N-terminal region of the peptide is on the left and the C-terminal region is on the right. Although isomeric forms of the amino acids are known, it is the L- form of the amino acid that is represented unless otherwise expressly indicated. As used herein, the term “engineered immune cell” refers to an immune cell, also referred to as an immune effector cell, that has been genetically modified by the addition of exogenous genetic material in the form of DNA or RNA to the total genetic material of the cell. Overview The present disclosure provides engineered cells (and compositions comprising such cells) having CARs that utilize a GPCR scaffold to enable additional functional extracellular and intracellular domains to be engineered into a single receptor. Generally, GPCRs contain 7 transmembrane domains, 3 intracellular and extracellular loops, an extracellular N-terminal region, and intracellular C-terminal tail. By engineering (i) intracellular domains on different intracellular loops (or the C terminus), and / or (ii) antigen binders on the N terminus or extracellular loops, multiple functional extracellular Docket No.: 066461.11126 / 25WO1 and intracellular domains can be engineered into a single CAR. Additionally, the binders and endodomains incorporated into intracellular or extracellular loops are maintained proximal to the cell membrane, which may help synapse formation and receptor functionality for each domain. GPCRs are amenable to engineering, as the loops and termini of GPCRs are flexible and do not contribute to the overall structural framework of the GPCR. Finally, chemokine receptors such as CXCR3 and CXCR4 are expressed on T and NK cells and natively contribute to the effector response. CARs that utilize these receptors as a scaffold may signal through endogenous pathways and further increase functionality. A single receptor with one or more tumor antigen specificities can be designed that also recapitulate multiple modes of immune effector signaling, providing more robust T or NK responses, persistence, and cytokine release. In certain aspects, the present disclosure provides an induced pluripotent stem cell (iPSC) or a derivative cell thereof comprising at least one exogenous polynucleotide encoding one or more chimeric antigen receptors (CARs), comprising a G protein- coupled receptor (GPCR) scaffold or fragment thereof and at least one antigen binding domain targeting a tumor antigen. In some embodiments, the induced pluripotent stem cell (iPSC) or a derivative cell thereof can further comprise at least one of (i) a deletion or reduced expression of one or more of B2M, TAP 1, TAP 2, Tapasin, RFXANK, CIITA, RFX5, RFXAP genes, (ii) an exogenous polynucleotide encoding a human leukocyte antigen E (HLA-E) and / or human leukocyte antigen G (HLA-G), (iii) an exogenous polynucleotide encoding a natural killer (NK) cell receptor immunoglobulin gamma Fc region receptor III (FcyRIII, cluster of differentiation 16 (CD16)) and / or an NKG2D protein, (iv) a deletion or reduced expression of one or both of NKG2A and CD70 genes, (v) an exogeneous polynucleotide encoding a cytokine, (vi) an exogenous polynucleotide encoding a safety switch, and optionally an exogeneous polynucleotide encoding a PSMA cell tracer. In some embodiments, the GPCR scaffold or fragment thereof is selected from the group consisting of rhodopsin-like receptors (Class A), secretin family receptors (Class B), or Metabotropic receptors (Class C). In some embodiments, the GPCR scaffold is a rhodopsin-like receptor (such as chemokine receptor). In some embodiments, the GPCR scaffold is selected from the group consisting of CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CCR10, CXCR1, Docket No.: 066461.11126 / 25WO1 CXCR2, CXCR3, CXCR4, CXCR5, CXCR6, CX3CR1, XCR1, ACKR1, ACKR2, ACKR3, ACKR4, and a fragment of any receptor thereof. In some embodiments of the present disclosure, the GPCR scaffold is CXCR3 or a fragment thereof. I. Induced Pluripotent Stem Cells (IPSCs) And Immune Effector Cells IPSCs have unlimited self-renewing capacity. Use of iPSCs enables cellular engineering to produce a controlled cell bank of modified cells that can be expanded and differentiated into desired immune effector cells, supplying large amounts of homogeneous allogeneic therapeutic products. Provided herein are genetically engineered IPSCs and derivative cells thereof. The selected genomic modifications provided herein enhance the therapeutic properties of the derivative cells. The derivative cells are functionally improved and suitable for allogenic off-the-shelf cell therapies following a combination of selective modalities being introduced to the cells at the level of iPSC through genomic engineering. This approach can help to reduce the side effects mediated by CRS / GVHD and prevent long- term autoimmunity while providing excellent efficacy. As used herein, the term "differentiation" is the process by which an unspecialized ("uncommitted") or less specialized cell acquires the features of a specialized cell. Specialized cells include, for example, a blood cell or a muscle cell. A differentiated or differentiation- induced cell is one that has taken on a more specialized ("committed") position within the lineage of a cell. The term "committed", when applied to the process of differentiation, refers to a cell that has proceeded in the differentiation pathway to a point where, under normal circumstances, it will continue to differentiate into a specific cell type or subset of cell types, and cannot, under normal circumstances, differentiate into a different cell type or revert to a less differentiated cell type. As used herein, the term "pluripotent" refers to the ability of a cell to form all lineages of the body or soma or the embryo proper. For example, embryonic stem cells are a type of pluripotent stem cells that are able to form cells from each of the three germs layers, the ectoderm, the mesoderm, and the endoderm. Pluripotency is a continuum of developmental potencies ranging from the incompletely or partially pluripotent cell (e.g., an epiblast stem cell or EpiSC), which is unable to give rise to a complete organism to the Docket No.: 066461.11126 / 25WO1 more primitive, more pluripotent cell, which is able to give rise to a complete organism (e.g., an embryonic stem cell). As used herein, the terms "reprogramming" or "dedifferentiation" refers to a method of increasing the potency of a cell or dedifferentiating the cell to a less differentiated state. For example, a cell that has an increased cell potency has more developmental plasticity (i.e., can differentiate into more cell types) compared to the same cell in the non-reprogrammed state. In other words, a reprogrammed cell is one that is in a less differentiated state than the same cell in a non-reprogrammed state. As used herein, the term "induced pluripotent stem cells" or, iPSCs, means that the stem cells are produced from differentiated adult, neonatal or fetal cells that have been induced or changed or reprogrammed into cells capable of differentiating into tissues of all three germ or dermal layers: mesoderm, endoderm, and ectoderm. The iPSCs produced do not refer to cells as they are found in nature. The term “hematopoietic stem and progenitor cells,” “hematopoietic stem cells,” “hematopoietic progenitor cells,” or “hematopoietic precursor cells” or “HPCs” refers to cells which are committed to a hematopoietic lineage but are capable of further hematopoietic differentiation. Hematopoietic stem cells include, for example, multipotent hematopoietic stem cells (hematoblasts), myeloid progenitors, megakaryocyte progenitors, erythrocyte progenitors, and lymphoid progenitors. Hematopoietic stem and progenitor cells (HSCs) are multipotent stem cells that give rise to all the blood cell types including myeloid (monocytes and macrophages, neutrophils, basophils, eosinophils, erythrocytes, megakaryocytes / platelets, dendritic cells), and lymphoid lineages (T cells, B cells, NK cells). As used herein, “CD34+ hematopoietic progenitor cell” refers to an HPC that expresses CD34 on its surface. As used herein, the term “immune cell” or “immune effector cell” refers to a cell that is involved in an immune response. Immune response includes, for example, the promotion of an immune effector response. Examples of immune cells include T cells, B cells, natural killer (NK) cells, mast cells, and myeloid-derived phagocytes. As used herein, the terms “T lymphocyte” and “T cell” are used interchangeably and refer to a type of white blood cell that completes maturation in the thymus and that has various roles in the immune system. A T cell can have the roles including, e.g., the Docket No.: 066461.11126 / 25WO1 identification of specific foreign antigens in the body and the activation and deactivation of other immune cells. A T cell can be any T cell, such as a cultured T cell, e.g., a primary T cell, or a T cell from a cultured T cell line, e.g., Jurkat, SupTl, etc., or a T cell obtained from a mammal. The T cell can be CD3+ cells. The T cell can be any type of T cell and can be of any developmental stage, including but not limited to, CD4+ / CD8+ double positive T cells, CD4+ helper T cells (e.g., Thl and Th2 cells), CD8+ T cells (e.g., cytotoxic T cells), peripheral blood mononuclear cells (PBMCs), peripheral blood leukocytes (PBLs), tumor infiltrating lymphocytes (TILs), memory T cells, naive T cells, regulator T cells, gamma delta T cells (gd T cells), and the like. Additional types of helper T cells include cells such as Th3 (Treg), Thl7, Th9, or Tfh cells. Additional types of memory T cells include cells such as central memory T cells (Tcm cells), effector memory T cells (Tern cells and TEMRA cells). The T cell can also refer to a genetically engineered T cell, such as a T cell modified to express a T cell receptor (TCR) or a chimeric antigen receptor (CAR). The T cell can also be differentiated from a stem cell or progenitor cell. “CD4+ T cells” refers to a subset of T cells that express CD4 on their surface and are associated with cell-mediated immune response. They are characterized by the secretion profiles following stimulation, which may include secretion of cytokines such as IFN-gamma, TNF-alpha, IL2, IL4 and IL10. “CD4” are 55-kD glycoproteins originally defined as differentiation antigens on T-lymphocytes, but also found on other cells including monocytes / macrophages. CD4 antigens are members of the immunoglobulin supergene family and are implicated as associative recognition elements in MHC (major histocompatibility complex) class II-restricted immune responses. On T- lymphocytes they define the helper / inducer subset. “CD8+ T cells” refers to a subset of T cells which express CD8 on their surface, are MHC class I-restricted, and function as cytotoxic T cells. “CD8” molecules are differentiation antigens found on thymocytes and on cytotoxic and suppressor T- lymphocytes. CD8 antigens are members of the immunoglobulin supergene family and are associative recognition elements in major histocompatibility complex class I- restricted interactions. Docket No.: 066461.11126 / 25WO1 As used herein, the term “NK cell” or “Natural Killer cell” refers to a subset of peripheral blood lymphocytes defined by the expression of CD56 and CD45 and the absence of the T cell receptor (TCR chains). The NK cell can also refer to a genetically engineered NK cell, such as a NK cell modified to express a chimeric antigen receptor (CAR). The NK cell can also be differentiated from a stem cell or progenitor cell. The induced pluripotent stem cell (iPSC) parental cell lines may be generated from peripheral blood mononuclear cells (PBMCs) or T-cells using any known method for introducing re-programming factors into non-pluripotent cells such as the episomal plasmid-based process as previously described in U.S. Pat. Nos.8,546,140; 9,644,184; 9,328,332; and 8,765,470, the complete disclosures of which are incorporated herein by reference. The reprogramming factors may be in a form of polynucleotides, and thus are introduced to the non-pluripotent cells by vectors such as a retrovirus, a Sendai virus, an adenovirus, an episome, and a mini-circle. In particular embodiments, the one or more polynucleotides encoding at least one reprogramming factor are introduced by a lentiviral vector. In some embodiments, the one or more polynucleotides introduced by an episomal vector. In various other embodiments, the one or more polynucleotides are introduced by a Sendai viral vector. In some embodiments, the iPSC’s are clonal iPSC’s or are obtained from a pool of iPSCs and the genome edits are introduced by making one or more targeted integration and / or in / del at one or more selected sites. In another embodiment, the iPSC’s are obtained from human T cells having antigen specificity and a reconstituted TCR gene (hereinafter, also refer to as "T-iPS” cells) as described in US Pat. Nos.9206394, and 10787642 hereby incorporated by reference into the present application. According to a particular aspect, the application relates to an induced pluripotent stem cell (iPSC) cell or a derivative cell thereof comprising: (i) an exogenous polynucleotide encoding a chimeric antigen receptor (CAR) comprising a G protein- coupled receptor (GPCR) scaffold or fragment thereof and at least one antigen binding domain targeting a tumor antigen; (ii) an exogenous polynucleotide encoding a truncated epithelial growth factor receptor (tEGFR) variant and an interleukin 15 (IL-15), wherein the tEGFR variant and IL-15 are operably linked by an autoprotease peptide sequence, Docket No.: 066461.11126 / 25WO1 such as the porcine tesehovirus-12A (P2A); and (iii) a deletion or reduced expression of B2M and CIITA genes. II. Expression of Chimeric Antigen Receptors Utilizing G Protein-coupled Receptor Scaffolds In some embodiments, an iPSC cell or derivative cell thereof contains an exogenous polynucleotide encoding a chimeric antigen receptor (CAR), such as a CAR targeting a tumor antigen, and said CAR comprises a GPCR scaffold. In some instances, the recombinant CAR polypeptide includes a GPCR scaffold comprising a plurality of transmembrane domains connected by intracellular and / or extracellular loops. In some embodiments, the recombinant CAR polypeptide further comprises at least one extracellular domain (disposed at an extracellular loop or an N-terminal tail of the GPCR scaffold) that binds specifically to an antigen (or more than one antigen). In some embodiments, the recombinant CAR polypeptide further comprises an intracellular signaling domain (disposed at an intracellular loop or C-terminal tail of the GPCR scaffold). In some instances, the recombinant CAR polypeptide includes a signal peptide, one or more intracellular signaling domains, and / or one or more co-stimulatory domains. A. G Protein-Coupled Receptor (GPCR) Scaffolds In certain embodiments, a CAR of the present disclosure comprises a GPCR scaffold. GPCRs are a large family of membrane receptors that detect molecules outside the cell and activate internal signal transduction pathways and cellular responses. Structurally, GPCRs typically contain multiple transmembrane domains, intracellular and extracellular loops, an extracellular N-terminal region, and an intracellular C-terminal tail. By engineering (i) intracellular domains on different intracellular loops (or the C terminus), and / or (ii) antigen binders on the N terminus or extracellular loops, multiple functional extracellular and intracellular domains can be engineered into a single CAR. The modularity, tuning capacity and signaling diversity of GPCRs make them promising scaffolds for engineering CARs with tailored signaling properties and potentially improved therapeutic responses as compared to standard CAR designs. GPCRs have a conserved structure with an extracellular ligand binding domain that can be engineered to bind specific antigens. Additionally, GPCRs couple to different G Docket No.: 066461.11126 / 25WO1 proteins which activate various effector proteins and cellular responses (e.g., proliferation, cytokine production), can have lower T cell activation thresholds, resulting in specific discrimination between target and healthy cells. In certain embodiments, a CAR of the present disclosure comprises a GPCR scaffold. Generally, scaffold proteins provide a platform to bring together multiple proteins involved in a particular signaling pathway. By interacting with various components, scaffold proteins facilitate signaling between membrane receptors and intracellular effectors. They coordinate multi-protein signaling complexes and ensure the specificity and efficiency of signal transduction. In other words, scaffolds can provide spatial and temporal control over signaling. In the case of CARs expressed on the surface of effector cells (e.g., T or NK cells), scaffold proteins (such as a GPCR) can be used, for example, to facilitate (i) the proximate surface expression of multiple tumor antigen binding domains (e.g., to enhance anti-tumor specificity) and / or (ii) intracellular signaling in response to successful tumor antigen binding by the effector cell. G protein-coupled receptors (GPCRs) (also known as seven-(pass)- transmembrane domain receptors, 7TM receptors, heptahelical receptors, serpentine receptors, and G protein-linked receptors (GPLR)) form a large group of evolutionarily related cell surface receptors that detect molecules outside the cell and activate cellular responses. They pass through the cell membrane seven times in form of six loops (i.e., three extracellular loops interacting with ligand molecules, three intracellular loops interacting with G proteins, an N-terminal extracellular region and a C-terminal intracellular region) of amino acid residues. According to the classical A-F system, GPCRs can be grouped into 6 classes based on sequence homology and functional similarity. More recently, an alternative classification system called GRAFS (Glutamate, Rhodopsin, Adhesion, Frizzled / Taste2, Secretin) has been proposed for vertebrate GPCRs. A person of skill in the art will appreciate that any GPCR may be used with embodiments of the present disclosure. In some embodiments of the present disclosure, the GPCR scaffold or fragment thereof is selected from the group consisting of rhodopsin-like receptors (Class A), secretin family receptors (Class B), and Metabotropic receptors (Class C). In certain embodiments, a GPCR comprising a native ligand binding domain may be modified to remove said ligand Docket No.: 066461.11126 / 25WO1 binding domain and replace it with an alternative ligand binding domain (e.g., a tumor antigen binding domain). Class A GPCRs, also known as rhodopsin-like receptors, make up the largest class of G protein-coupled receptors. They are characterized by a common structure consisting of a single polypeptide chain that weaves back and forth across the plasma membrane 7 times, forming 7 alpha helical transmembrane domains. The N-terminus of the receptor sits outside the cell, while the C-terminus resides inside the cell. The transmembrane helices are connected by 3 extracellular loops and 3 intracellular loops. Class A GPCRs bind to ligands such as hormones, neurotransmitters, cytokines and chemokines. The binding of these small diffusible ligands induces a conformational change in the transmembrane domains, activating the receptor. This causes the receptor to couple to a heterotrimeric G protein inside the cell, made up of alpha, beta and gamma subunits. Activation of the G protein leads to downstream signaling cascades that generate cellular responses. There are over 700 different Class A GPCRs known, making this by far the largest and most diverse group. Some examples of Class A GPCRs include rhodopsin, adrenergic, dopamine, serotonin and chemokine receptors. Class A receptors exhibit some conserved sequence motifs like the DRY motif, and their activity is regulated by phosphorylation, palmitoylation and other post-translational modifications. In some embodiments, the GPCR scaffold is a Class A GPCR. In some embodiments, the GPCR scaffold is a Class A GPCR selected from the group consisting of ACKR1, ACKR2, ACKR3, ACKR4, Blue opsin, CCR1, CCR10, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CNR1, CNR2, CX3CR1, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CXCR6, Encephalopsin / panopsin, GNRHR, GNRHR2, GPR1, GPR101, GPR119, GPR12, GPR132, GPR135, GPR139, GPR141, GPR142, GPR146, GPR148, GPR149, GPR15, GPR150, GPR151, GPR152, GPR153, GPR160, GPR161, GPR162, GPR17, GPR171, GPR173, GPR174, GPR176, GPR18, GPR182, GPR183, GPR20, GPR22, GPR25, GPR26, GPR27, GPR3, GPR31, GPR32, GPR33, GPR34, GPR35, GPR37, GPR39, GPR4, GPR45, GPR50, GPR52, GPR55, GPR6, GPR61, GPR62, GPR63, GPR65, GPR68, GPR75, GPR78, GPR79, GPR82, GPR83, GPR84, GPR85, GPR87, GPR88, GPRC5A, GPRC5B, GPRC5C, GPRC5D, Green opsin, LPAR1, LPAR2, LPAR3, LPAR4, LPAR5, LPAR6, MAS1, MC1R, MC2R, Docket No.: 066461.11126 / 25WO1 MC3R, MC4R, MC5R, Melanopsin, MRGE, Neuropsin (opn5), P2Y1, P2Y11, P2Y12, P2Y13, P2Y14, P2Y2, P2Y4, P2Y6, Peropsin, PTAFR, Red opsin, Retinal G protein coupled receptor, Rhodopsin, Rhodopsin 3, Rhodopsin 4, Rhodopsin 5, Rhodopsin 6, Rhodopsin 7, S1PR1, S1PR2, S1PR3, S1PR4, S1PR5, Teleost multiple tissue (tmt) opsin, Vertebrate ancient (VA) opsin, Vertebrate ancient 2 (VA2) opsin, Vertebrate ancient long (VAL) opsin, and XCR1. In certain embodiments, the GPCR scaffold is a chemokine receptor. In certain embodiments the chemokine receptor is selected from the group consisting of CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CCR10, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CXCR6, CX3CR1, XCR1, ACKR1, ACKR2, ACKR3, ACKR4, and a fragment of any receptor thereof. In some embodiments, the GPCR scaffold is CXCR3 or a fragment thereof. In some embodiments, the GPCR scaffold is CXCR3 or a fragment thereof, and (i) the CXCR3 comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 134, or (ii) the CXCR3 is encoded by a polynucleotide having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one or more of SEQ ID NOs: 135-151. Class B GPCRs, also known as secretin receptor family, represent a smaller group of around 15 receptors that bind peptide hormones. The peptides that activate Class B receptors are larger than Class A ligands. Examples include secretin, glucagon, calcitonin and other hormone peptides. Despite the differences in their extracellular regions, Class B receptors have a similar 7-transmembrane alpha helical structure spanning the membrane as Class A. One of the hallmarks of Class B receptors is their mechanism of ligand binding and activation. Their hormones bind to the extracellular N-terminal region, triggering a conformational change that propagates through the transmembrane domains. This induces movement of the ends of the transmembrane helices and subsequent G protein activation on the intracellular side. In some embodiments, the GPCR scaffold is a Class B GPCR. In some embodiments, the GPCR scaffold is a Class B GPCR selected from the group consisting of ADGRB1, ADGRB2, ADGRB3, ADGRD1, ADGRD2, ADGRE1, ADGRE2, ADGRE3, ADGRE4P, ADGRE5, ADGRF1, ADGRF2, ADGRF3, ADGRF4, ADGRF5, ADGRG1, ADGRG2, ADGRG3, ADGRG4, ADGRG5, ADGRG6, ADGRG7, a Docket No.: 066461.11126 / 25WO1 Calcitonin receptor, a CRH receptor, CRHR1, CRHR2, an FSH receptor, a GHRH receptor, a GIP receptor, a GLP-1 receptor, a GLP-2 receptor, a Glucagon receptor, a Glucagon-like peptide-1 receptor, a Glucagon-like peptide-2 receptor, an LH receptor, a PAC1 receptor, a PACAP receptor, a PTH1 receptor, a PTH2 receptor, a Secretin receptor, a TSH receptor, a VPAC1 receptor, and a VPAC2 receptor. Class C GPCRs, also known as metabotropic glutamate, pheromone and calcium- sensing receptors, are characterized by a large bilobed extracellular domain that serves as the ligand binding site. There are around 22 Class C receptors that can be activated by glutamate, pheromones, calcium ions and other ligands. Similar to Class B receptors, the extracellular domain of Class C receptors is responsible for ligand recognition and inducing receptor activation through conformational changes that get propagated to the transmembrane domains. However, the extracellular domain has a bipartite structure formed by two lobes connected by a hinge region, unlike the singular extracellular domain of Class B. Binding of ligands like glutamate and calcium to the cleft between the two lobes causes the lobes to come together. This closure exerts an effect on the heptahelical transmembrane domain to activate downstream signaling. Despite some differences, Class C receptors have 7 alpha helical transmembrane domains and couple to G proteins like Class A and B. Examples of Class C receptors include metabotropic glutamate receptors involved in neurotransmission, calcium-sensing receptors. In some embodiments, the GPCR scaffold is a Class C GPCR. In some embodiments, the GPCR scaffold is a Class C GPCR selected from the group consisting of CaSR, mGluR1, mGluR2, mGluR3, mGluR4, mGluR5, mGluR6, mGluR7, mGluR8, TAS1R1, TAS1R2, and TAS1R3. In some aspects, an iPSC or the derivative cell of the present disclosure can comprise a CAR having a GPCR scaffold. In some embodiments, the GPCR scaffold can comprise (i) at least one an extracellular domain comprising the first antigen-binding domain, (ii) at least one transmembrane domain; and / or (iii) at least one intracellular domain. B. Extracellular domains Docket No.: 066461.11126 / 25WO1 In certain embodiments, an extracellular domain of a GPCR CAR includes an antibody, an antibody fragment, an antigen-binding domain and / or an antigen-binding fragment. The antigen binding fragment can, for example, be an antibody or antigen- binding fragment thereof that specifically binds a tumor antigen. In some embodiments, the antigen-binding domains or fragments possess one or more desirable functional properties including, but not limited to, high-affinity binding to a tumor antigen, high specificity to a tumor antigen, the ability to stimulate complement-dependent cytotoxicity (CDC), antibody-dependent phagocytosis (ADPC), and / or antibody-dependent cellular- mediated cytotoxicity (ADCC) against cells expressing a tumor antigen, and the ability to inhibit tumor growth in subjects in need thereof and in animal models when administered alone or in combination with other anti-cancer therapies. In some embodiments, antibodies or antibody fragments suitable for use in the CAR include, but are not limited to, monoclonal antibodies, bispecific antibodies, multispecific antibodies, chimeric antibodies, polypeptide-Fc fusions, single-chain Fvs (scFv), single chain antibodies, Fab fragments, F(ab′) fragments, disulfide-linked Fvs (sdFv), masked antibodies (e.g., Probodies®), Small Modular ImmunoPharmaceuticals ("SMIPsTM"), intrabodies, minibodies, single domain antibody variable domains, nanobodies, VHHs, diabodies, tandem diabodies (TandAb®), anti-idiotypic (anti-Id) antibodies (including, e.g., anti-Id antibodies to antigen-specific TCR), and epitope- binding fragments of any of the above. Antibodies and / or antibody fragments may be derived from murine antibodies, rabbit antibodies, human antibodies, fully humanized antibodies, camelid antibody variable domains and humanized versions, shark antibody variable domains and humanized versions, and camelized antibody variable domains. In some embodiments, the antigen-binding fragment is an Fab fragment, an Fab' fragment, an F(ab')2 fragment, an scFv fragment, an Fv fragment, a dsFv diabody, a VHH, a VNAR, a single-domain antibody (sdAb) or nanobody, a dAb fragment, a Fd' fragment, a Fd fragment, a heavy chain variable region, an isolated complementarity determining region (CDR), a diabody, a triabody, or a decabody. In some embodiments, the antigen-binding fragment is an scFv fragment. In some embodiments, the antigen- binding fragment is a VHH. Docket No.: 066461.11126 / 25WO1 In some embodiments, the extracellular domain of the CAR is a single-domain antibody or nanobody. In some embodiments, the extracellular domain is a VHH. In some embodiments, the extracellular domain is an scFv. i. Antigen-binding domains Generally, a GPCR CAR of the present disclosure can comprise an extracellular domain comprising an antigen-binding domain. Such extracellular domains, for example, can consist of an extracellular loop (ECL) of the GPCR in which the antigen-binding domain is disposed. By expressing the antigen binding domain in an ECL of the GPCR scaffold, the antigen-binding domain can be expressed on the surface of the cell, and in proximity of other antigen binding domains or costimulatory / signalling domains expressed in another ECL domain of the same GPCR. In some embodiments, an antigen-binding domain of a CAR binds to a target antigen. The antigen-binding domain may bind to more than one antigen or more than one epitope in an antigen. For example, the antigen-binding domain may bind to 2, 3, 4, 5, 6, 7, 8 or more antigens. As another example, the antigen-binding domain may bind 2, 3, 4, 5, 6, 7, 8 or more epitopes in the same antigen. The choice of antigen-binding domain may depend upon the type and number of antigens that define the surface of a target cell. For example, the antigen-binding domain may be chosen to recognize an antigen that acts as a cell surface marker on target cells associated with a particular disease state. In some embodiments, CAR can be genetically modified to target a tumor antigen of interest by way of engineering a desired antigen- binding domain that specifically binds to an antigen (e.g., on a tumor cell). Non-limiting examples of cell surface markers that may act as targets for the antigen-binding domain in the CAR include those associated with tumor cells or autoimmune diseases. In some embodiments, the antigen-binding domain binds to at least one tumor antigen or autoimmune antigen. In some embodiments, the antigen-binding domain binds to at least one tumor antigen or autoimmune antigen selected from one or more of Tables 1-3. Docket No.: 066461.11126 / 25WO1 In some embodiments, the antigen-binding domain binds to at least one tumor antigen. In some embodiments, the antigen-binding domain binds to two or more tumor antigens. In some embodiments, the two or more tumor antigens are associated with the same tumor. In some embodiments, the two or more tumor antigens are associated with different tumors. In some embodiments, the antigen-binding domain binds to at least one autoimmune antigen. In some embodiments, the antigen-binding domain binds to two or more autoimmune antigens. In some embodiments, the two or more autoimmune antigens are associated with the same autoimmune disease. In some embodiments, the two or more autoimmune antigens are associated with different autoimmune diseases. In some embodiments, the tumor antigen is associated with glioblastoma, ovarian cancer, cervical cancer, head and neck cancer, liver cancer, prostate cancer, pancreatic cancer, renal cell carcinoma, bladder cancer, or hematologic malignancy. Non-limiting examples of tumor antigens associated with glioblastoma include HER2, EGFRvIII, EGFR, CD133, PDGFRA, FGFR1, FGFR3, MET, CD70, ROBO1 and IL13Rα2. Non- limiting examples of tumor antigens associated with ovarian cancer include FOLR1, FSHR, MUC16, MUC1, Mesothelin, CA125, EpCAM, EGFR, PDGFRα, Nectin-4 and B7H4. Non-limiting examples of the tumor antigens associated with cervical cancer or head and neck cancer include GD2, MUC1, Mesothelin, HER2, and EGFR. Non-limiting examples of tumor antigen associated with liver cancer include Claudin 18.2, GPC-3, EpCAM, cMET, and AFP. Non-limiting examples of tumor antigens associated with hematological malignancies include CD19, CD22, CD79, BCMA, GPRC5D, SLAM F7, CD33, CLL1, CD123, and CD70. Non-limiting examples of tumor antigens associated with bladder cancer include Nectin-4 and SLITRK6. Non-limiting examples of tumor antigens associated with renal cancer include CD70 and FOLR1. Additional examples of antigens that may be targeted by the antigen-binding domain include, but are not limited to, alpha-fetoprotein, A3, antigen specific for A33 antibody, Ba 733, BrE3-antigen, carbonic anhydrase EX, CD1, CD1a, CD3, CD5, CD15, CD16, CD19, CD20, CD21, CD22, CD23, CD25, CD30, CD33, CD38, CD45, CD74, CD79a, CD80, CD123, CD138, colon-specific antigen-p (CSAp), CEA (CEACAM5), Docket No.: 066461.11126 / 25WO1 CEACAM6, CSAp, EGFR, EGP-I, EGP-2, Ep-CAM, EphA1, EphA2, EphA3, EphA4, EphA5, EphA6, EphA7, EphA8, EphA10, EphB1, EphB2, EphB3, EphB4, EphB6, FIt-I, Flt-3, folate receptor, HLA-DR, human chorionic gonadotropin (HCG) and its subunits, hypoxia inducible factor (HIF-I), Ia, IL-2, IL-6, IL-8, insulin growth factor-1 (IGF-I), KC4-antigen, KS-1-antigen, KS1-4, Le-Y, macrophage inhibition factor (MIF), MAGE, MUC2, MUC3, MUC4, NCA66, NCA95, NCA90, antigen specific for PAM-4 antibody, placental growth factor, p53, prostatic acid phosphatase, PSA, PSMA, RS5, S100, TAC, TAG-72, tenascin, TRAIL receptors, Tn antigen, Thomson-Friedenreich antigens, tumor necrosis antigens, VEGF, ED-B fibronectin, 17-1A-antigen, an angiogenesis marker, an oncogene marker or an oncogene product. In some embodiments, the antigen is associated with an autoimmune disease or disorder. Such antigens may be derived from cell receptors and cells which produce “self”-directed antibodies. In some embodiments, the antigen is associated with an autoimmune disease or disorder such as Rheumatoid arthritis (RA), multiple sclerosis (MS), Sjögren's syndrome, Systemic lupus erythematosus, sarcoidosis, type 1 diabetes mellitus, insulin dependent diabetes mellitus (IDDM), autoimmune thyroiditis, reactive arthritis, ankylosing spondylitis, scleroderma, polymyositis, dermatomyositis, psoriasis, vasculitis, Wegener's granulomatosis, Myasthenia gravis, Hashimoto's thyroiditis, Graves' disease, chronic inflammatory demyelinating polyneuropathy, Guillain-Barre syndrome, Crohn's disease or ulcerative colitis. In some embodiments, autoimmune antigens that may be targeted by the CAR include, but are not limited to, platelet antigens, myelin protein antigen, Sm antigens in snRNPs, islet cell antigen, rheumatoid factor, and anticitrullinated protein. citrullinated proteins and peptides such as CCP-1, CCP-2 (cyclical citrullinated peptides), fibrinogen, fibrin, vimentin, filaggrin, collagen I and II peptides, alpha-enolase, translation initiation factor 4G1, perinuclear factor, keratin, Sa (cytoskeletal protein vimentin), components of articular cartilage such as collagen II, IX, and XI, circulating serum proteins such as RFs (IgG, IgM), fibrinogen, plasminogen, ferritin, nuclear components such as RA33 / hnRNP A2, Sm, eukaryotic translation elongation factor 1 alpha 1, stress proteins such as HSP- 65, -70, -90, BiP, inflammatory / immune factors such as B7-H1, IL-1 alpha, and IL-8, enzymes such as calpastatin, alpha-enolase, aldolase-A, dipeptidyl peptidase, Docket No.: 066461.11126 / 25WO1 osteopontin, glucose-6-phosphate isomerase, receptors such as lipocortin 1, neutrophil nuclear proteins such as lactoferrin and 25-35 kD nuclear protein, granular proteins such as bactericidal permeability increasing protein (BPI), elastase, cathepsin G, myeloperoxidase, proteinase 3, platelet antigens, myelin protein antigen, islet cell antigen, rheumatoid factor, histones, ribosomal P proteins, cardiolipin, vimentin, nucleic acids such as dsDNA, ssDNA, and RNA, ribonuclear particles and proteins such as Sm antigens (including but not limited to SmD's and SmB′ / B), U1RNP, A2 / B1 hnRNP, Ro (SSA), and La (SSB) antigens. Non-limiting exemplary antigen targets are provided in Tables 1-3. Table 1 provides antigen binding domains that bind to exemplary antigen targets. The antigen- binding domain may comprise a VH sequence, a VL sequence, and / or CDRs thereof, such as those described in the cited publications, the contents of each publication are incorporated herein by reference in their entirety for all purposes. Table 1. Antigen target Type Examples of Source Type Examples of Source ; 4, Docket No.: 066461.11126 / 25WO1 APCDD1 VH Identifier 10, 102, 106, VL Identifier 136, 110, 114, 118, 122, 100, 104, 108, 12 1 1 4 14 112 11 12 120, 6, n 1, 9, ; ; Docket No.: 066461.11126 / 25WO1 , 280, 286, 292, 185, 191, 197, , 304, 310, 316, 203, 209, 215, 2 4 4 221 2272 , 0, 5, ; n , , 6, , ; ; ; Docket No.: 066461.11126 / 25WO1 BMPR1A VH Identifier 12 in - - WO2011116212 2, ; ; ; Docket No.: 066461.11126 / 25WO1 US20140134142 A1; Identifier 54, i ; 1, ; ; 3 ; 2 4 , Docket No.: 066461.11126 / 25WO1 US20160319020; Identifier 17, 33, 34, 35 i EP 7 4A1 ; ; ; ; ; ; ; ; ; Docket No.: 066461.11126 / 25WO1 WO2016164731; Identifier 686 in Identifier 679 in WO2016164731; W 211471 I ifi 7 i ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; Docket No.: 066461.11126 / 25WO1 WO2016164731; Identifier 760 in Identifier 722 in WO2016164731; W 211471 I ifi 71 i ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; Docket No.: 066461.11126 / 25WO1 WO2014144722 A2; WO2016126213 Identifier 12 in A1 W 211221A1 2, 2, 2, 2, , 0, ; 2, n ; 7, 5, Docket No.: 066461.11126 / 25WO1 75, 79, 83, 69, 71, 83 in 69, 73, 77, 81, 85, WO2012031273; 17, 73, 77 in I ifi 11 1112 W 212 127; 9, 4, 9, n ; ; ; Docket No.: 066461.11126 / 25WO1 730, 731, 732, 733, 734, 735, 7 7 7 7 Docket No.: 066461.11126 / 25WO1 893, 894, 895, 896, 897, 898, 1 ; ; 2 4, 9, 4, 9 2, 1, 1, Docket No.: 066461.11126 / 25WO1 125, 126, 127, 128, 65, 69, 73, 77 in 129, 130, 131, 132, WO2016073649 1 1 4 1 1 A1 n 8, ; 8, 3, 8, ; ; Docket No.: 066461.11126 / 25WO1 205, 207, 209, 21, 211, 178, 180, 182, 213, 23, 25, 27, 29, 31, 184, 186, 190, 7 41 4 1 2 1 4 1 2, 4, 4, 6, 6, 6, 6, 6, 7, ; , ; ; 7, n ; Docket No.: 066461.11126 / 25WO1 Identifier 4 in Identifier 2 in US8697845; Identifier US8697845; 1 i 21 2 24 I ifi 2 i ; 2, 2, ; ; ; 3, 8, 3, ; ; ; Docket No.: 066461.11126 / 25WO1 Identifier 2123 in Identifier 4051, WO2018231759 4052, 4053, 4054, 4 4 4 7, 0, 3, 6, 9, 2, 5, 8, 1, 4, 7, 0, 3, 6, 9, , 5, 8, 1, 4, 7, 0, 3, 6, 9, 2, 5, 8, 1, 4, 7, 0, 3, 6, 9, 2, 5, 8, 1, 4, 7, 0, 3, 6, Docket No.: 066461.11126 / 25WO1 4187, 4188, 4189, 4190, 4191, 4192, 41 41 4 41 5, 8, 1, 4, 9, 2, 5, 8, 1, 4, 7, 0, 3, 6, 9, 2, 5, 8, 1, 4, 7, 0, 3, 6, 9, 2, 5, 8, 8, 1, 4, 7, 0, 3, 6, 9, 2, 5, 8, 1, Docket No.: 066461.11126 / 25WO1 4322, 4323, 4324, 4325, 4326, 4327, 4 2 4 2 4 0, 3, 6, 9, 2, 5, 8, 1, 4, 7, 0, 3, 6, 9, 2, 5, 8, 1, 4, 7, 0, 3, 6, 9, 2, 5, 8, 1, 4, 7, 0, 3, 6, 9, 2, 5, 8, 1, 4, 7, 0, 3, 6, 9, Docket No.: 066461.11126 / 25WO1 4460, 4461, 4462, 4463, 4464, 4465, 44 44 7 44 8, 1, 4, 7, 0, 3, 6, 9, 2, 5, 8, 1, 4, 7, 0, 3, 6, 9, 2, 5, 8, 1, 4, 7, 0, 3, 6, 9, 2, 5, 8, 1, 4, 7, 0, 3, 6, 9, 2, 5, 8, 1, 4, 7, Docket No.: 066461.11126 / 25WO1 4598, 4599, 4600, 4601, 4602, 4603, 4 4 4 4 6, 9, 2, 5, 8, 1, 4, 7, 0, 3, 6, 9 2, 5, 8, 1, 4, 7, ; ; ; , 8, 3, ; 4, Docket No.: 066461.11126 / 25WO1 EphA2 receptor VH Identifier 20, 22, 24, VL Identifier 26, 28, 32, 34, 36, 37, 38, 40, 30, 47, 48, 49, 50, 42 4 4 74 7 i 2 7 i ; 4, ; ; ; ; 5, Docket No.: 066461.11126 / 25WO1 Identifier 1 in US20130216528; US20130216528; Identifier 11, 12 I ifi 1 i i ; 4, 3, 8, , Docket No.: 066461.11126 / 25WO1 Identifier 3 in WO2016168769 A1 I ifi 9, 2 Docket No.: 066461.11126 / 25WO1 38, 40, 84, 86, 88 in 285, 287, 37, 39, US20160168242 41, 87 in 21 1 242 8 ; ; n 6, 6, 0, ; Docket No.: 066461.11126 / 25WO1 Identifier 1 in WO2015042246 4, ; 2 2, ; Docket No.: 066461.11126 / 25WO1 Identifier 5, 6 in WO2013142034; WO2013142034; Identifier 11, 15, I ifi 1 2 i 1 2 27 i ; n ; 5 Docket No.: 066461.11126 / 25WO1 MVR VH Identifier 1 in VL Identifier 5 in US20160257762A1 US20160257762 A1 n ; 8 n n Docket No.: 066461.11126 / 25WO1 28, 318, 37, 48, 50, 58, US9428570; 66,, 74, 85, 93, 95, 97, Identifier 116, i W 211 22 12 122 4, ; 5, ; 7, 7, ; 7, ; ; ; ; ; ; Docket No.: 066461.11126 / 25WO1 Identifier 10 in US20150190506 4, ; 9 ; ; 0, 8, 1 Docket No.: 066461.11126 / 25WO1 US20160333114 A1 ; 2, 2, 2, 2, , 2, 8 , ; ; 9, Docket No.: 066461.11126 / 25WO1 EP3083671A1; 167, 175, 183, Identifier 104, 112, 191, 199, 207, 12 12 1 2 1 1 21 22 2 2 1, 1, 5, 0, 5, Docket No.: 066461.11126 / 25WO1 77, 85, 93 in WO2016179319A1; I ifi 7 i , ; ; , Docket No.: 066461.11126 / 25WO1 163, 164, 165, 166, 167 in 21 427 6, 1, a e prov es exempary an gen arges. e an gen- n ng oman may comprise an scFv derived from an antibody or antibody fragment that binds to an antigen target such as those described in the cited publications, the contents of each publication are incorporated herein by reference in their entirety for all purposes. Table 2. Antigen Target Examples of Source Activated alpha- Identifier 824 in US20090117096A1 2, 1, 6, 3, Docket No.: 066461.11126 / 25WO1 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 251 in US20160311907A1 CCR4 Identifier 7 9 in WO2015191997 2, 7, , , n , 9 5, 4, 4, Docket No.: 066461.11126 / 25WO1 CD33 Identifier 262, 263, 264, 265, 266, 267, 268, 39, 40, 41, 42, 43, 44, 45, 46, 47 in WO2016014576; Identifier 37 in WO2015092024 A2; Identifier 37 in EP3083691A2; Identifier 153 154 155 156 157 158 159 160 161 ; 2, 3, 6, 2, 4, 6, 8, 8, 8, 8, 8, Docket No.: 066461.11126 / 25WO1 850, 852, 854, 856, 858, 860, 862, 864, 866, 868, 870, 874, 876, 878, 880, 882, 884, 886, 888, 890, 892, 894, 896, 650, 678 in WO2016090337 Folate rece tor Identifier 15 in US20170002072A1 2, , , n Docket No.: 066461.11126 / 25WO1 Ranibizumab Identifier 40 in US20160208021; Identifier 40 in WO2016112870 RAS Identifier 81 in WO2016154047 Rit ximb Idntifir 36 in US20160208021 Idntifir 36 in WO2016112870 a e prov es exempary an gen arges. e an gen- n ng oman may comprise an antigen-binding domain derived from a CAR that binds to an antigen target, such as those described in the cited publications, the contents of each publication are incorporated herein by reference in their entirety for all purposes. Table 3. Antigen Target Examples of Source Acid / base leucine Identifier 3435 in WO2016124930 , , , , , Docket No.: 066461.11126 / 25WO1 WO2016094304 A3; Identifier 4, 5, 6, 8, 9, 10, 11, 12 in WO2013154760; Identifier 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 27 28 29 71 73 in WO2016014789; Identifier 125 126 127 , , , , , , , , , , Docket No.: 066461.11126 / 25WO1 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160 172 173 174 175 176 177 178 179 180 181 182 183 184, 8, in , in , , , , Docket No.: 066461.11126 / 25WO1 CD30 Identifier 20 in WO2016008973A1; Identifier 1 in WO2016116035A1; WO2016134284 (no Identifier); Identifier 2 in WO2016008973 , 0, 1 r in 2 Docket No.: 066461.11126 / 25WO1 Fra Identifier 959 in WO2016090337; Identifier 13 in US20170002072A1 GCN4 Identifier 8 10 in US9446105B2 4; 4, Docket No.: 066461.11126 / 25WO1 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86 in US20160311917A1; Identifier 26, 39 in WO2016172537A1; Identifier 40 in US20160311907A1; Identifier , , , , r 6, ii. Linkers In some embodiments, an scFv fragment of an extracellular domain of a CAR includes a linker between the VH and VL domains. The linker can be a peptide linker and may include any naturally occurring amino acid. Exemplary amino acids that may be included into the linker are Gly, Ser Pro, Thr, Glu, Lys, Arg, Ile, Leu, His and Phe. The Docket No.: 066461.11126 / 25WO1 linker should have a length that is adequate to connect the VH and the VL in such a way that they form the correct conformation relative to one another so that they retain the desired activity, such as binding to an antigen. The linker may be about 5-50 amino acids long. In some embodiments, the linker is about 10-40 amino acids long. In some embodiments, the linker is about 10-35 amino acids long. In some embodiments, the linker is about 10-30 amino acids long. In some embodiments, the linker is about 10-25 amino acids long. In some embodiments, the linker is about 10-20 amino acids long. In some embodiments, the linker is about 15-20 amino acids long. Exemplary linkers that may be used are Gly rich linkers, Gly and Ser containing linkers, Gly and Ala containing linkers, Ala and Ser containing linkers, and other flexible linkers. In some embodiments, the linker is a Whitlow linker. In one embodiment, the Whitlow linker includes the amino acid sequence set forth in SEQ ID NO:3, or a variant thereof having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%, sequence identity to SEQ ID NO:3 of PCT / US2021 / 072646. In another embodiment, the linker is a (G4S)3linker. In one embodiment, the (G4S)3linker includes the amino acid sequence set forth in SEQ ID NO: 25, or a variant thereof having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%, sequence identity to SEQ ID NO:25 of PCT / US2021 / 072646. Other linker sequences may include portions of immunoglobulin hinge area, CL or CH1 derived from any immunoglobulin heavy or light chain isotype. Exemplary linkers that may be used include any of SEQ ID NOs:26-56 in Table 1 of PCT / US2021 / 072646, the disclosure including the sequence listing is incorporated herein by reference. Additional linkers are described for example in WO2019 / 060695, incorporated by reference herein in its entirety. The linkers described herein including SEQ ID NOS: 40-73 of Table 4 can be used in any of the polypeptides provided including those containing CD16, NKG2D, IL- 15, IL-15Ra, HLA-E, HLA-G, HSV-TK, PSMA, and the like. Docket No.: 066461.11126 / 25WO1 Table 4 provides exemplary linkers (SEQ ID NOS:40-73), which correspond to SEQ ID NOS: 3 and 25-56 of US Application No.17 / 657,803 filed April 4, 2022, the contents of which are incorporated herein by reference in its entirety. Table 4. Name Linker Sequence SEQ ID SEQ ID NO: NO: of 3 Docket No.: 066461.11126 / 25WO1 Linker 19 STAGDTHLGGEDFD 59 42 Li k 20 GEGGSGEGGSGEGGS 60 43 C. Signal peptides In some embodiments, a CAR polypeptide includes a signal peptide (e.g., a leader peptide or localization peptide). The signal peptide may be positioned at the N-terminus of the extracellular domain. The signal peptide may be optionally cleaved from the extracellular domain during cellular processing and localization of the CAR to the cellular membrane. Any of various signal peptide sequences known to one of skill in the art may be used. Non-limiting examples of signal peptides from which the sequence may be derived include granulocyte-macrophage colony-stimulating factor receptor (GMCSFR), FcεR, human immunoglobulin (IgG) heavy chain (HC) variable region, CD8α, or any of various other proteins secreted by T cells. In some embodiments, the Docket No.: 066461.11126 / 25WO1 signal sequence is compatible with the secretory pathway of a T cell. In certain embodiments, the signal sequence is derived from a human immunoglobulin heavy chain. In some embodiments, the signal sequence is derived from GMCSFR. In one embodiment, the GMCSFR signal sequence includes the amino acid sequence set forth in SEQ ID NO: 1, or a variant thereof having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%, sequence identity to SEQ ID NO:1 as set forth in PCT / US2021 / 072646, the disclosure of which is incorporated herein by reference. Table 11. CAR regions Sequence SEQ ID NO D. Hinge regions Docket No.: 066461.11126 / 25WO1 In some embodiments, a GPCR CAR polypeptide includes a hinge region (e.g., a spacer region) between an extracellular domain and a transmembrane domain, such that the extracellular domain, hinge region, and transmembrane domain are in frame with each other. In some embodiments, the hinge can comprise the flexible N-terminus of a GPCR scaffold (e.g., CXCR3). In other embodiments, the flexible N-terminus of the GPCR scaffold can be substituted with a hinge region from another protein (e.g., CD8). A hinge region may contain up to 300 amino acids, preferably 10 to 100 amino acids and most preferably 25 to 50 amino acids. A hinge region may be derived from all or part of naturally occurring molecules, such as from all or part of the extracellular region of CD8, CD4 or CD28, or from all or part of an antibody constant region. Alternatively, the hinge region may be a synthetic sequence that corresponds to a naturally occurring spacer region sequence, or may be an entirely synthetic spacer region sequence. Non-limiting examples of hinge regions include a part of human CD8α chain, partial extracellular domain of CD28, FcγRlIIIa receptor, IgG, IgM, IgA, IgD, IgE, an Ig hinge, or functional fragment thereof. In some embodiments, additional linking amino acids are added to the hinge region to ensure that the antigen-binding domain is an optimal distance from the transmembrane domain. In some embodiments, when the hinge region is derived from an immunoglobulin, the region may be mutated to prevent Fc receptor binding. In some embodiments, the hinge region includes a hinge domain of a recognized protein. The hinge domain may be derived from CD8α, CD28, or an immunoglobulin (IgG). For example, the IgG hinge may be from IgG1, IgG2, IgG3, IgG4, IgM1, IgM2, IgA1, IgA2, IgD, IgE, or a chimera thereof. In some embodiments, the hinge domain comprises an immunoglobulin IgG hinge or functional fragment thereof. In certain embodiments, the IgG hinge is from IgG1, IgG2, IgG3, IgG4, IgM1, IgM2, IgA1, IgA2, IgD, IgE, or a chimera thereof. In various embodiments, the hinge domain comprises the CH1, CH2, CH3 and / or hinge region of the immunoglobulin. In many embodiments, the hinge domain comprises the core hinge region of the immunoglobulin. The term “core hinge” can be used interchangeably with the term “short hinge” (“SH”). Non-limiting examples of suitable hinge domains are the Docket No.: 066461.11126 / 25WO1 core immunoglobulin hinge regions include EPKSCDKTHTCPPCP (SEQ ID NO:57 of PCT / US2021 / 072646) from IgG1, ERKCCVECPPCP (SEQ ID NO: 58 of PCT / US2021 / 072646) from IgG2, ELKTPLGDTTHTCPRCP(EPKSCDTPPPCPRCP)3(SEQ ID NO: 59 of PCT / US2021 / 072646) from IgG3, and ESKYGPPCPSCP (SEQ ID NO: 60 of PCT / US2021 / 072646) from IgG4 (see also Wypych et al., JBC 2008283(23): 16194-16205, which is incorporated herein by reference in its entirety for all purposes). In many embodiments, the hinge domain is a fragment of the immunoglobulin hinge. In some embodiments, the hinge domain is derived from CD8 or CD28. In one embodiment, the CD8 hinge domain comprises the amino acid sequence set forth in SEQ ID NO:21, or a variant thereof having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%, sequence identity to SEQ ID NO:21 of PCT / US2021 / 072646, the disclosure of which is incorporated herein by reference. In one embodiment, the CD28 hinge domain comprises the amino acid sequence set forth in SEQ ID NO:22, or a variant thereof having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%, sequence identity to SEQ ID NO:22 of PCT / US2021 / 072646, the disclosure of which is incorporated herein by reference. In some embodiments, the transmembrane domain and / or hinge domain is derived from CD8 or CD28. In some embodiments, both the transmembrane domain and hinge domain are derived from CD8. In some embodiments, both the transmembrane domain and hinge domain are derived from CD28. E. Cytoplasmic domains including co-stimulatory domains In some aspects, a CAR polypeptide includes a cytoplasmic or intracellular domain (e.g., an intracellular loop of the GPCR), which contains at least one intracellular signaling domain. In some embodiments, a cytoplasmic domain also comprises one or more co-stimulatory signaling domains. The cytoplasmic domain is responsible for activation of at least one of the normal effector functions (e.g., specialized function) of the host cell (e.g., T cell) in Docket No.: 066461.11126 / 25WO1 which the CAR has been placed in. The term “effector function” refers to a specialized function of a cell. Effector function of a T-cell, for example, may be cytolytic activity or helper activity including the secretion of cytokines. A signaling domain can include a portion of a protein which transduces the effector function signal and directs the cell to perform a specialized function. While usually the entire signaling domain is present, in many cases it is not necessary to use the entire domain. To the extent that a truncated portion of the intracellular signaling domain is used, such truncated portion may be used in place of the intact chain as long as it transduces the effector function signal. An intracellular signaling domain includes any truncated portion of the signaling domain sufficient to transduce the effector function signal. Non-limiting examples of signaling domains which can be used include, e.g., signaling domains derived from DAP10, DAP12, Fc epsilon receptor I γ chain (FCER1G), FcRβ, CD3δ, CD3ε, CD3γ, CD3ζ, CD5, CD22, CD226, CD66d, CD79A, and CD79B. In some embodiments, the cytoplasmic domain comprises a CD3ζ signaling domain. In some embodiments, the CD3ζ signaling domain includes the amino acid sequence set forth in SEQ ID NO:6, or a variant thereof having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%, sequence identity to SEQ ID NO:6 of PCT / US2021 / 072646, the disclosure of which is incorporated herein by reference in its entirety. In some embodiments, the cytoplasmic domain contains one or more co- stimulatory signaling domains. In some embodiments, the one or more co-stimulatory signaling domains are derived from CD28, 41BB, IL2Rb, CD40, OX40 (CD134), CD80, CD86, CD27, ICOS, NKG2D, DAP10, DAP12, 2B4 (CD244), BTLA, CD30, GITR, CD226, CD79A, and HVEM in its entirety. In some embodiments, the co-stimulatory signaling domain is derived from 4- 1BB. In one embodiment, the 4-1BB co-stimulatory signaling domain comprises the amino acid sequence set forth in SEQ ID NO:8, or a variant thereof having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%, Docket No.: 066461.11126 / 25WO1 sequence identity to SEQ ID NO:8 of PCT / US2021 / 072646, the disclosure of which is incorporated herein by reference in its entirety. In some embodiments, the co-stimulatory signaling domain is derived from IL2Rb . In one embodiment, the IL2Rb co-stimulatory signaling domain comprises the amino acid sequence set forth in SEQ ID NO:9, or a variant thereof having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%, sequence identity to SEQ ID NO:9 of PCT / US2021 / 072646, the disclosure of which is incorporated herein by reference in its entirety. In some embodiments, the co-stimulatory signaling domain is derived from CD40. In one embodiment, the CD40 co-stimulatory signaling domain includes the amino acid sequence set forth in SEQ ID NO:10, or a variant thereof having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%, sequence identity to SEQ ID NO:10 of PCT / US2021 / 072646, the disclosure of which is incorporated herein by reference in its entirety. In some embodiments, the co-stimulatory signaling domain is derived from OX40. In one embodiment, the OX40 co-stimulatory signaling domain includes the amino acid sequence set forth in SEQ ID NO:11, or a variant thereof having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%, sequence identity to SEQ ID NO:11 of PCT / US2021 / 072646, the disclosure of which is incorporated herein by reference in its entirety. In some embodiments, the co-stimulatory signaling domain is derived from CD80. In one embodiment, the CD80 co-stimulatory signaling domain includes the amino acid sequence set forth in SEQ ID NO:12, or a variant thereof having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%, sequence identity to SEQ ID NO:12 of PCT / US2021 / 072646, the disclosure of which is incorporated herein by reference in its entirety. Docket No.: 066461.11126 / 25WO1 In some embodiments, the co-stimulatory signaling domain is derived from CD86. In one embodiment, the CD86 co-stimulatory signaling domain includes the amino acid sequence set forth in SEQ ID NO: 13, or a variant thereof having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%, sequence identity to SEQ ID NO:13 of PCT / US2021 / 072646, the disclosure of which is incorporated herein by reference in its entirety. In some embodiments, the co-stimulatory signaling domain is derived from CD27. In one embodiment, the CD27 co-stimulatory signaling domain includes the amino acid sequence set forth in SEQ ID NO:14, or a variant thereof having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%, sequence identity to SEQ ID NO:14 of PCT / US2021 / 072646, the disclosure of which is incorporated herein by reference in its entirety. In some embodiments, the co-stimulatory signaling domain is derived from ICOS. In one embodiment, the ICOS co-stimulatory signaling domain includes the amino acid sequence set forth in SEQ ID NO:15, or a variant thereof having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%, sequence identity to SEQ ID NO:15 of PCT / US2021 / 072646, the disclosure of which is incorporated herein by reference in its entirety. In some embodiments, the co-stimulatory signaling domain is derived from NKG2D. In one embodiment, the NKG2D co-stimulatory signaling domain includes the amino acid sequence set forth in SEQ ID NO:16, or a variant thereof having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%, sequence identity to SEQ ID NO:16 of PCT / US2021 / 072646, the disclosure of which is incorporated herein by reference in its entirety. In some embodiments, the co-stimulatory signaling domain is derived from DAP10. In one embodiment, the DAP10 co-stimulatory signaling domain includes the Docket No.: 066461.11126 / 25WO1 amino acid sequence set forth in SEQ ID NO: 17, or a variant thereof having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%, sequence identity to SEQ ID NO:17 of PCT / US2021 / 072646, the disclosure of which is incorporated herein by reference in its entirety. In some embodiments, the co-stimulatory signaling domain is derived from DAP12. In one embodiment, the DAP12 co-stimulatory signaling domain includes the amino acid sequence set forth in SEQ ID NO:18, or a variant thereof having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%, sequence identity to SEQ ID NO:18 of PCT / US2021 / 072646, the disclosure of which is incorporated herein by reference in its entirety. In some embodiments, the co-stimulatory signaling domain is derived from 2B4 (CD244). In one embodiment, the 2B4 (CD244) co-stimulatory signaling domain includes the amino acid sequence set forth in SEQ ID NO:19, or a variant thereof having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%, sequence identity to SEQ ID NO:19 of PCT / US2021 / 072646, the disclosure of which is incorporated herein by reference in its entirety. In some embodiments, the CAR polypeptide includes one costimulatory signaling domains. In many embodiments, the CAR includes 2 or more costimulatory signaling domains. In various embodiments, the CAR includes 2, 3, 4, 5, 6, or more costimulatory signaling domains. In some embodiments, the signaling domain(s) and co-stimulatory signaling domain(s) can be placed in any order. In some embodiments, the signaling domain is upstream of the co-stimulatory signaling domains. In some embodiments, the signaling domain is downstream from the co-stimulatory signaling domains. In the cases where two or more co-stimulatory domains are included, the order of the co-stimulatory signaling domains could be switched. Docket No.: 066461.11126 / 25WO1 Non-limiting exemplary CAR polypeptide sequences and polynucleotide sequences, and portions thereof, are provided in Table 7, including sequences for various CAR constructs shown in Figure 1A-H.

[0002] Docket No.: 066461.11126 / 25WO1 Table 7. C CXC WT C RECE ACID WT C RECE ACID GTGGTTGTTGCTTTCGCCTTATGCTGGACCCCATACCATTTAGTTGTGCTGGTGGA

[0003] Docket No.: 066461.11126 / 25WO1 CXC TAIL SEQU CXC TAIL SEQU CXC AMI SEQU CXC NUC SEQU CXC INTR LOO ACID CXC INTRACELLULAR

[0004] Docket No.: 066461.11126 / 25WO1 LOO NUC SEQU CXC AMI SEQU CXC NUC SEQU CXC EXTR LOO ACID CXC EXTR LOO NUC SEQU CXC AMI SEQU

[0005] Docket No.: 066461.11126 / 25WO1 CXC NUC SEQU CXC ACID CXC NUC SEQU CXC AMI SEQU CXC NUC SEQU CXC ACID CXC NUC SEQU

[0006] Docket No.: 066461.11126 / 25WO1 CXC AMI SEQU CXC NUC SEQU CXC ACID CXC NUC SEQU CXC AMI SEQU CXC NUC SEQU CXC ACID

[0007] Docket No.: 066461.11126 / 25WO1 CXC NUC SEQU CXC AMI SEQU CXC NUC SEQU CXC AMI SEQU CXC NUC SEQU CXC TAIL SEQU CXC TAIL SEQU

[0008] Docket No.: 066461.11126 / 25WO1 CO-S 41BB (AA 2 41BB SEQU IL2R (AA 2 CD40 (AA 2 OX40 (AA 2 OX40 SEQU CD80 (AA 2 CD86 CD27 (AA 2

[0009] Docket No.: 066461.11126 / 25WO1 ICOS (AA 1 NKG (AA 1 DAP1 (AA 7 DAP1 (AA 6 2B4 / C 370) CD3- 3 ITA DOM (AA 5 CD3- 3 ITA DOM ACID

[0010] Docket No.: 066461.11126 / 25WO1 CD28 (AA 1 CD28 SEQU IL18R (373- IL18R DOM ANTI ANTI FMC FMC (NUC GACAACCTACTACAACAGCGCCCTGAAGTCCCGGCTGACCATCATCAAGGACAA

[0011] Docket No.: 066461.11126 / 25WO1 FMC ACID FMC (NUC CD22 ACID CD22 A01 ( CD22 A01 ( CCAAGAATACCTTGTACCTGCAAATGAATAGCCTTCGCGCAGAAGACACAGCGG

[0012] Docket No.: 066461.11126 / 25WO1 CD22 A01 ( CD22 A01 ( CD22 04 (A CD22 04 (N

[0013] Docket No.: 066461.11126 / 25WO1 DO4- (AMI DO4- (NUC PANI (AMI PANI (NUC PANI (AMI

[0014] Docket No.: 066461.11126 / 25WO1 PANI (NUC RW0 (AMI RW0 (NUC RW0 (AMI RW0 (NUC TCCAAAAACACAGCCTACCTACAAATGAACAGCTTAAGAGCTGAGGACACTGCC

[0015] Docket No.: 066461.11126 / 25WO1 DB01 P211 (AMI DB01 P210 (AMI DB01 P211 (AMI DB01 P212 (AMI DB01 P210 (AMI DB01 P212 (AMI

[0016] Docket No.: 066461.11126 / 25WO1 NEC_ ACID NEC_ ACID NEC_ P311 (AMI NEC_ ACID NEC_ P311 (AMI NEC_ P311 (AMI NEC_ ACID

[0017] Docket No.: 066461.11126 / 25WO1 NEC_ (AMI NEC_ ACID NEC_ ACID NEC_ P310 (AMI NEC_ P310 (AMI NEC_ P310 (AMI NEC_ ACID

[0018] Docket No.: 066461.11126 / 25WO1 NEC_ ACID NEC_ P310 (AMI NEC_ P311 (AMI NEC_ ACID NEC_ ACID NEC_ ACID DB01 P211 (NUC TGCAAAGAATACGGTATACCTCCAAATGAACAGCCTGAAGCCTGAAGACACGGC

[0019] Docket No.: 066461.11126 / 25WO1 DB01 P210 (NUC DB01 P211 (NUC DB01 P212 (NUC

[0020] Docket No.: 066461.11126 / 25WO1 DB01 P210 (NUC DB01 P212 (NUC NEC_ (NUC NEC_ (NUC GGGGGGTACCTATTACGCGGACTCGGTAAAAGGCCGTTTTACGATCAGTCGTGAT

[0021] Docket No.: 066461.11126 / 25WO1 NEC_ P311 (NUC NEC_ (NUC NEC_ P311 (NUC

[0022] Docket No.: 066461.11126 / 25WO1 NEC_ P311 (NUC NEC_ (NUC NEC_ (NUC NEC_ (NUC GTTCTACATATTACGCGGACTCGGTAAAAGGCCGTTTTACGATCAGTCGTGATAA

[0023] Docket No.: 066461.11126 / 25WO1 NEC_ (NUC NEC_ P310 (NUC NEC_ P310 (NUC

[0024] Docket No.: 066461.11126 / 25WO1 NEC_ P310 (NUC NEC_ (NUC NEC_ (NUC NEC_ P310 (NUC AGGCTCTACCCGTTACGCGGACTCGGTAAAAGGCCGTTTTACGATCAGTCGTGAT

[0025] Docket No.: 066461.11126 / 25WO1 NEC_ P311 (NUC NEC_ (NUC NEC_ (NUC

[0026] Docket No.: 066461.11126 / 25WO1 NEC_ (NUC SPAC CD8 (AA 1 CD28 (AA 1 TRA CXC NUC SEQU CXC NUC SEQU

[0027] Docket No.: 066461.11126 / 25WO1 CXC NUC SEQU CXC NUC SEQU CXC NUC SEQU CXC NUC SEQU CXC NUC SEQU LINK Whitl (G4S) Linke Linke

[0028] Docket No.: 066461.11126 / 25WO1 Linke Linke Linke Linke Linke Linke Linke Linke Linke Linke Linke Linke Linke Linke Linke Linke

[0029] Docket No.: 066461.11126 / 25WO1 Linke Linke Linke Linke Linke Linke Linke Linke Linke Linke Linke Linke Linke Linke Linke

[0030] Docket No.: 066461.11126 / 25WO1 Whitl (nucle Whitl (nucle GPCR ADD AMD (AMI CXC AND END FUSE TER AMD (NUC CXC AND END FUSE TER TGTTCAATATCAACTTCTACGCTGGAGCTCTGCTGCTGGCCTGCATCAGCTTCGAT

[0031] Docket No.: 066461.11126 / 25WO1 AMD (AMI CXC AND C 3 FLSAHHDERLNATHCQYNFPQVGRTALRVLQLVAGFLLPLLVMAYCYAHILAVLLV

[0032] Docket No.: 066461.11126 / 25WO1 END FUSE TER AND END INSE INTR LOO AMD (NUC CXC AND END FUSE TER AND END INSE INTR LOO TGGTGGATATCCTGATGGACCTGGGCGCCCTCGCACGGAACTGTGGCAGGGAATC

[0033] Docket No.: 066461.11126 / 25WO1 AMD (AMI CXC AND END FUSE TER AND END INSE INTR LOO

[0034] Docket No.: 066461.11126 / 25WO1 AMD (NUC CXC AND END FUSE TER AND END INSE INTR LOO GGAGTACGATGTTTTGGACAAGCGACGTGGCCGGGACCCTGAGATGGGGGGAAA

[0035] Docket No.: 066461.11126 / 25WO1 AMD (AMI CXC END FUSE TER AND END INSE INTR LOO AMD (NUC CXC END FUSE TER AND END INSER O GGTAGCGGGAGCTCTGTTCAATATCAACTTCTACGCTGGAGCTCTGCTGCTGGCC

[0036] Docket No.: 066461.11126 / 25WO1 INTR LOO AMD (AMI CXC END FUSE TERM , VTSGLGYMHCCLNPLLYAFVGVKFRERMWMLLLRLGCPNQRGLQRQPSSSRRDSS

[0037] Docket No.: 066461.11126 / 25WO1 AND END INSE INTR LOO AMD (NUC CXC END FUSE TER AND END INSE INTR LOO CTCCTCCTTAGGTTGGGCTGCCCCAATCAGAGGGGCTTGCAGCGACAACCTTCCT

[0038] Docket No.: 066461.11126 / 25WO1 AMD (AMI CXC AND END FUSE TER AND END INSE INTR LOO AMD (NUC CXC AND C 3 GCGGCTGTCCTTCTGAGTGCCCTGTACCTGCTCCGGAGGGACCAGAGGCTGCCCC

[0039] Docket No.: 066461.11126 / 25WO1 END FUSE TER AND END INSE INTR LOO

[0040] Docket No.: 066461.11126 / 25WO1 AMD (AMI CXC AND END FUSE TER OX40 INSE ICL1, INSE AMD (NUC CXC AND END FUSE TER OX40 INSE ICL1, INSE CTGAATGCCACCCACTGCCAGTATAATTTCCCTCAAGTAGGTCGGACCGCCCTTC

[0041] Docket No.: 066461.11126 / 25WO1 ANTI 9G8- EGFR (AMI WILD-TYPE CXCR3 TLLVLTLPLWAVDAAVQWVFGSGLCKVAGALFNINFYAGALLLACISFDRYLNIVHA

[0042] Docket No.: 066461.11126 / 25WO1 RECE ANTI FUSE TER 9G8- EGFR (NUC WILD RECE ANTI FUSE TER GGTTGTTGCTTTCGCCTTATGCTGGACCCCATACCATTTAGTTGTGCTGGTGGATA

[0043] Docket No.: 066461.11126 / 25WO1 AMD BIND (AMI 9G8- ANTI FUSE TER AND AND END FUSE TER AMD BIND (NUC 9G8- ANTI FUSE TERM INAL TAIL, TTGGAGAACTTCAGTTCATCCTACGATTACGGAGAGAACGAGTCTGATAGTTGTT

[0044] Docket No.: 066461.11126 / 25WO1 AND AND END FUSE TER AMD BIND (AMI

[0045] Docket No.: 066461.11126 / 25WO1 9G8- ANTI FUSE TER AND AND END FUSE TER AND END INSE INTR LOO AMD BIND (NUC 9G8- ANTI FUSE TER AND AND END FUSE O C- GGCTGTCCTTCTGAGTCGAAGCAAGCGGAGCCGGCTGCTGCACTCCGACTACATG

[0046] Docket No.: 066461.11126 / 25WO1 TER AND END INSE INTR LOO

[0047] Docket No.: 066461.11126 / 25WO1 AMD BIND (AMI 9G8- ANTI FUSE TER AND AND END FUSE TER AND END INSE INTR LOO AMD BIND (NUC 9G8- ANTI FUSE TER AND WITH 4-1BB

[0048] Docket No.: 066461.11126 / 25WO1 AND END FUSE TER AND END INSE INTR LOO GAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGAT

[0049] Docket No.: 066461.11126 / 25WO1 AMD BIND (AMI 9G8- ANTI FUSE TER AND END FUSE TER AND END INSE INTR LOO AMD BIND (NUC 9G8- ANTI - G 9G8 GCTGTGTACTATTGCGCTGCAGGGTATCAGATAAATAGCGGTAACTATAATTTCA

[0050] Docket No.: 066461.11126 / 25WO1 FUSE TER AND END FUSE TER AND END INSE INTR LOO CTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACA

[0051] Docket No.: 066461.11126 / 25WO1 AMD BIND (AMI 9G8- ANTI FUSE TER AND END FUSE TER AND END INSE INTR LOO AMD BIND (NUC 9G8- ANTI - G 9G8 GCTGTGTACTATTGCGCTGCAGGGTATCAGATAAATAGCGGTAACTATAATTTCA

[0052] Docket No.: 066461.11126 / 25WO1 FUSE TER AND END FUSE TER AND END INSE INTR LOO CAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGT

[0053] Docket No.: 066461.11126 / 25WO1 AMD BIND (AMI 9G8- ANTI FUSE TER AND AND END FUSE TER AND END INSE INTR LOO AMD BIND (NUC AATTCAAAAAATACCTTATACCTGCAAATGAACAGTCTGCGGGCCGAAGATACG

[0054] Docket No.: 066461.11126 / 25WO1 9G8- ANTI FUSE TER AND AND END FUSE TER AND END INSE INTR LOO CAGAAGAAGAAGAAGGAGGATGTGAACTGAGAGTGAAGTTCAGCAGGAGCGCA

[0055] Docket No.: 066461.11126 / 25WO1 AMD BIND (AMI 9G8- ANTI FUSE TER AND AND END FUSE TER OX40 INSE ICL1, INSE

[0056] Docket No.: 066461.11126 / 25WO1 AMD BIND (NUC 9G8- ANTI FUSE TER AND AND END FUSE TER OX40 INSE ICL1, INSE ACCATTTAGTTGTGCTGGTGGATATCCTGATGGACCTGGGCGCCCTCGCACGGAA

[0057] Docket No.: 066461.11126 / 25WO1

[0058] Docket No.: 066461.11126 / 25WO1 SEQ ID NOs: 234-247 of the present disclosure provides GPCR scaffolds sequences (both amino acid and nucleotide sequences) to which a sequence encoding one or more antigen binding domains may be added. For example, SEQ ID NO: 249 provides a GPCR CAR wherein the anti-EGFR antigen binding domain according to SEQ ID NO: 158 is added to the GPCR scaffold of SEQ ID NO: 234 of the present disclosure. In another example, SEQ ID NO: 250 provides a GPCR CAR wherein the anti-EGFR antigen binding domain according to SEQ ID NO: 158 is added to the GPCR scaffold of SEQ ID NO: 235 of the present disclosure. Generally, the antigen binding domain may be disposed on an N-terminal end of a GPCR scaffold or along an extracellular loop of the GPCR scaffold. In contrast, an intracellular signaling domain or co-stimulatory domain may be disposed on an C-terminal end of a GPCR scaffold or along an intracellular loop of the GPCR scaffold. A person of skill in the art will appreciate that any antigen binding domain, intracellular signaling domain, costimulatory domain of the present disclosure or combination thereof may be added to a GOCR scaffold of the present disclosure. In certain embodiments, a GPCR CAR of the present disclosure comprises (i) a GPCR scaffold comprising amino acids having at least about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% sequence identity to one or more of SEQ ID NOs: 234-240, and (ii) an extracellular domain comprising an antigen binding domain comprising amino acids having at least about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% sequence identity to one or more of SEQ ID NOs: 2, 4, and 158-193. In other embodiments, a GPCR CAR of the present disclosure comprises (i) a GPCR scaffold comprising (a) amino acids having at least about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% sequence identity to one or more of SEQ ID NOs: 234-240, or (b) nucleotides having at least about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% sequence identity to one or more of SEQ ID NOs: 241-247, and (ii) an extracellular domain comprising an antigen binding domain selected from Table 1. In certain embodiments, the GPCR CAR comprises an anti-EGFR antigen binding domain, and the CAR comprises amino acids having at least about 90%, about 91%, about 92%, about 93%, Docket No.: 066461.11126 / 25WO1 about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% sequence identity to one of SEQ ID NOs: 248-255. In other embodiments, the GPCR CAR comprises an anti-EGFR antigen binding domain, and the CAR comprises is encoded by a polynucleotide comprising nucleotides having at least about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% sequence identity to one of SEQ ID NOs: 256-263. III. Artificial Cell Death Polypeptide According to embodiments of the application, an iPSC or a derivative cell thereof may comprise a second exogenous polynucleotide encoding an artificial cell death polypeptide. As used herein, the term "artificial cell death polypeptide” refers to an engineered protein designed to prevent potential toxicity or otherwise adverse effects of a cell therapy. The artificial cell death polypeptide could mediate induction of apoptosis, inhibition of protein synthesis, DNA replication, growth arrest, transcriptional and post- transcriptional genetic regulation and / or antibody-mediated depletion. In some instance, the artificial cell death polypeptide is activated by an exogenous molecule, e.g. an antibody, that when activated, triggers apoptosis and / or cell death of a therapeutic cell. In certain embodiments, an artificial cell death polypeptide comprises an inactivated cell surface receptor that comprises an epitope specifically recognized by an antibody, particularly a monoclonal antibody, which is also referred to herein as a monoclonal antibody-specific epitope. When expressed by iPSCs or derivative cells thereof, the inactivated cell surface receptor is signaling inactive or significantly impaired, but can still be specifically recognized by an antibody. The specific binding of the antibody to the inactivated cell surface receptor enables the elimination of the iPSCs or derivative cells thereof by ADCC and / or ADCP mechanisms, as well as, direct killing with antibody drug conjugates with toxins or radionuclides. In certain embodiments, the inactivated cell surface receptor comprises an epitope that is selected from epitopes specifically recognized by an antibody, including but not limited to, ibritumomab, tiuxetan, muromonab-CD3, tositumomab, abciximab, basiliximab, brentuximab vedotin, cetuximab, infliximab, rituximab, alemtuzumab, Docket No.: 066461.11126 / 25WO1 bevacizumab, certolizumab pegol, daclizumab, eculizumab, efalizumab, gemtuzumab, natalizumab, omalizumab, palivizumab, polatuzumab vedotin, ranibizumab, tocilizumab, trastuzumab, vedolizumab, adalimumab, belimumab, canakinumab, denosumab, golimumab, ipilimumab, avelumab, ofatumumab, panitumumab, or ustekinumab. In certain embodiments, the inactivated cell surface receptor comprises an epitope that is specifically recognized by cetuximab. In certain embodiments, the inactivated cell surface receptor comprises an epitope that is specifically recognized by trastuzumab. In certain embodiments, the inactivated cell surface receptor comprises an epitope that is specifically recognized by bevacizumab. In certain embodiments, the inactivated cell surface receptor comprises an epitope that is specifically recognized by avelumab. In certain embodiments, the inactivated cell surface receptor comprises an epitope that is specifically recognized by ipilimumab. Epidermal growth factor receptor, also known as EGFR, ErbB1 and HER1, is a cell-surface receptor for members of the epidermal growth factor family of extracellular ligands. As used herein, “truncated EGFR,” “tEGFR,” “short EGFR” or “sEGFR” refers to an inactive EGFR variant that lacks the EGF-binding domains and the intracellular signaling domains of the EGFR. An exemplary tEGFR variant contains residues 322-333 of domain 2, all of domains 3 and 4 and the transmembrane domain of the native EGFR sequence containing the cetuximab binding epitope. Expression of the tEGFR variant on the cell surface enables cell elimination by an antibody that specifically binds to the tEGFR, such as cetuximab (Erbitux®), as needed. Due to the absence of the EGF-binding domains and intracellular signaling domains, tEGFR is inactive when expressed by iPSCs or derivative cell thereof. An exemplary inactivated cell surface receptor of the application comprises a tEGFR variant. In certain embodiments, expression of the inactivated cell surface receptor in an engineered immune cell expressing a chimeric antigen receptor (CAR) induces cell suicide of the engineered immune cell when the cell is contacted with an anti-EGFR antibody. Methods of using inactivated cell surface receptors are described in WO2019 / 070856, WO2019 / 023396, WO2018 / 058002, the disclosure of which is incorporated herein by reference. For example, a subject who has previously received an engineered immune cell of the present disclosure that comprises a heterologous Docket No.: 066461.11126 / 25WO1 polynucleotide encoding an inactivated cell surface receptor comprising a tEGFR variant can be administered an anti-EGFR antibody in an amount effective to ablate in the subject the previously administered engineered immune cell. In certain embodiments, the anti-EGFR antibody is cetuximab, matuzumab, necitumumab or panitumumab, preferably the anti-EGFR antibody is cetuximab. In certain embodiments, the tEGFR variant comprises or consists of an amino acid sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, identical to SEQ ID NO: 71, preferably the amino acid sequence of SEQ ID NO: 71. In some embodiments, the inactivated cell surface receptor comprises one or more epitopes of CD79b, such as an epitope specifically recognized by polatuzumab vedotin. In certain embodiments, the CD79b epitope comprises or consists of an amino acid sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, identical to SEQ ID NO: 78, preferably the amino acid sequence of SEQ ID NO: 78. In some embodiments, the inactivated cell surface receptor comprises one or more epitopes of CD20, such as an epitope specifically recognized by rituximab. In certain embodiments, the CD20 epitope comprises or consists of an amino acid sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, identical to SEQ ID NO: 80, preferably the amino acid sequence of SEQ ID NO: 80. In some embodiments, the inactivated cell surface receptor comprises one or more epitopes of Her 2 receptor or ErbB, such as an epitope specifically recognized by trastuzumab. In certain embodiments, the monoclonal antibody-specific epitope comprises or consists of an amino acid sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, identical to SEQ ID NO: 82, preferably the amino acid sequence of SEQ ID NO: 82. IV. Cytokine Expression In some embodiments the iPSC cell or a derivative cell thereof optionally comprises an exogenous polynucleotide encoding a cytokine, such as interleukin-15 or interleukin-2. Docket No.: 066461.11126 / 25WO1 As used herein “Interleukin-15” or “IL-15” refers to a cytokine that regulates T and NK cell activation and proliferation, or a functional portion thereof. A “functional portion” (“biologically active portion”) of a cytokine refers to a portion of the cytokine that retains one or more functions of full length or mature cytokine. Such functions for IL-15 include the promotion of NK cell survival, regulation of NK cell and T cell activation and proliferation as well as the support of NK cell development from hematopoietic stem cells. As will be appreciated by those of skill in the art, the sequence of a variety of IL-15 molecules are known in the art. In certain embodiments, the IL-15 is a wild-type IL-15. In certain embodiments, the IL-15 is a human IL-15. In certain embodiments, the IL-15 comprises an amino acid sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, identical to SEQ ID NO: 72, preferably the amino acid sequence of SEQ ID NO: 72. In some embodiments, the IL-15 is a membrane bound form, where all or a functional portion of the IL-15 protein is fused to all or a portion of a transmembrane protein that anchors the expressed IL-15 as a cell membrane-bound polypeptide (mbIL15)”, for example the construct described in US Patent US9629877B2, hereby incorporated by reference into the present application. As used herein “Interleukin-2” refers to a cytokine that regulates T and NK cell activation and proliferation, or a functional portion thereof. In certain embodiments, the IL-2 is a wild-type IL-2. In certain embodiments, the IL-2 is a human IL-2. In certain embodiments, the IL-2 comprises an amino acid sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, identical to SEQ ID NO: 76, preferably the amino acid sequence of SEQ ID NO: 76. In certain embodiments, an inactivated cell surface receptor comprises a monoclonal antibody-specific epitope operably linked to a cytokine, preferably by an autoprotease peptide sequence. Examples of the autoprotease peptide include, but are not limited to, a peptide sequence selected from the group consisting of porcine teschovirus-1 2A (P2A), a foot-and-mouth disease virus (FMDV) 2A (F2A), an Equine Rhinitis A Virus (ERAV) 2A (E2A), a Thosea asigna virus 2A (T2A), a cytoplasmic polyhedrosis virus 2A (BmCPV2A), a Flacherie Virus 2A (BmIFV2A), and a combination thereof. In Docket No.: 066461.11126 / 25WO1 one embodiment, the autoprotease peptide is an autoprotease peptide of porcine tesehovirus-12A (P2A). In certain embodiments, the autoprotease peptide comprises an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, identical to SEQ ID NO: 73, preferably the amino acid sequence of SEQ ID NO: 73. In certain embodiments, an inactivated cell surface receptor comprises a truncated epithelial growth factor receptor (tEGFR) variant operably linked to an interleukin-15 (IL-15) or IL-2 by an autoprotease peptide sequence. In a particular embodiment, the inactivated cell surface receptor comprises an amino acid sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, identical to SEQ ID NO: 74, preferably the amino acid sequence of SEQ ID NO: 74. In some embodiments, an inactivated cell surface receptor further comprises a signal sequence. In certain embodiments, the signal sequence comprises an amino acid sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, identical to SEQ ID NO: 77, preferably the amino acid sequence of SEQ ID NO: 77. In some embodiments, an inactivated cell surface receptor further comprises a hinge domain. In some embodiments, the hinge domain is derived from CD8. In one embodiment, the CD8 hinge domain comprises the amino acid sequence set forth in SEQ ID NO: 21, or a variant thereof having at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, at least 98 or at least 99%, sequence identity with SEQ ID NO: 21. In certain embodiments, an inactivated cell surface receptor further comprises a transmembrane domain. In some embodiments, the transmembrane domain is derived from CD8. In one embodiment, the CD8 transmembrane domain comprises the amino acid sequence set forth in SEQ ID NO: 23, or a variant thereof having at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, at least 98 or at least 99%, sequence identity with SEQ ID NO: 23. In certain embodiment, an inactivated cell surface receptor comprises one or more epitopes specifically recognized by an antibody in its extracellular domain, a Docket No.: 066461.11126 / 25WO1 transmembrane region and a cytoplasmic domain. In some embodiments, the inactivated cell surface receptor further comprises a hinge region between the epitope(s) and the transmembrane region. In some embodiments, the inactivated cell surface receptor comprises more than one epitopes specifically recognized by an antibody, the epitopes can have the same or different amino acid sequences, and the epitopes can be linked together via a peptide linker, such as a flexible peptide linker have the sequence of (GGGGS)n, wherein n is an integer of 1-8 (SEQ ID NO: 25). In some embodiments, the inactivated cell surface receptor further comprises a cytokine, such as an IL-15 or IL-2. In certain embodiments, the cytokine is in the cytoplasmic domain of the inactivated cell surface receptor. Preferably, the cytokine is operably linked to the epitope(s) specifically recognized by an antibody, directly or indirectly, via an autoprotease peptide sequence, such as those described herein. In some embodiments, the cytokine is indirectly linked to the epitope(s) by connecting to the transmembrane region via the autoprotease peptide sequence. Non-limiting exemplary inactivated cell surface receptor regions and sequences are provided in Table 5.

[0059] Docket No.: 066461.11126 / 25WO1 Table 5. RE tEGF tEGF P2A IL-15 CD79 Signa CD79 CD8 182)

[0060] Docket No.: 066461.11126 / 25WO1 CD8 203) P2A IL-15 CD20 Signa CD20 Linke CD8 182) CD8 203) P2A IL-15 ErbB

[0061] Docket No.: 066461.11126 / 25WO1 Signa ErbB P2A IL-15

[0062] Docket No.: 066461.11126 / 25WO1 In a particular embodiment, the inactivated cell surface receptor comprises an amino acid sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, identical to SEQ ID NO: 79, preferably the amino acid sequence of SEQ ID NO: 79. In a particular embodiment, the inactivated cell surface receptor comprises an amino acid sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, identical to SEQ ID NO: 81, preferably the amino acid sequence of SEQ ID NO: 81. In a particular embodiment, the inactivated cell surface receptor comprises an amino acid sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, identical to SEQ ID NO: 83, preferably the amino acid sequence of SEQ ID NO: 83. V. HLA Expression In one aspect, MHC I and / or MHC II knock-out and / or knock down can be incorporated in the cells for use in “allogeneic” cell therapies, in which cells are harvested from a subject, modified to knock-out or knock-down, e.g., disrupt, B2M, TAP 1, TAP 2, Tapasin, RFXANK, CIITA, RFX5 and RFXAP gene expression, and then returned to a different subject. Knocking out or knocking down the B2M, TAP 1, TAP 2, Tapasin, RFXANK, CIITA, RFX5 and RFXAP genes as described herein can: (1) prevent Graft versus Host response; (2) prevent Host versus Graft response; and / or (3) improve cell safety and efficacy. Accordingly, certain embodiments of the present disclosure comprise independently knocking out and / or knocking down one or more genes selected from the group consisting of B2M, TAP 1, TAP 2, Tapasin, RFXANK, CIITA, RFX5 and RFXAP genes in an iPSC cell. In certain embodiments, a presently disclosed method comprises independently knocking out and / or knocking down two genes selected from the group consisting B2M, TAP 1, TAP 2, Tapasin, RFXANK, CIITA, RFX5 and RFXAP genes in an iPSC cell, in particular, B2M and CIITA to achieve class I and II HLA disruption.. In certain embodiments, an iPSC or derivative cell thereof of the application can be further modified by introducing an exogenous polynucleotide encoding one or more proteins related to immune evasion, such as non- Docket No.: 066461.11126 / 25WO1 classical HLA class I proteins (e.g., HLA-E and HLA-G). In particular, disruption of the B2M gene eliminates surface expression of all MHC class I molecules, leaving cells vulnerable to lysis by NK cells through the “missing self” response. Exogenous HLA-E expression can lead to resistance to NK-mediated lysis (Gornalusse et al., Nat Biotechnol. 2017; 35(8): 765–772). Incorporating MHC I and / or MHC II knock-out and / or knock down in the cells for use in “allogeneic” cell therapies will allow the cell product candidates to escape recognition and destruction by the host immune system. The reduction in allogeneic reactivity enabled by use of this technology will allow repeat dosing of the CAR- modified cell therapies to improve their therapeutic potential. In combination with the extended killing capability of optimized immune cells derived from single genetically engineered cell cloning, the cells will have the capacity for repeat dosing to maximize durability of response and efficacy. Additionally, this technology may permit dosing in patients with limited or no immune preconditioning regimens. Accordingly, in certain embodiments, an iPSC or derivative cell thereof of the application can be further modified by introducing a third exogenous polynucleotide encoding one or more proteins related to immune evasion, such as non-classical HLA class I proteins (e.g., HLA-E and HLA-G). In certain embodiments, the iPSC or derivative cell thereof comprises a third exogenous polypeptide encoding at least one of a human leukocyte antigen E (HLA-E) and human leukocyte antigen G (HLA-G). In a particular embodiment, the HLA-E comprises an amino acid sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, identical to SEQ ID NO: 65, preferably the amino acid sequence of SEQ ID NO: 65. In a particular embodiment, the HLA-G comprises an amino acid sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, identical to SEQ ID NO: 68, preferably SEQ ID NO: 68. In certain embodiments, the third exogenous polynucleotide encodes a polypeptide comprising a signal peptide operably linked to a mature B2M protein that is fused to an HLA-E via a linker. In a particular embodiment, the third exogenous Docket No.: 066461.11126 / 25WO1 polypeptide comprises an amino acid sequence at least sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, identical to SEQ ID NO: 66. In other embodiments, the third exogenous polynucleotide encodes a polypeptide comprising a signal peptide operably linked to a mature B2M protein that is fused to an HLA-G via a linker. In a particular embodiment, the third exogenous polypeptide comprises an amino acid sequence at least sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, identical to SEQ ID NO: 69. VI. Other Optional Genome Edits In one embodiment of the above described cell, the genomic editing at one or more selected sites may comprise insertions of one or more exogenous polynucleotides encoding other additional artificial cell death polypeptides, targeting modalities, receptors, signaling molecules, transcription factors, pharmaceutically active proteins and peptides, drug target candidates, or proteins promoting engraftment, trafficking, homing, viability, self-renewal, persistence, and / or survival of the genome-engineered iPSCs or derivative cells thereof. In some embodiments, the exogenous polynucleotides for insertion are operatively linked to (1) one or more exogenous promoters comprising CMV, EFla, PGK, CAG, UBC, or other constitutive, inducible, temporal-, tissue-, or cell type-specific promoters; or (2) one or more endogenous promoters comprised in the selected sites comprising AAVS1, CLYBL, CCR5, ROSA26, collagen, HTRP, Hll, beta-2 microglobulin, GAPDH, TCR or RUNX1, or other locus meeting the criteria of a genome safe harbor. In some embodiments, the genome-engineered iPSCs generated using the above method comprise one or more different exogenous polynucleotides encoding proteins comprising caspase, thymidine kinase, cytosine deaminase, B-cell CD20, ErbB2 or CD79b wherein when the genome-engineered iPSCs comprise two or more suicide genes, the suicide genes are integrated in different safe harbor locus comprising AAVSl, CCR5, ROSA26, collagen, HTRP, Hll, beta-2 microglobulin, GAPDH, TCR or RUNX1. Other exogenous polynucleotides encoding proteins may include those encoding PET Docket No.: 066461.11126 / 25WO1 reporters, homeostatic cytokines, and inhibitory checkpoint inhibitory proteins such as PD1, PD-L1, CTLA4, and proteins that target the CD47 / signal regulatory protein alpha (SIRPα) axis. In one aspect, the cell may comprise an exogenous polynucleotide encoding a CD16 protein and / or an NKG2D protein, wherein the CD16 protein and the NKG2D protein may be operably linked by an autoprotease peptide as disclosed in co-pending patent application PCT / US23 / 68079. Accordingly, in some aspects, the cells of the present disclosure can comprise genetically engineered iPSCs and cells derived therefrom that exogenously express recombinant CD16 and recombinant NKG2D. The surface receptor CD16 (FcγRIIIA) affects human natural killer (NK) cells during maturation. NK cells bind the Fc portion of IgG via CD16, and execute antibody-dependent cellular cytotoxicity, which is critical for the effectiveness of several anti-tumor monoclonal antibody therapies. NKG2D is a stimulatory / activating receptor that is mostly expressed on cells of the cytotoxic arm of the immune system including NK cells and subsets of T cells. NKG2D is crucial in diverse aspects of innate and adaptive immune functions. In some embodiments, CD16 and NKG2D are expressed from in a single polynucleotide construct as it is advantageous to reduce the number of gene edits of a cell. . In some embodiments, the polynucleotide construct encoding the CD16 protein and the NKG2D protein also includes a polynucleotide sequence encoding an autoprotease peptide or self-cleaving peptide. In some embodiments, an exogenous polynucleotide construct encoding the CD16 protein, the NKG2D protein and the self- cleaving peptide is introduced into the iPSC or derivative cell thereof. The exogenous or isolated polynucleotide construct can be introduced into a gene locus of the iPSC or derivative cell thereof. In some embodiments, the exogenous polynucleotide construct comprises the nucleic acid sequence of SEQ ID NO: 84. In some embodiments, the exogenous polynucleotide construct encodes for the amino acid sequence of SEQ ID NO: 85.In some embodiments, the CD16 protein (which is also referred to as “low affinity immunoglobulin gamma Fc region receptor III-A” or “Fc gamma receptor IIIa”) is a wildtype CD16 protein. In some embodiments, the human wildtype CD16 protein has the amino acid sequence set forth in NCBI Ref. Seq. No. NP_000560.7 or UniProt No. Docket No.: 066461.11126 / 25WO1 P08637. In some instance, the coding sequence of human wildtype CD16 is set forth in NCBI Ref. No. NM_000569.8. In some embodiments, the CD16 protein is a CD16 variant protein. In some instances, the CD16 variant protein has an amino acid sequence having at least 90%, e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to wildtype CD16 such as that of SEQ ID NO: 86. In some instances, the CD16 variant is a high affinity CD16 variant. In other instances, the CD16 variant is a non-cleavable CD16 variant. In some instances, the CD16 variant is a high affinity and non-cleavable CD16 variant. In some embodiments, the CD16 variant comprises one or more amino acid substitutions selected from the group consisting of F158V, F176V, S197P, D205A, S219A, T220A, and any combination thereof. In some embodiments, the CD16 variant has an F158V substitution and one or more substitutions selected from F176V, S197P, D205A, S219A, T220A, and any combination thereof. In one embodiment, the CD16 variant has an F176V substitution and one or more substitutions selected from F158V, S197P, D205A, S219A, T220A, and any combination thereof. In many embodiments, the CD16 variant has an S197P, substitution and one or more substitutions selected from F158V, F176V, D205A, S219A, T220A, and any combination thereof. In various embodiments, the CD16 variant has a D205A substitution and one or more substitutions selected from F158V, F176V, S197P, S219A, T220A, and any combination thereof. In some embodiments, the CD16 variant has a substitution and one or more substitutions selected from F158V, F176V, S197P, D205A, S219A, T220A, and any combination thereof. In some embodiments, the CD16 variant has an S219A substitution and one or more substitutions selected from F158V, F176V, S197P, D205A, T220A, and any combination thereof. In some embodiments, the CD16 variant has a T220A substitution and one or more substitutions selected from F158V, F176V, S197P, D205A, S219A, T220A, and any combination thereof. In some embodiments, the variant CD16 protein has the sequence of SEQ ID NO: 87. In some embodiments, the nucleic acid sequence encoding the variant CD16 protein has the sequence of SEQ ID NO: 88. In some embodiments, the wildtype CD16 protein has the sequence of SEQ ID NO: 86. Docket No.: 066461.11126 / 25WO1 In some embodiments, the NKG2D protein (which is also referred to as NKG2-D type II integral membrane protein, CD314, killer cell lectin-like receptor subfamily K1 member 1 or KLRK1) is a wildtype NKG2D protein. In some embodiments, the human wildtype NKG2D protein has the amino acid sequence set forth in NCBI Ref. Seq. Nos. NP_001186734.1 or NP_031386.2 or UniProt No. P26718. In some instance, the coding sequence of human wildtype NKG2D is set forth in NCBI Ref. Nos. NM_001199805.1 or NM_007360.3. In some embodiments, the NKG2D protein is a NKG2D variant protein. In some instances, the NKG2D variant protein has an amino acid sequence having at least 90%, e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to wildtype NKG2D such as that of SEQ ID NO: 89. In some embodiments, the NKG2D protein has the amino acid sequence of SEQ ID NO: 89. In some embodiments, the nucleic acid sequence encoding the NKG2D protein has sequence of SEQ ID NO: 90. As discussed above, provided herein are constructs containing autoprotease peptide sequences including 2A peptides that can induce ribosomal skipping during translation of an polypeptide.2A peptides function to “cleave” an mRNA transcript by making the ribosome skip the synthesis of a peptide bond at the C-terminus, between the glycine (G) and proline (P) residues, thereby leading to separation between the end of the 2A sequence and the next peptide downstream.2A peptides include, but are not limited to, a porcine tesehovirus-12A (P2A) peptide, a foot-and-mouth disease virus (FMDV) 2A (F2A) peptide, an Equine Rhinitis A Virus (ERAV) 2A (E2A) peptide, a Thosea asigna virus 2A (T2A) peptide, a cytoplasmic polyhedrosis virus 2A (BmCPV2A) peptide, and a Flacherie Virus 2A (BmIFV2A) peptide. An exemplary P2A peptide can include an amino acid sequence having at least 90%, such as 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 91. In some embodiment, the P2A peptide has the amino acid sequence of SEQ ID NO: 91. Another optional genome edit is the insertion of a polynucleotide encoding a a 12 (IL-12) comprising a first polypeptide comprising an IL- 12 alpha subunit p35, a second polypeptide comprising an IL-12 beta subunit p40 and a transmembrane fused to the terminus of the first and / or second IL-12 subunit polypeptide Docket No.: 066461.11126 / 25WO1 as disclosed in co-pending patent application PCT / US23 / 68105. In certain embodiments, the polynucleotide encoding the membrane bound IL-12 is fused to a polynucleotide encoding an ADAM17 protease cleavage site peptide for the activation induced release of the IL-12 through the protease ADAM17. ADAM17 is expressed by activated lymphocytes and is directly involved in the liberation of other immune mediators like TNFa that are similarly presented as a membrane anchored form. When this membrane tethered IL-12 is expressed on engineered iNK or T cells, it remains cell associated. Upon cell activation and the increased expression of ADAM17, the protease cleaves the membrane stalk and releases IL-12 into the extracellular space. This type of regulation ensures that the activities of the IL-12 are confined to spaces surrounding the tumor where the engineered immune cells engage their targets on the tumor cells that cause their activation. Accordingly, the cell of the present disclosure can further comprise (i) an exogenous polynucleotide encoding a membrane-bound interleukin 12 (IL-12) comprising a first polypeptide comprising an IL-12 alpha subunit p35 or a polypeptide at least 90% similar thereto, a second polypeptide comprising an IL-12 beta subunit p40 or a polypeptide at least 90% similar thereto, and a transmembrane domain fused to the terminus of the first and / or second IL-12 subunit polypeptide. In some other embodiments, the genome-engineered iPSCs generated using the method provided herein comprise in / del at one or more endogenous genes associated with targeting modality, receptors, signaling molecules, transcription factors, drug target candidates, immune response regulation and modulation, or proteins suppressing engraftment, trafficking, homing, viability, self-renewal, persistence, and / or survival of the iPSCs or derivative cells thereof. VII. Targeted Genome Editing at Selected Locus in iPSCs According to embodiments of the application, one or more of the exogenous polynucleotides are integrated at one or more loci on the chromosome of an iPSC. Genome editing, or genomic editing, or genetic editing, as used interchangeably herein, is a type of genetic engineering in which DNA is inserted, deleted, and / or replaced in the genome of a targeted cell. Targeted genome editing (interchangeable with “targeted genomic editing” or “targeted genetic editing”) enables insertion, deletion, Docket No.: 066461.11126 / 25WO1 and / or substitution at pre-selected sites in the genome. When an endogenous sequence is deleted or disrupted at the insertion site during targeted editing, an endogenous gene comprising the affected sequence can be knocked-out or knocked-down due to the sequence deletion or disruption. Therefore, targeted editing can also be used to disrupt endogenous gene expression with precision. Similarly used herein is the term “targeted integration,” referring to a process involving insertion of one or more exogenous sequences at pre-selected sites in the genome, with or without deletion of an endogenous sequence at the insertion site. Targeted editing can be achieved either through a nuclease-independent approach, or through a nuclease-dependent approach. In the nuclease-independent targeted editing approach, homologous recombination is guided by homologous sequences flanking an exogenous polynucleotide to be inserted, through the enzymatic machinery of the host cell. Alternatively, targeted editing could be achieved with higher frequency through specific introduction of double strand breaks (DSBs) by specific rare-cutting endonucleases. Such nuclease-dependent targeted editing utilizes DNA repair mechanisms including non-homologous end joining (NHEJ), which occurs in response to DSBs. Without a donor vector containing exogenous genetic material, the NHEJ often leads to random insertions or deletions (in / dels) of a small number of endogenous nucleotides. In comparison, when a donor vector containing exogenous genetic material flanked by a pair of homology arms is present, the exogenous genetic material can be introduced into the genome during homology directed repair (HDR) by homologous recombination, resulting in a “targeted integration.” Available endonucleases capable of introducing specific and targeted DSBs include, but not limited to, zinc-finger nucleases (ZFN), transcription activator-like effector nucleases (TALEN), RNA-guided CRISPR (Clustered Regular Interspaced Short Palindromic Repeats) systems. Additionally, DICE (dual integrase cassette exchange) system utilizing phiC31 and Bxbl integrases is also a promising tool for targeted integration. ZFNs are targeted nucleases comprising a nuclease fused to a zinc finger DNA binding domain. By a “zinc finger DNA binding domain” or “ZFBD” it is meant a Docket No.: 066461.11126 / 25WO1 polypeptide domain that binds DNA in a sequence-specific manner through one or more zinc fingers. A zinc finger is a domain of about 30 amino acids within the zinc finger binding domain whose structure is stabilized through coordination of a zinc ion. Examples of zinc fingers include, but not limited to, C2H2 zinc fingers, C3H zinc fingers, and C4 zinc fingers. A “designed” zinc finger domain is a domain not occurring in nature whose design / composition results principally from rational criteria, e.g., application of substitution rules and computerized algorithms for processing information in a database storing information of existing ZFP designs and binding data. See, for example, U.S. Pat. Nos.6,140,081; 6,453,242; and 6,534,261; see also WO 98 / 53058; WO 98 / 53059; WO 98 / 53060; WO 02 / 016536 and WO 03 / 016496. A “selected” zinc finger domain is a domain not found in nature whose production results primarily from an empirical process such as phage display, interaction trap or hybrid selection. ZFNs are described in greater detail in U.S. Pat. No.7,888,121 and U.S. Pat. No.7,972,854, the complete disclosures of which are incorporated herein by reference. The most recognized example of a ZFN in the art is a fusion of the Fokl nuclease with a zinc finger DNA binding domain. A TALEN is a targeted nuclease comprising a nuclease fused to a TAL effector DNA binding domain. By “transcription activator-like effector DNA binding domain”, “TAL effector DNA binding domain”, or “TALE DNA binding domain” it is meant the polypeptide domain of TAL effector proteins that is responsible for binding of the TAL effector protein to DNA. TAL effector proteins are secreted by plant pathogens of the genus Xanthomonas during infection. These proteins enter the nucleus of the plant cell, bind effector-specific DNA sequences via their DNA binding domain, and activate gene transcription at these sequences via their transactivation domains. TAL effector DNA binding domain specificity depends on an effector-variable number of imperfect 34 amino acid repeats, which comprise polymorphisms at select repeat positions called repeat variable-diresidues (RVD). TALENs are described in greater detail in U.S. Patent Application No.2011 / 0145940, which is herein incorporated by reference. The most recognized example of a TALEN in the art is a fusion polypeptide of the Fokl nuclease to a TAL effector DNA binding domain. Another example of a targeted nuclease that finds use in the subject methods is a targeted Spoll nuclease, a polypeptide comprising a Spol l polypeptide having nuclease Docket No.: 066461.11126 / 25WO1 activity fused to a DNA binding domain, e.g. a zinc finger DNA binding domain, a TAL effector DNA binding domain, etc. that has specificity for a DNA sequence of interest. See, for example, U.S. Application No.61 / 555,857, the disclosure of which is incorporated herein by reference. Additional examples of targeted nucleases suitable for the present application include, but not limited to Bxbl, phiC3 l, R4, PhiBTl, and Wp / SPBc / TP90l-l, whether used individually or in combination. Other non-limiting examples of targeted nucleases include naturally occurring and recombinant nucleases; CRISPR related nucleases from families including cas, cpf, cse, csy, csn, csd, cst, csh, csa, csm, and cmr; restriction endonucleases; meganucleases; homing endonucleases, and the like. As an example, CRISPR / Cas9 requires two major components: (1) a Cas9 endonuclease and (2) the crRNA-tracrRNA complex. When co- expressed, the two components form a complex that is recruited to a target DNA sequence comprising PAM and a seeding region near PAM. The crRNA and tracrRNA can be combined to form a chimeric guide RNA (gRNA) to guide Cas9 to target selected sequences. These two components can then be delivered to mammalian cells via transfection or transduction. As another example, CRISPR / Cpf1 comprises two major components: (1) a CPf1 endonuclease and (2) a crRNA. When co-expressed, the two components form a ribobnucleoprotein (RNP) complex that is recruited to a target DNA sequence comprising PAM and a seeding region near PAM. The crRNA can be combined to form a chimeric guide RNA (gRNA) to guide Cpf1 to target selected sequences. These two components can then be delivered to mammalian cells via transfection or transduction. MAD7 is an engineered Cas12a variant originating from the bacterium Eubacterium rectale that has a preference for 5′-TTTN-3′ and 5′-CTTN-3′ PAM sites and does not require a tracrRNA. See, for example, PCT Publication No. 2018 / 236548, the disclosure of which is incorporated herein by reference. DICE mediated insertion uses a pair of recombinases, for example, phiC31 and Bxbl, to provide unidirectional integration of an exogenous DNA that is tightly restricted to each enzymes’ own small attB and attP recognition sites. Because these target att sites are not naturally present in mammalian genomes, they must be first introduced into the Docket No.: 066461.11126 / 25WO1 genome, at the desired integration site. See, for example, U.S. Application Publication No.2015 / 0140665, the disclosure of which is incorporated herein by reference. One aspect of the present application provides a construct comprising one or more exogenous polynucleotides for targeted genome integration. In one embodiment, the construct further comprises a pair of homologous arms specific to a desired integration site, and the method of targeted integration comprises introducing the construct to cells to enable site specific homologous recombination by the cell host enzymatic machinery. In another embodiment, the method of targeted integration in a cell comprises introducing a construct comprising one or more exogenous polynucleotides to the cell, and introducing a ZFN expression cassette comprising a DNA-binding domain specific to a desired integration site to the cell to enable a ZFN-mediated insertion. In yet another embodiment, the method of targeted integration in a cell comprises introducing a construct comprising one or more exogenous polynucleotides to the cell, and introducing a TALEN expression cassette comprising a DNA-binding domain specific to a desired integration site to the cell to enable a TALEN-mediated insertion. In another embodiment, the method of targeted integration in a cell comprises introducing a construct comprising one or more exogenous polynucleotides to the cell, introducing a Cpf1 expression cassette, and a gRNA comprising a guide sequence specific to a desired integration site to the cell to enable a Cpf1-mediated insertion. In another embodiment, the method of targeted integration in a cell comprises introducing a construct comprising one or more exogenous polynucleotides to the cell, introducing a Cas9 expression cassette, and a gRNA comprising a guide sequence specific to a desired integration site to the cell to enable a Cas9-mediated insertion. In still another embodiment, the method of targeted integration in a cell comprises introducing a construct comprising one or more att sites of a pair of DICE recombinases to a desired integration site in the cell, introducing a construct comprising one or more exogenous polynucleotides to the cell, and introducing an expression cassette for DICE recombinases, to enable DICE-mediated targeted integration. Sites for targeted integration include, but are not limited to, genomic safe harbors, which are intragenic or extragenic regions of the human genome that, theoretically, are able to accommodate predictable expression of newly integrated DNA without adverse Docket No.: 066461.11126 / 25WO1 effects on the host cell or organism. In certain embodiments, the genome safe harbor for the targeted integration is one or more loci of genes selected from the group consisting of AAVS1, CLYBL, CCR5, ROSA26, collagen, HTRP, Hll, GAPDH, TCR and RUNX1 genes. In other embodiments, the site for targeted integration is selected for deletion or reduced expression of an endogenous gene at the insertion site. As used herein, the term “deletion” with respect to expression of a gene refers to any genetic modification that abolishes the expression of the gene. Examples of “deletion” of expression of a gene include, e.g., a removal or deletion of a DNA sequence of the gene, an insertion of an exogenous polynucleotide sequence at a locus of the gene, and one or more substitutions within the gene, which abolishes the expression of the gene. Genes for target deletion include, but are not limited to, genes of major histocompatibility complex (MHC) class I and MHC class II proteins. Multiple MHC class I and class II proteins must be matched for histocompatibility in allogeneic recipients to avoid allogeneic rejection problems. “MHC deficient”, including MHC-class I deficient, or MHC-class II deficient, or both, refers to cells that either lack, or no longer maintain, or have reduced level of surface expression of a complete MHC complex comprising a MHC class I protein heterodimer and / or a MHC class II heterodimer, such that the diminished or reduced level is less than the level naturally detectable by other cells or by synthetic methods. MHC class I deficiency can be achieved by functional deletion of any region of the MHC class I locus (chromosome 6p2l), or deletion or reducing the expression level of one or more MHC class-I associated genes including, not being limited to, beta-2 microglobulin (B2M) gene, TAP 1 gene, TAP 2 gene and Tapasin genes. For example, the B2M gene encodes a common subunit essential for cell surface expression of all MHC class I heterodimers. B2M null cells are MHC-I deficient. MHC class II deficiency can be achieved by functional deletion or reduction of MHC-II associated genes including, not being limited to, RFXANK, CIITA, RFX5 and RFXAP. CIITA is a transcriptional coactivator, functioning through activation of the transcription factor RFX5 required for class II protein expression. CIITA null cells are MHC-II deficient. In certain embodiments, one or more of the exogenous polynucleotides are integrated at one or more loci of genes selected from the group consisting of B2M, TAP Docket No.: 066461.11126 / 25WO1 1, TAP 2, Tapasin, RFXANK, CIITA, RFX5 and RFXAP genes to thereby delete or reduce the expression of the gene(s) with the integration. Other genes that may be targeted for deletion include NKG2A, CD38, CD70 and CD33. In certain embodiments, the exogenous polynucleotides are integrated at one or more loci on the chromosome of the cell, preferably the one or more loci are of genes selected from the group consisting of AAVS1, CLYBL, CCR5, ROSA26, collagen, HTRP, Hl l, GAPDH, RUNX1, B2M, TAPI, TAP2, Tapasin, NLRC5, CIITA, RFXANK, CIITA, RFX5, RFXAP, TCR a or b constant region, NKG2A, NKG2D, CD33, CD38, CIS, CBL-B, SOCS2, PD1, CTLA4, LAG3, TIM3, or TIGIT genes, provided at least one of the one or more loci is of a MHC gene, such as a gene selected from the group consisting of B2M, TAP 1, TAP 2, Tapasin, RFXANK, CIITA, RFX5 and RFXAP genes. Preferably, the one or more exogenous polynucleotides are integrated at a locus of an MHC class-I associated gene, such as a beta-2 microglobulin (B2M) gene, TAP 1 gene, TAP 2 gene or Tapasin gene; and at a locus of an MHC-II associated gene, such as a RFXANK, CIITA, RFX5, RFXAP, or CIITA gene; and optionally further at a locus of a safe harbor gene selected from the group consisting of AAVS1, CLYBL, CCR5, ROSA26, collagen, HTRP, Hll, GAPDH, TCR and RUNX1 genes. More preferably, the one or more of the exogenous polynucleotides are integrated at the loci of CIITA, AAVS1 and B2M genes. In certain embodiments, (i) the first exogenous polynucleotide is integrated at a locus of AAVS1 gene or CLYBL gene; (ii) the second exogenous polypeptide is integrated at a locus of CIITA gene; and (iii) the third exogenous polypeptide is integrated at a locus of B2M gene; wherein integrations of the exogenous polynucleotides delete or reduce expression of CIITA and B2M genes. In certain embodiments, (i) the second exogenous polynucleotide comprises the polynucleotide sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 75; and (ii) the third exogenous polynucleotide comprises the polynucleotide sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 67. Docket No.: 066461.11126 / 25WO1 In certain embodiments, (i) the second exogenous polynucleotide comprises the polynucleotide sequence of SEQ ID NO: 75; and (ii) the third exogenous polynucleotide comprises the polynucleotide sequence of SEQ ID NO: 67. VIII. Derivative Cells In another aspect, the present disclosure relates to a cell derived from differentiation of an iPSC, a derivative cell. As described above, the genomic edits introduced into the iPSC are retained in the derivative cell. In certain embodiments of the derivative cell obtained from iPSC differentiation, the derivative cell is a hematopoietic cell, including, but not limited to, HSCs (hematopoietic stem and progenitor cells), hematopoietic multipotent progenitor cells, T cell progenitors, NK cell progenitors, T cells, NKT cells, NK cells, B cells, antigen presenting cells (APC), monocytes and macrophages. In certain embodiments, the derivative cell is an immune effector cell, such as a NK cell or a T cell. In certain embodiments, the application provides a natural killer (NK) cell or a T cell comprising: (i) a first exogenous polynucleotide encoding a chimeric antigen receptor (CAR); (ii) a second exogenous polynucleotide encoding a truncated epithelial growth factor receptor (tEGFR) variant and an interleukin 15 (IL-15), wherein the tEGFR variant and IL-15 are operably linked by an autoprotease peptide sequence, such as autoprotease peptide sequence of porcine tesehovirus-12A (P2A); and (iii) a deletion or reduced expression of an MHC class I associated gene and an MHC class II associated gene, such as an MHC class-I associated gene selected from the group consisting of a B2M gene, TAP 1 gene, TAP 2 gene and Tapasin gene, and an MHC-II associated gene selected from the group consisting of a RFXANK gene, CIITA gene, RFX5 gene, RFXAP gene, and CIITA gene, preferably the B2M gene and CIITA gene. In certain embodiments, the NK cell or T cell further comprises a third exogenous polynucleotide encoding at least one of a human leukocyte antigen E (HLA-E) and a human leukocyte antigen G (HLA-G). Also provided is a NK cell or a T cell comprising: (i) a first exogenous polynucleotide encoding a chimeric antigen receptor (CAR); (ii) a second exogenous Docket No.: 066461.11126 / 25WO1 polynucleotide encoding a truncated epithelial growth factor receptor (tEGFR) variant having the amino acid sequence of SEQ ID NO: 71, an autoprotease peptide having the amino acid sequence of SEQ ID NO: 73, and interleukin 15 (IL-15) having the amino acid sequence of SEQ ID NO: 72; and (iii) a third exogenous polynucleotide encoding a human leukocyte antigen E (HLA-E) having the amino acid sequence of SEQ ID NO: 66; wherein the first, second and third exogenous polynucleotides are integrated at loci of AAVS1, CIITA and B2M genes, respectively, to thereby delete or reduce expression of CIITA and B2M. In certain embodiments, the first exogenous polynucleotide encodes a CAR targeting a tumor antigen; the second exogenous polynucleotide comprises the polynucleotide sequence of SEQ ID NO: 75; and the third exogenous polynucleotide comprises the polynucleotide sequence of SEQ ID NO: 67. Also provided is a CD34+ hematopoietic progenitor cell (HPC) derived from an induced pluripotent stem cell (iPSC) comprising: (i) a first exogenous polynucleotide encoding a chimeric antigen receptor (CAR); (ii) a second exogenous polynucleotide encoding an inactivated cell surface receptor that comprises a monoclonal antibody- specific epitope and an interleukin 15 (IL-15), wherein the inactivated cell surface receptor and IL-15 are operably linked by an autoprotease peptide sequence; and (iii) a deletion or reduced expression of one or more of B2M, TAP 1, TAP 2, Tapasin, RFXANK, CIITA, RFX5 and RFXAP genes. In certain embodiments, the CD34+ HPC further comprises a third exogenous polynucleotide encoding a human leukocyte antigen E (HLA-E) and / or human leukocyte antigen G (HLA-G). In certain embodiments, the CAR comprises (i) a signal peptide; (ii) an extracellular domain comprising a binding domain that specifically binds to a tumor antigen; (iii) a hinge region; (iv) a transmembrane domain; (v) an intracellular signaling domain; and (vi) a co-stimulatory domain, such as a co-stimulatory domain comprising a CD28 signaling domain. Also provided is a method of manufacturing the derivative cell. The method comprises differentiating the iPSC under conditions for cell differentiation to thereby obtain the derivative cell. Docket No.: 066461.11126 / 25WO1 An iPSC of the application can be differentiated by any method known in the art. Exemplary methods are described in US8846395, US8945922, US8318491, WO2010 / 099539, WO2012 / 109208, WO2017 / 070333, WO2017 / 179720, WO2016 / 010148, WO2018 / 048828, WO2019 / 157597, WO2022 / 120334, WO2022 / 133169, WO2022 / 216624, WO2022 / 216514, and WO2022 / 216524, each of which are herein incorporated by reference in its entirety. The differentiation protocol may use feeder cells or may be feeder-free. As used herein, “feeder cells” or “feeders” are terms describing cells of one type that are co-cultured with cells of a second type to provide an environment in which the cells of the second type can grow, expand, or differentiate, as the feeder cells provide stimulation, growth factors and nutrients for the support of the second cell type. In another embodiment of the present disclosure, the iPSC derivative cells are NK cells which are prepared by a method of differentiating an iPSC into an NK cell by subjecting the cells to a differentiation protocol including the addition of recombinant human IL-12p70 for the final 24 hours of culture. By including the IL-12 in the differentiation protocol, cells that are primed with IL-12 demonstrate more rapid cell killing compared to those that are differentiated in the absence of IL-12. In addition, the cells differentiated using the IL-12 conditions demonstrate improved cancer cell growth inhibition. IX. Polynucleotides, vectors, and host cells (1) Nucleic acids encoding a GPCR CAR In another general aspect, the present disclosure relates to an isolated nucleic acid encoding a chimeric antigen receptor (CAR) having a GPCR scaffold (GPCR CAR) useful for embodiments of the application. It will be appreciated by those skilled in the art that the coding sequence of a GPCR CAR can be changed (e.g., replaced, deleted, inserted, etc.) without changing the amino acid sequence of the protein. Accordingly, it will be understood by those skilled in the art that nucleic acid sequences encoding GPCR CARs of the application can be altered without changing the amino acid sequences of the proteins. Docket No.: 066461.11126 / 25WO1 In another general aspect, the application provides a vector comprising a polynucleotide sequence encoding a GPCR CAR useful for embodiments of the application. Any vector known to those skilled in the art in view of the present disclosure can be used, such as a plasmid, a cosmid, a phage vector or a viral vector. In some embodiments, the vector is a recombinant expression vector such as a plasmid. The vector can include any element to establish a conventional function of an expression vector, for example, a promoter, ribosome binding element, terminator, enhancer, selection marker, and origin of replication. The promoter can be a constitutive, inducible, or repressible promoter. A number of expression vectors capable of delivering nucleic acids to a cell are known in the art and can be used herein for production of a GPCR CAR in the cell. Conventional cloning techniques or artificial gene synthesis can be used to generate a recombinant expression vector according to embodiments of the application. In a particular aspect, the application provides vectors for targeted integration of a GPCR CAR useful for embodiments of the application. In certain embodiments, the vector comprises an exogenous polynucleotide having, in the 5’ to 3’ order, (a) a promoter; (b) a polynucleotide sequence encoding a GPCR CAR according to an embodiment of the application; and (c) a terminator / polyadenylation signal. In certain embodiments, the promoter is a CAG promoter. In certain embodiments, the CAG promoter comprises the polynucleotide sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100%, identical to SEQ ID NO: 63. Other promoters can also be used, examples of which include, but are not limited to, EF1a, UBC, CMV, SV40, PGK1, and human beta actin. In certain embodiments, the terminator / polyadenylation signal is a SV40 signal. In certain embodiments, the SV40 signal comprises the polynucleotide sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100%, identical to SEQ ID NO: 64. Other terminator sequences can also be used, examples of which include, but are not limited to, BGH, hGH, and PGK. In some embodiments, the vector further comprises a left homology arm and a right homology arm flanking the exogenous polynucleotide. As used herein, “left homology arm” and “right homology arm” refers to a pair of nucleic acid sequences that flank an exogenous polynucleotide and facilitate the integration of the exogenous Docket No.: 066461.11126 / 25WO1 polynucleotide into a specified chromosomal locus. Sequences of the left and right arm homology arms can be designed based on the integration site of interest. In some embodiments, the left or right arm homology arm is homologous to the left or right side sequence of the integration site. In certain embodiments, the left homology arm comprises the polynucleotide sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100%, identical to one of SEQ ID NOs: 92-103, 130, and 132. In certain embodiments, the right homology arm comprises the polynucleotide sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100%, identical to one of SEQ ID NOs: 104-115, 131, and 133. Table 6 provides an exemplary list of homology arm sequences and corresponding guide sequences for facilitating integration of an exogenous polynucleotide at various loci.

[0063] Docket No.: 066461.11126 / 25WO1 Table 6. Loc AAV (PPP 12C CCCGTGTCTGGGTCCTCTCCGGGCA TTCCTCCGCCCAGAGCAGGGTCCCGC

[0064] Docket No.: 066461.11126 / 25WO1

[0065] Docket No.: 066461.11126 / 25WO1 B2 TCTCCAGTGACAGAAGATACTGCTA TTTCTAACCATTTTAGACATTTGTTAG

[0066] Docket No.: 066461.11126 / 25WO1

[0067] Docket No.: 066461.11126 / 25WO1 CIIT ex1 GGGGCCACCTTGCAGGGAGAGTTT TTTATTGGCATTGCTGTTGTTGGATGG

[0068] Docket No.: 066461.11126 / 25WO1

[0069] Docket No.: 066461.11126 / 25WO1 CIIT ex5 AACTGGACCAGTATGTCTTCCAGGA TGAGATTTTAAAAGTGCAAAGAATAT

[0070] Docket No.: 066461.11126 / 25WO1

[0071] Docket No.: 066461.11126 / 25WO1 CD7 GCCGGACATCCCCAGAGAGGGGCA GGTCTTTTCTTCCAGTGGGACGTAGC

[0072] Docket No.: 066461.11126 / 25WO1

[0073] Docket No.: 066461.11126 / 25WO1 CLY GATCAGAGTCATGAAAGAAGTCAA TCCACCAATTCATCCTGCCTCCATGA

[0074] Docket No.: 066461.11126 / 25WO1

[0075] Docket No.: 066461.11126 / 25WO1 NKG CCACCGCGCCCGGCCTAAAAATCTT AAAGAATAAATGAAAGAATAATTGTT

[0076] Docket No.: 066461.11126 / 25WO1

[0077] Docket No.: 066461.11126 / 25WO1 TRA ATCATGGCCTCTTGGCCAAGATTGA GGAAAAGTCCAAATAACTTCAGATTG

[0078] Docket No.: 066461.11126 / 25WO1

[0079] Docket No.: 066461.11126 / 25WO1 CD3 TCTGCCCCCTTAGGACACCTGCAGA CGATTCTTTTTGCGCGGCCTGAAATG

[0080] Docket No.: 066461.11126 / 25WO1

[0081] Docket No.: 066461.11126 / 25WO1 CD3 GGATGGAGAGAGGAAGTACCAAAT GCATAGTGATGTTTCCATACTAATAA

[0082] Docket No.: 066461.11126 / 25WO1

[0083] Docket No.: 066461.11126 / 25WO1 ' end IL2 CAACTCTGCACCAAGCGGACCTAA CAACAATTAACACCTTACCAACCAAT

[0084] Docket No.: 066461.11126 / 25WO1 ' end B2 CCTGGCCTCTTTTTTTTGTATATTAC GCTGACTTGACCCTTTGTCACCATATC

[0085] Docket No.: 066461.11126 / 25WO1 (SEQ ID NO: 103)

[0086] Docket No.: 066461.11126 / 25WO1 (2) Nucleic acids encoding an inactivated cell surface receptor In another general aspect, the present disclosure relates to an isolated nucleic acid encoding an inactivated cell surface receptor useful for embodiments of the application. It will be appreciated by those skilled in the art that the coding sequence of an inactivated cell surface receptor can be changed (e.g., replaced, deleted, inserted, etc.) without changing the amino acid sequence of the protein. Accordingly, it will be understood by those skilled in the art that nucleic acid sequences encoding an inactivated cell surface receptor of the application can be altered without changing the amino acid sequences of the proteins. In certain embodiments, an isolated nucleic acid encodes any inactivated cell surface receptor described herein, such as that comprises a monoclonal antibody-specific epitope, and / or a cytokine, such as an IL-15 or IL-2, wherein the monoclonal antibody- specific epitope and the cytokine are optionally operably linked by an autoprotease peptide sequence. In some embodiments, the isolated nucleic acid encodes an inactivated cell surface receptor comprising an epitope specifically recognized by an antibody, such as ibritumomab, tiuxetan, muromonab-CD3, tositumomab, abciximab, basiliximab, brentuximab vedotin, cetuximab, infliximab, rituximab, alemtuzumab, bevacizumab, certolizumab pegol, daclizumab, eculizumab, efalizumab, gemtuzumab, natalizumab, omalizumab, palivizumab, ranibizumab, tocilizumab, trastuzumab, vedolizumab, adalimumab, belimumab, canakinumab, denosumab, golimumab, ipilimumab, avelumab. ofatumumab, panitumumab, or ustekinumab. In some embodiments, the isolated nucleic acid encodes an inactivated cell surface receptor comprising an epitope specifically recognized by cetuximab. In some embodiments, the isolated nucleic acid encodes an inactivated cell surface receptor comprising an epitope specifically recognized by trastuzumab. In some embodiments, the isolated nucleic acid encodes an inactivated cell surface receptor comprising an epitope specifically recognized by bevacizumab. In some embodiments, the isolated nucleic acid encodes an inactivated cell surface receptor comprising an epitope specifically recognized by avelumab. In some embodiments, the isolated nucleic acid encodes an inactivated cell surface receptor comprising an epitope specifically recognized by ipilimumab. Docket No.: 066461.11126 / 25WO1 In certain embodiments, the isolated nucleic acid encodes an inactivated cell surface receptor having a truncated epithelial growth factor receptor (tEGFR) variant. Preferably, the inactivated cell surface receptor comprises an epitope specifically recognized by cetuximab, matuzumab, necitumumab or panitumumab, preferably cetuximab. In certain embodiments, the isolated nucleic acid encodes an inactivated cell surface receptor having one or more epitopes of CD79b, such as an epitope specifically recognized by polatuzumab vedotin. In certain embodiments, the isolated nucleic acid encodes an inactivated cell surface receptor having one or more epitopes of CD20, such as an epitope specifically recognized by rituximab. In certain embodiments, the isolated nucleic acid encodes an inactivated cell surface receptor having one or more epitopes of Her 2 receptor, such as an epitope specifically recognized by trastuzumab In certain embodiments, the autoprotease peptide sequence is porcine tesehovirus- 12A (P2A). In certain embodiments, the truncated epithelial growth factor receptor (tEGFR) variant consists of an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 71. In certain embodiments, the monoclonal antibody-specific epitope specifically recognized by polatuzumab vedotin consists of an amino acid sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, identical to SEQ ID NO: 78. In certain embodiments, the monoclonal antibody-specific epitope specifically recognized by rituximab consists of an amino acid sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, identical to SEQ ID NO: 80. In certain embodiments, the monoclonal antibody-specific epitope specifically recognized by trastuzumab consists of an amino acid sequence at least 90%, such as at Docket No.: 066461.11126 / 25WO1 least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%, identical to SEQ ID NO: 82. In certain embodiments, the IL-15 comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 72. In certain embodiments, the autoprotease peptide has an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 73. In certain embodiments, the polynucleotide sequence encodes a polypeptide comprising an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 74. In a particular embodiment, the isolated nucleic acid encoding the inactivated cell surface receptor comprises a polynucleotide sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100%, identical to SEQ ID NO: 75, preferably the polynucleotide sequence of SEQ ID NO: 75. In certain embodiments, the polynucleotide sequence encodes a polypeptide comprising an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 79. In another general aspect, the application provides a vector comprising a polynucleotide sequence encoding an inactivated cell surface receptor useful for embodiments of the application. Any vector known to those skilled in the art in view of the present disclosure can be used, such as a plasmid, a cosmid, a phage vector or a viral vector. In some embodiments, the vector is a recombinant expression vector such as a plasmid. The vector can include any element to establish a conventional function of an expression vector, for example, a promoter, ribosome binding element, terminator, enhancer, selection marker, and origin of replication. The promoter can be a constitutive, inducible, or repressible promoter. A number of expression vectors capable of delivering nucleic acids to a cell are known in the art and can be used herein for production of a inactivated cell surface receptor in the cell. Conventional cloning techniques or artificial Docket No.: 066461.11126 / 25WO1 gene synthesis can be used to generate a recombinant expression vector according to embodiments of the application. In a particular aspect, the application provides a vector for targeted integration of an inactivated cell surface receptor useful for embodiments of the application. In certain embodiments, the vector comprises an exogenous polynucleotide having, in the 5’ to 3’ order, (a) a promoter; (b) a polynucleotide sequence encoding an inactivated cell surface receptor, such as an inactivated cell surface receptor comprising a truncated epithelial growth factor receptor (tEGFR) variant and an interleukin 15 (IL-15), wherein the tEGFR variant and IL-15 are operably linked by an autoprotease peptide sequence, such as porcine tesehovirus-12A (P2A), and (c) a terminator / polyadenylation signal. In certain embodiments, the promoter is a CAG promoter. In certain embodiments, the CAG promoter comprises the polynucleotide sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100%, identical to SEQ ID NO: 63. Other promoters can also be used, examples of which include, but are not limited to, EF1a, UBC, CMV, SV40, PGK1, and human beta actin. In certain embodiments, the terminator / polyadenylation signal is a SV40 signal. In certain embodiments, the SV40 signal comprises the polynucleotide sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100%, identical to SEQ ID NO: 64. Other terminator sequences can also be used, examples of which include, but are not limited to BGH, hGH, and PGK. In certain embodiments, the polynucleotide sequence encoding an inactivated cell surface receptor comprises the polynucleotide sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100%, identical to SEQ ID NO: 75. In some embodiments, the vector further comprises a left homology arm and a right homology arm flanking the exogenous polynucleotide. In certain embodiments, the left homology arm comprises the polynucleotide sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100%, identical to SEQ ID NO: 130. In certain embodiments, the right homology arm comprises the polynucleotide sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100%, identical to SEQ ID NO: 131 Docket No.: 066461.11126 / 25WO1 (3) Nucleic acids encoding an HLA construct In another general aspect, the present disclosure relates to an isolated nucleic acid encoding an HLA construct useful for embodiments of the application. It will be appreciated by those skilled in the art that the coding sequence of an HLA construct can be changed (e.g., replaced, deleted, inserted, etc.) without changing the amino acid sequence of the protein. Accordingly, it will be understood by those skilled in the art that nucleic acid sequences encoding an HLA construct of the application can be altered without changing the amino acid sequences of the proteins. In certain embodiments, the isolated nucleic acid encodes an HLA construct comprising a signal peptide, such as an HLA-G signal peptide, operably linked to an HLA coding sequence, such as a coding sequence of a mature B2M, and / or a mature HLA-E. In some embodiments, the HLA coding sequence encodes the HLA-G and B2M, which are operably linked by a 4X GGGGS linker, and / or the B2M and HLA-E, which are operably linked by a 3X GGGGS linker. In a particular embodiment, the isolated nucleic acid encoding the HLA construct comprises a polynucleotide sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100%, identical to SEQ ID NO: 67, preferably the polynucleotide sequence of SEQ ID NO: 67. In another embodiment, the isolated nucleic acid encoding the HLA construct comprises a polynucleotide sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100%, identical to SEQ ID NO: 70, preferably the polynucleotide sequence of SEQ ID NO: 70. In another general aspect, the application provides a vector comprising a polynucleotide sequence encoding a HLA construct useful for embodiments of the application. Any vector known to those skilled in the art in view of the present disclosure can be used, such as a plasmid, a cosmid, a phage vector or a viral vector. In some embodiments, the vector is a recombinant expression vector such as a plasmid. The vector can include any element to establish a conventional function of an expression vector, for example, a promoter, ribosome binding element, terminator, enhancer, selection marker, and origin of replication. The promoter can be a constitutive, inducible, or repressible promoter. A number of expression vectors capable of delivering nucleic acids to a cell are known in the art and can be used herein for production of a HLA Docket No.: 066461.11126 / 25WO1 construct in the cell. Conventional cloning techniques or artificial gene synthesis can be used to generate a recombinant expression vector according to embodiments of the application. In a particular aspect, the application provides vectors for targeted integration of a HLA construct useful for embodiments of the application. In certain embodiments, the vector comprises an exogenous polynucleotide having, in the 5’ to 3’ order, (a) a promoter; (b) a polynucleotide sequence encoding an HLA construct; and (c) a terminator / polyadenylation signal. In certain embodiments, the promoter is a CAG promoter. In certain embodiments, the CAG promoter comprises the polynucleotide sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100%, identical to SEQ ID NO: 63. Other promoters can also be used, examples of which include, but are not limited to, EF1a, UBC, CMV, SV40, PGK1, and human beta actin. In certain embodiments, the terminator / polyadenylation signal is a SV40 signal. In certain embodiments, the SV40 signal comprises the polynucleotide sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100%, identical to SEQ ID NO: 64. Other terminator sequences can also be used, examples of which include, but are not limited to BGH, hGH, and PGK. In certain embodiments, a polynucleotide sequence encoding a HLA construct comprises a signal peptide, such as a HLA-G signal peptide, a mature B2M, and a mature HLA-E, wherein the HLA-G and B2M are operably linked by a 4X GGGGS linker (SEQ ID NO: 31) and the B2M transgene and HLA-E are operably linked by a 3X GGGGS linker (SEQ ID NO: 25). In particular embodiments, the HLA construct comprises the polynucleotide sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100%, identical to SEQ ID NO: 67, preferably the polynucleotide sequence of SEQ ID NO: 67. In another embodiment, the HLA construct comprises the polynucleotide sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100%, identical to SEQ ID NO: 70, preferably the polynucleotide sequence of SEQ ID NO: 70. In some embodiments, the vector further comprises a left homology arm and a right homology arm flanking the exogenous polynucleotide. Docket No.: 066461.11126 / 25WO1 In certain embodiments, the left homology arm comprises the polynucleotide sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100%, identical to SEQ ID NO: 132. In certain embodiments, the right homology arm comprises the polynucleotide sequence at least 90%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 100%, identical to SEQ ID NO: 133. (4) Host cells In another general aspect, the application provides a host cell comprising a vector of the application and / or an isolated nucleic acid encoding a construct of the application. Any host cell known to those skilled in the art in view of the present disclosure can be used for recombinant expression of exogenous polynucleotides of the application. According to particular embodiments, the recombinant expression vector is transformed into host cells by conventional methods such as chemical transfection, heat shock, or electroporation, where it is stably integrated into the host cell genome such that the recombinant nucleic acid is effectively expressed. Examples of host cells include, for example, recombinant cells containing a vector or isolated nucleic acid of the application useful for the production of a vector or construct of interest; or an engineered iPSC or derivative cell thereof containing one or more isolated nucleic acids of the application, preferably integrated at one or more chromosomal loci. A host cell of an isolated nucleic acid of the application can also be an immune effector cell, such as a T cell or NK cell, comprising the one or more isolated nucleic acids of the application. The immune effector cell can be obtained by differentiation of an engineered iPSC of the application. Any suitable method in the art can be used for the differentiation in view of the present disclosure. The immune effector cell can also be obtained transfecting an immune effector cell with one or more isolated nucleic acids of the application. Cell Compositions In another general aspect, the application provides a composition comprising an isolated polynucleotide of the application, a host cell and / or an iPSC or derivative cell thereof of the application. Docket No.: 066461.11126 / 25WO1 In certain embodiments, the composition further comprises one or more therapeutic agents selected from the group consisting of a peptide, a cytokine, a checkpoint inhibitor, a mitogen, a growth factor, a small RNA, a dsRNA (double stranded RNA), siRNA, oligonucleotide, mononuclear blood cells, , a vector comprising one or more polynucleic acids of interest, an antibody, a chemotherapeutic agent or a radioactive moiety, or an immunomodulatory drug (IMiD). In certain embodiments, the composition is a pharmaceutical composition comprising an isolated polynucleotide of the application, a host cell and / or an iPSC or derivative cell thereof of the application and a pharmaceutically acceptable carrier. The term “pharmaceutical composition” as used herein means a product comprising an isolated polynucleotide of the application, an isolated polypeptide of the application, a host cell of the application, and / or an iPSC or derivative cell thereof of the application together with a pharmaceutically acceptable carrier. Polynucleotides, polypeptides, host cells, and / or iPSCs or derivative cells thereof of the application and compositions comprising them are also useful in the manufacture of a medicament for therapeutic applications mentioned herein. As used herein, the term “carrier” refers to any excipient, diluent, filler, salt, buffer, stabilizer, solubilizer, oil, lipid, lipid containing vesicle, microsphere, liposomal encapsulation, or other material well known in the art for use in pharmaceutical formulations. It will be understood that the characteristics of the carrier, excipient or diluent will depend on the route of administration for a particular application. As used herein, the term “pharmaceutically acceptable carrier” refers to a non-toxic material that does not interfere with the effectiveness of a composition described herein or the biological activity of a composition described herein. According to particular embodiments, in view of the present disclosure, any pharmaceutically acceptable carrier suitable for use in a polynucleotide, polypeptide, host cell, and / or iPSC or derivative cell thereof can be used. The formulation of pharmaceutically active ingredients with pharmaceutically acceptable carriers is known in the art, e.g., Remington: The Science and Practice of Pharmacy (e.g.21st edition (2005), and any later editions). Non-limiting examples of additional ingredients include: buffers, diluents, solvents, tonicity regulating agents, Docket No.: 066461.11126 / 25WO1 preservatives, stabilizers, and chelating agents. One or more pharmaceutically acceptable carrier may be used in formulating the pharmaceutical compositions of the application. Methods of Use of Engineered Cells Primary cancer cells can be readily distinguished from non-cancerous cells by well-established techniques, particularly histological examination. The definition of a cancer cell, as used herein, includes not only a primary cancer cell, but any cell derived from a cancer cell ancestor. This includes metastasized cancer cells, and in vitro cultures and cell lines derived from cancer cells. When referring to a type of cancer that normally manifests as a solid tumour, a "clinically detectable" tumour is one that is detectable on the basis of tumour mass; e.g., by procedures such as computed tomography (CT) scan, magnetic resonance imaging (MRI), X-ray, ultrasound or palpation on physical examination, and / or which is detectable because of the expression of one or more cancer- specific antigens in a sample obtainable from a patient. Cancer conditions may be characterized by the abnormal proliferation of malignant cancer cells and may include leukemias, such as AML, CML, ALL and CLL, lymphomas, such as Hodgkin lymphoma, non-Hodgkin lymphoma and multiple myeloma, and solid cancers such as sarcomas, skin cancer, melanoma, bladder cancer, brain cancer, breast cancer, uterus cancer, ovary cancer, prostate cancer, lung cancer, colorectal cancer, cervical cancer, liver cancer, head and neck cancer, esophageal cancer, pancreatic cancer, renal cancer, adrenal cancer, stomach cancer, testicular cancer, cancer of the gall bladder and biliary tracts, thyroid cancer, thymus cancer, cancer of bone, and cerebral cancer, as well as cancer of unknown primary (CUP). Cancer cells within an individual may be immunologically distinct from normal somatic cells in the individual (i.e. the cancerous tumour may be immunogenic). For example, the cancer cells may be capable of eliciting a systemic immune response in the individual against one or more antigens expressed by the cancer cells. The tumour antigens that elicit the immune response may be specific to cancer cells or may be shared by one or more normal cells in the individual. Docket No.: 066461.11126 / 25WO1 The cancer cells of an individual suitable for treatment as described herein may express the antigen and / or may be of correct HLA type to bind the antigen receptor expressed by the Τ cells. An individual suitable for treatment as described above may be a mammal. In preferred embodiments, the individual is a human. In other preferred embodiments, non- human mammals, especially mammals that are conventionally used as models for demonstrating therapeutic efficacy in humans (e.g. murine, primate, porcine, canine, or rabbit animals) may be employed. In some embodiments, the individual may have minimal residual disease (MRD) after an initial cancer treatment. In some embodiments, the individual may have no minimal residual disease after one or more cancer treatments or repeated dosing. In some aspects, the present disclosure provides methods for treating a subject with a therapeutic dose of a composition comprising iPSCs or a derivative cell thereof (e.g., a genetically engineered allogenic NK or T cell). In some embodiments, such methods can comprise a step of administering a therapeutically effective dose of the composition. In some cases, a single therapeutically effective dose is administered, In some cases, two or more therapeutically effective doses are administered in series. In some embodiments, engineered cells of the present disclosure may be administered in a single dose, and such administration may be by injection, e.g., intravenous injection. In some embodiments, engineered cell of the present disclosure may be administered in multiple doses. Dosing may be once, twice, three times, four times, five times, six times, or more than six times per year. Dosing may be once a month, once every two weeks, once a week, or once every other day. Administration of engineered cells of the present disclosure may continue as long as necessary. Suitable administration of a therapeutically effective doses can comprise administration of a single dose, or can comprise administration of sequential doses daily, semi-weekly, weekly, once every two weeks, once a month, annually, etc. In some cases, a therapeutically effective dose is administered as two or more doses of escalating concentration (i.e., increasing doses), where (i) all of the doses are therapeutic doses, or where (ii) a sub-therapeutic dose (or two or more sub-therapeutic doses) is initially given and therapeutic doses are achieved by said escalation. As one non-limiting example to illustrate escalating concentration Docket No.: 066461.11126 / 25WO1 (i.e., increasing doses), a therapeutically effective dose can be administered weekly, beginning with a sub-therapeutic dose, and each subsequent dose can be increased by a particular increment, or by variable increments, until a therapeutic dose is reached, at which point administration may cease or may continue. As another non-limiting example to illustrate escalating concentration, a therapeutically effective dose can be administered weekly, beginning with a therapeutic dose, and each subsequent dose can be increased by a particular increment, or by variable increments, until a therapeutic dose is reached, at which point administration may cease or may continue. In some embodiments, administration of a therapeutically effective dose can be a continuous infusion and the dose can altered (e.g., escalated) over time. Dosage and frequency may vary depending on the half-life of the cell composition in the patient, and / or mode of administration. An individual with cancer may display at least one identifiable sign, symptom, or laboratory finding that is sufficient to make a diagnosis of cancer in accordance with clinical standards known in the art. Examples of such clinical standards can be found in textbooks of medicine such as Harrison’s Principles of Internal Medicine, 15th Ed., Fauci AS et al., eds., McGraw-Hill, New York, 2001. In some instances, a diagnosis of a cancer in an individual may include identification of a particular cell type (e.g. a cancer cell) in a sample of a body fluid or tissue obtained from the individual. An anti-tumor effect is a biological effect which can be manifested by a reduction in the rate of tumor growth, decrease in tumor volume, a decrease in the number of tumor cells, a decrease in the number of metastases, an increase in life expectancy, or amelioration of various physiological symptoms associated with the cancerous condition. An "anti-tumor effect" can also be manifested by the ability of the peptides, polynucleotides, cells and antibodies, also Τ cells which may be obtained according to the methods of the present disclosure , as described herein in prevention of the occurrence of tumors in the first place. Treatment may be any treatment and / or therapy, whether of a human or an animal (e.g. in veterinary applications), in which some desired therapeutic effect is achieved, for example, the inhibition or delay of the progress of the condition, and includes a reduction in the rate of progress, a halt in the rate of progress, amelioration of the condition, cure or remission (whether partial or total) of the condition, preventing, delaying, abating or Docket No.: 066461.11126 / 25WO1 arresting one or more symptoms and / or signs of the condition or prolonging survival of a subject or patient beyond that expected in the absence of treatment. Treatment may also be prophylactic (i.e. prophylaxis). For example, an individual susceptible to or at risk of the occurrence or re-occurrence of cancer may be treated as described herein. Such treatment may prevent or delay the occurrence or re-occurrence of cancer in the individual. In particular, treatment may include inhibiting cancer growth, including complete cancer remission, and / or inhibiting cancer metastasis. Cancer growth generally refers to any one of a number of indices that indicate change within the cancer to a more developed form. Thus, indices for measuring an inhibition of cancer growth include a decrease in cancer cell survival, a decrease in tumor volume or morphology (for example, as determined using computed tomographic (CT), sonography, or other imaging method), a delayed tumor growth, a destruction of tumor vasculature, improved performance in delayed hypersensitivity skin test, an increase in the activity of Τ cells, and a decrease in levels of tumor-specific antigens. Administration of Τ cells modified as described herein may improve the capacity of the individual to resist cancer growth, in particular growth of a cancer already present the subject and / or decrease the propensity for cancer growth in the individual. This application provides a method of treating a disease or a condition in a subject in need thereof. The methods comprise administering to the subject in need thereof a therapeutically effective amount of cells of the application and / or a composition of the application. In certain embodiments, the disease or condition is cancer. The cancer can, for example, be a solid or a liquid cancer. The cancer, can, for example, be selected from the group consisting of a lung cancer, a gastric cancer, a colon cancer, a liver cancer, a renal cell carcinoma, a bladder urothelial carcinoma, a metastatic melanoma, a breast cancer, an ovarian cancer, a cervical cancer, a head and neck cancer, a pancreatic cancer, an endometrial cancer, a prostate cancer, a thyroid cancer, a glioma, a glioblastoma, and other solid tumors, and a non-Hodgkin’s lymphoma (NHL), Hodgkin’s lymphoma / disease (HD), an acute lymphocytic leukemia (ALL), a chronic lymphocytic leukemia (CLL), a chronic myelogenous leukemia (CML), a multiple myeloma (MM), an Docket No.: 066461.11126 / 25WO1 acute myeloid leukemia (AML), and other liquid tumors. In a preferred embodiment, the cancer is a non-Hodgkin’s lymphoma (NHL). According to embodiments of the application, the composition comprises a therapeutically effective amount of an isolated polynucleotide, an isolated polypeptide, a host cell, and / or an iPSC or derivative cell thereof. As used herein, the term “therapeutically effective amount” refers to an amount of an active ingredient or component that elicits the desired biological or medicinal response in a subject. A therapeutically effective amount can be determined empirically and in a routine manner, in relation to the stated purpose. As used herein with reference to a cell of the application and / or a pharmaceutical composition of the application a therapeutically effective amount means an amount of the cells and / or the pharmaceutical composition that modulates an immune response in a subject in need thereof. According to particular embodiments, a therapeutically effective amount refers to the amount of therapy which is sufficient to achieve one, two, three, four, or more of the following effects: (i) reduce or ameliorate the severity of the disease, disorder or condition to be treated or a symptom associated therewith; (ii) reduce the duration of the disease, disorder or condition to be treated, or a symptom associated therewith; (iii) prevent the progression of the disease, disorder or condition to be treated, or a symptom associated therewith; (iv) cause regression of the disease, disorder or condition to be treated, or a symptom associated therewith; (v) prevent the development or onset of the disease, disorder or condition to be treated, or a symptom associated therewith; (vi) prevent the recurrence of the disease, disorder or condition to be treated, or a symptom associated therewith; (vii) reduce hospitalization of a subject having the disease, disorder or condition to be treated, or a symptom associated therewith; (viii) reduce hospitalization length of a subject having the disease, disorder or condition to be treated, or a symptom associated therewith; (ix) increase the survival of a subject with the disease, disorder or condition to be treated, or a symptom associated therewith; (xi) inhibit or reduce the disease, disorder or condition to be treated, or a symptom associated therewith in a subject; and / or (xii) enhance or improve the prophylactic or therapeutic effect(s) of another therapy. Docket No.: 066461.11126 / 25WO1 The therapeutically effective amount or dosage can vary according to various factors, such as the disease, disorder or condition to be treated, the means of administration, the target site, the physiological state of the subject (including, e.g., age, body weight, health), whether the subject is a human or an animal, other medications administered, and whether the treatment is prophylactic or therapeutic. Treatment dosages are optimally titrated to optimize safety and efficacy. According to particular embodiments, the compositions described herein are formulated to be suitable for the intended route of administration to a subject. For example, the compositions described herein can be formulated to be suitable for intravenous, subcutaneous, or intramuscular administration. The cells of the application and / or the pharmaceutical compositions of the application can be administered in any convenient manner known to those skilled in the art. For example, the cells of the application can be administered to the subject by aerosol inhalation, injection, ingestion, transfusion, implantation, and / or transplantation. The compositions comprising the cells of the application can be administered transarterially, subcutaneously, intradermaly, intratumorally, intranodally, intramedullary, intramuscularly, intrapleurally, by intravenous (i.v.) injection, or intraperitoneally. In certain embodiments, the cells of the application can be administered with or without lymphodepletion of the subject. The pharmaceutical compositions comprising cells of the application can be provided in sterile liquid preparations, typically isotonic aqueous solutions with cell suspensions, or optionally as emulsions, dispersions, or the like, which are typically buffered to a selected pH. The compositions can comprise carriers, for example, water, saline, phosphate buffered saline, and the like, suitable for the integrity and viability of the cells, and for administration of a cell composition. Sterile injectable solutions can be prepared by incorporating cells of the application in a suitable amount of the appropriate solvent with various other ingredients, as desired. Such compositions can include a pharmaceutically acceptable carrier, diluent, or excipient such as sterile water, physiological saline, glucose, dextrose, or the like, that are suitable for use with a cell composition and for administration to a subject, such as a human. Suitable buffers for providing a cell composition are well known in the art. Any Docket No.: 066461.11126 / 25WO1 vehicle, diluent, or additive used is compatible with preserving the integrity and viability of the cells of the application. The cells of the application and / or the pharmaceutical compositions of the application can be administered in any physiologically acceptable vehicle. A cell population comprising cells of the application can comprise a purified population of cells. Those skilled in the art can readily determine the cells in a cell population using various well known methods. The ranges in purity in cell populations comprising genetically modified cells of the application can be from about 50% to about 55%, from about 55% to about 60%, from about 60% to about 65%, from about 65% to about 70%, from about 70% to about 75%, from about 75% to about 80%, from about 80% to about 85%, from about 85% to about 90%, from about 90% to about 95%, or from about 95% to about 100%. Dosages can be readily adjusted by those skilled in the art, for example, a decrease in purity could require an increase in dosage. The cells of the application are generally administered as a dose based on cells per kilogram (cells / kg) of body weight of the subject to which the cells and / or pharmaceutical compositions comprising the cells are administered. Generally, the cell doses are in the range of about 104to about 1010cells / kg of body weight, for example, about 105to about 109, about 105to about 108, about 105to about 107, or about 105to about 106, depending on the mode and location of administration. In general, in the case of systemic administration, a higher dose is used than in regional administration, where the immune cells of the application are administered in the region of a tumor and / or cancer. Exemplary dose ranges include, but are not limited to, 1 x 104to 1 x 108, 2 x 104to 1 x 108, 3 x 104to 1 x 108, 4 x 104to 1 x 108, 5 x 104to 6 x 108, 7 x 104to 1 x 108, 8 x 104to 1 x 108, 9 x 104to 1 x 108, 1 x 105to 1 x 108, 1 x 105to 9 x 107, 1 x 105to 8 x 107, 1 x 105to 7 x 107, 1 x 105to 6 x 107, 1 x 105to 5 x 107, 1 x 105to 4 x 107, 1 x 105to 4 x 107, 1 x 105to 3 x 107, 1 x 105to 2 x 107, 1 x 105to 1 x 107, 1 x 105to 9 x 106, 1 x 105to 8 x 106, 1 x 105to 7 x 106, 1 x 105to 6 x 106, 1 x 105to 5 x 106, 1 x 105to 4 x 106, 1 x 105to 4 x 106, 1 x 105to 3 x 106, 1 x 105to 2 x 106, 1 x 105to 1 x 106, 2 x 105to 9 x 107, 2 x 105to 8 x 107, 2 x 105to 7 x 107, 2 x 105to 6 x 107, 2 x 105to 5 x 107, 2 x 105to 4 x 107, 2 x 105to 4 x 107, 2 x 105to 3 x 107, 2 x 105to 2 x 107, 2 x 105to 1 x 107, 2 x 105to 9 x 106, 2 x 105to 8 x 106, 2 x 105to 7 x 106, 2 x 105to 6 x 106, 2 x 105to 5 x 106, 2 x 105to Docket No.: 066461.11126 / 25WO1 4 x 106, 2 x 105to 4 x 106, 2 x 105to 3 x 106, 2 x 105to 2 x 106, 2 x 105to 1 x 106, 3 x 105to 3 x 106cells / kg, and the like. Additionally, the dose can be adjusted to account for whether a single dose is being administered or whether multiple doses are being administered. The precise determination of what would be considered an effective dose can be based on factors individual to each subject. As used herein, the terms “treat,” “treating,” and “treatment” are all intended to refer to an amelioration or reversal of at least one measurable physical parameter related to a cancer, which is not necessarily discernible in the subject, but can be discernible in the subject. The terms “treat,” “treating,” and “treatment,” can also refer to causing regression, preventing the progression, or at least slowing down the progression of the disease, disorder, or condition. In a particular embodiment, “treat,” “treating,” and “treatment” refer to an alleviation, prevention of the development or onset, or reduction in the duration of one or more symptoms associated with the disease, disorder, or condition, such as a tumor or more preferably a cancer. In a particular embodiment, “treat,” “treating,” and “treatment” refer to prevention of the recurrence of the disease, disorder, or condition. In a particular embodiment, “treat,” “treating,” and “treatment” refer to an increase in the survival of a subject having the disease, disorder, or condition. In a particular embodiment, “treat,” “treating,” and “treatment” refer to elimination of the disease, disorder, or condition in the subject. The cells of the application and / or the pharmaceutical compositions of the application can be administered in combination with one or more additional therapeutic agents. In certain embodiments the one or more therapeutic agents are selected from the group consisting of a peptide, a cytokine, a checkpoint inhibitor, a mitogen, a growth factor, a small RNA, a dsRNA (double stranded RNA), siRNA, oligonucleotide, mononuclear blood cells, a vector comprising one or more polynucleic acids of interest, an antibody, a chemotherapeutic agent or a radioactive moiety, or an immunomodulatory drug (IMiD). In certain embodiments, the one or more therapeutic agents comprise an antibody. In certain embodiments, the one or more therapeutic agents comprise one or more antibodies independently selected from the group consisting of ibritumomab, tiuxetan, muromonab-CD3, tositumomab, abciximab, basiliximab, brentuximab vedotin, cetuximab, infliximab, rituximab, alemtuzumab, bevacizumab, certolizumab pegol, Docket No.: 066461.11126 / 25WO1 daclizumab, eculizumab, efalizumab, gemtuzumab, natalizumab, omalizumab, palivizumab, polatuzumab vedotin, ranibizumab, tocilizumab, trastuzumab, vedolizumab, adalimumab, belimumab, canakinumab, denosumab, golimumab, ipilimumab, avelumab, ofatumumab, panitumumab, and ustekinumab. In certain embodiments, the one or more therapeutic agents comprise cetuximab. In certain embodiments, the one or more therapeutic agents comprise trastuzumab. In certain embodiments, the one or more therapeutic agents comprise bevacizumab. In certain embodiments, the one or more therapeutic agents comprise avelumab. In certain embodiments, the one or more therapeutic agents comprise ipilimumab. EXAMPLES Abbreviations E:T Effector to target ratio uL Microliter mM Millimolar x g Times gravity oC Degrees Celsius ug Microgram UTD Untransduced HSA Human serum albumin h or hr Hour WT Wild type Peripheral blood Min Minute PBMC mononuclear cells RPM Rounds per minute iNK iPSC-derived NK Cell ICL Intracellular loop iT iPSC-derived T cell GPCR G protein-coupled receptor mL Milliliter CAR Chimeric antigen receptor Example 1. Expression of GPCR CARs in Nurkat Cells Rationale Conventional CARs have a single transmembrane domain, and are thus limited in their engineerability because additional extracellular or intracellular domains must be engineered onto the receptor in a linear fashion. For extracellular domains, additional binders to increase CAR valency or specificity for one or more antigens can reduce functionality by increasing the distance between cells in an immune synapse. For Docket No.: 066461.11126 / 25WO1 intracellular domains, additional domains can increase the distance of those domains from membrane bound kinases involved in signal transduction (e.g., Lck) and hamper their functionality. Typically, these issues have been overcome by engineering multiple single pass receptors. A multi-transmembrane domain-containing CAR can allow a single receptor to contain multiple binders or additional endodomains without a reduction in functionality or diminished signal transduction. Experimental Design Receptors were designed that included fusion of the 9G8 anti-EGFR binder to the N terminus of CXCR3 to confer antigen binding capabilities, and used as a template to incorporate immunostimulatory endodomains, including CD3z, 4-1BB, CD28, and OX40. The schema for engineering of these receptors is shown in Table 8 and FIG.1A- H. Mutation set refers to whether the CXCR3 sequence was mutated or kept as the wild- type AA sequence. WT = wild-type. C / N = ecto / endo dead containing mutations according to Colvin et al, 2006 (10.1128 / MCB.00556-06), which is incorporated by reference herein in its entirety, to eliminate binding of CXCR3 to its cognate chemokines and abrogate CXCR3-mediated chemotaxis. Those relevant mutated residues are Y27A, Y29A, R149N, S245A, and deletion of L332 - L368. ICL1 / 2 / 3 = intracellular loop 1 / 2 / 3. 28 = CD28 costimulatory domain. BB = 4-1BB costimulatory domain. G4S-28, CD28 costimulatory domain with a preceding GGGGS linker. OX40 = OX40 costimulatory domain. z = CD3z activating domain. x = no endomain inserted. Table 8. Construct Mutation N- ICL1 ICL2 ICL3 C- s Docket No.: 066461.11126 / 25WO1 AMD228 WT 9G8 28 x x z AMD229 WT 9G8 G4S- x x z atera s ^ NEB Hifi DNA Assembly Master Mix (NEB E2621X) ^ NEB XhoI (NEB R0146M) ^ NEB BamHI-HF (NEB R3136M) ^ Primary human PBMCs (cryopreserved, from Hemacare) ^ Nurkat cells (Nur77-GFP Jurkat reporter line) ^ Polybrene 10mg / mL (EMD Millipore CAT# TR-1003-G) ^ Pan T cell isolation kit (Miltenyi CAT#130-096-535), with associated Miltenyi MACS buffer + BSA ^ T Cell Transact (Miltenyi CAT#130-111-160) ^ Transdux MAX (SystemBio LV860A-1) ^ Recombinant human IL-2 (IL2 info: Miltenyi IL-2 (Lot# 5210307089) ; prepared by Sam M at 3711 Market; 1mg / mL, which amounts to 1.6E7 UNITS / mL) ^ R10 media (RPMI 1640 Medium (Gibco CAT# 21875034) with 10% Fetal Bovine Serum (Gibco CAT# 10438026)) ^ Cell Trace Violet cell stain (Thermofisher CAT# C34557) Docket No.: 066461.11126 / 25WO1 ^ Nalm6-NLR, Nalm6-EGFR-NLR cells ^ LIVE / DEAD™ Fixable Near-IR Dead Cell Stain Kit (Thermofisher CAT# L10119) ^ BD Staining Buffer, BSA (BD Pharmingen, Cat# 554657) ^ MonoRab Rabbit anti-Camelid VHH Cocktail, AF647 conjugated (Genscript, CAT#A02014) Expression of GPCR-CAR Constructs CXCR3 GPCR CAR constructs were packaged into lentivirus and transduced into Nurkat cells. Nurkats were resuspended in R10 media +3ug / mL polybrene at .5e6 / mL and plated 500uL per well into 24 well plates.100uL lentivirus was applied to each well. p2320 (9G8-CD8HTM.BB.z) and p2321 (9G8-CD28HTMcostim.z) were used as conventional CAR controls.3 days post transduction, CAR expression was assessed. 100uL cell suspension was plated at n=2 in a 96 well v-bottom plate. Plates were washed twice, and centrifuged for 3min at 300xg. Supernatant was discarded and 150uL BD stain buffer was added, followed by brief shaking (~2400rpm) on the vortexer to resuspend the cell pellet.50uL of primary stain containing anti-Camelid VHH AF647 (Genscript) 1:500 was added, and cells were incubated at 4oC for 25-30min. Cells were washed twice as before and resuspended in 30uL stain buffer for Intellicyte analysis. Flow cytometry data was collected using the Intellicyte cytometer and analyzed in FlowJo. Results As shown in Fig.2A, the flow cytometry gating strategy for GPCR-CAR- expressing nurkat cells. RL1-A is the area channel for fluorescence from Alexa Fluor 647. As shown in Fig.2B, Most GPCR CAR Constructs yield expression greater than 50%. Constructs in which CD28 costimulatory domain was inserted into ICL1 (AMD204, 205, 222) show reduced expression. Conclusions Docket No.: 066461.11126 / 25WO1 GPCR CARs showed high expression levels greater than 50%, and most constructs were comparable in expression level to the conventional CAR controls in the sample set, p2320 and p2321. This demonstrated the feasibility of engineering the intracellular loops of GPCRs to incorporate additional domains. The reduced expression of ICL1 insertion GPCR CARs demonstrated that certain engineering strategies can yield more stable and better-expressing receptors. Example 2. Expression of GPCR CARs in Primary T Cells Donor PBMCs were thawed (>90%% viability post thaw, ~75e6 cells total). T cells were isolated from the PBMCs using the Miltenyi T cell isolation kit according to the manufacturer's protocol. Final T cell yield was ~12e6 T cells at >98%% viability. Cell concentration was adjusted to 1e6 in R10+50U / mL human IL2, and 2mL was pipetted into 4 wells each of a 24 well plate (8e6 total), 20uL Miltenyi TransAct reagent was added to each well to activate the T cells. Cells were incubated at 37oC for two days. Activated T cells were combined and counted (~8e6 total cells).5.25e6 cells were pelleted. A mastermix of transduction-cell suspension was created according to Table 9. Table 9. Transduction Reagent volumes for (20+1 Reactions) R10+50U IL2 / mL Media (mL) 84 500uL of mastermix was plated into each well of 20 wells of a 24 well plate.  100uL of lentivirus containing the blow plasmids was added to designated wells. The 20th well was untransduced (UTD) and received no virus. T cells were expanded as needed every other or every 3rd day to maintain a suspension of ~.5e6 cells / mL without exceeding 2e6 / mL.7 days post transduction, CAR expression was assessed.100uL cell suspension was plated at n=2 in a 96 well v-bottom plate. Plates were washed twice, and centrifuged for 3min at 300xg. Supernatant was dumped and 150uL BD stain buffer was added, followed by brief shaking (~2400rpm) on the vortexer to resuspend the cell pellet. 50uL primary stain containing anti-Camelid VHH AF647 (Genscript) 1:500 and Sytox Docket No.: 066461.11126 / 25WO1 Green (Invitrogen) 1:1000 was added. Cells were incubated at 4oC for 25-30min. Cells were washed twice as before and resuspended in 30uL stain buffer for Intellicyte analysis. Flow cytometry data was collected using the Intellicyte cytometer and analyzed in FlowJo. Results As shown in Fig.3A, flow cytometry gating strategy for detection of GPCR-CAR transduced primary T cells. BL2-A is the area channel for fluorescence of Sytox Green Dead cell stain. RL1-A is the area channel for fluorescence of AF647 anti-VHH cocktail for detection of VHH-containing CARs. Gating strategy shown for untransduced (UTD) T cells. As shown in Fig.3B, CAR expression values of transduced primary T cells as a percentage of live cells. Conclusions CXCR3 GPCR CARs having the C / N mutation set showed poor expression (~<10% CAR+). WT CXCR3 CARs had a range of expression, with AMD200, AMD208, and AMD232 having relatively high expression levels, comparable to or higher than the conventional CAR controls. AMD246, which incorporates CD28, OX40, 4-1BB and CD3z into one receptor also had relatively high expression. Incorporation of a CD28 costimulatory domain reduced CAR expression. For example, AMD 204 and AMD240 both have insertions in ICL1, however AMD204, which includes a CD28 insertion into ICL1, expressed significantly worse than AMD240, which incorporates OX40 into ICL1.9G8-CXCR3 constructs of both the WT and C / N varieties had high expression near or greater than 50% positive, suggesting that incorporation of additional endodomains disrupted expression of C / N constructs. Example 3. T Cell Cytotoxicity Against Nalm6 Cells WT CXCR3 CAR constructs were advanced for a cytotoxicity analysis against Nalm6 NLR and Nalm6 NLR EGFR cells. Nalm6 NLR without EGFR are a negative control and test whether the GPCR CAR format may create nonspecific cytotoxicity. Docket No.: 066461.11126 / 25WO1 Each CAR T was cocultured with both target cell lines at 2:1, 1:1, 1:2, 1:5, and 1:10 E:T ratios, adjusted for CAR%. Since CAR positivity was variable among the CAR Ts tested, several UTD T cell cocultures were prepared with the amount of UTD T cells added adjusted as though they had 10%, 20%, 30%, or 45% CAR expression.20e6 of each target cell line was labeled with CTV according to the manufacturer’s suggested protocol and resuspended @ .4e6 / mL.75uL of target cell line was added to each well according to the plate map for .03e6 target cells / well. T cells were counted, aliquoted, pelleted, and then resuspended according to Table 10. The resuspension volume for each T cell was determined based on the cell concentration needed for a 2:1 E:T ratio. This stock was then diluted to make cell suspensions for the 1:1, 1:2, 1:5, and 1:10 E:T ratios.

[0087] Docket No.: 066461.11126 / 25WO1 Table 10. AM ID 20 20 20 20 22 23 24 24 9G CXC p23 p23 3G-9 UT 10 UT 20 UT 30 UT 45%. . . . . . .

[0088] Docket No.: 066461.11126 / 25WO1 For each plate, cocultures were mixed and then transferred to 96 well V bottom plates. Plates were washed twice. Plates were centrifuged for 3min at 300xg. Supernatant was discarded and 150uL BD stain buffer was added, followed by brief shaking (~2400rpm) on the vortexer to resuspend the cell pellet.50uL primary stain containing Near IR Fluorescent Reactive Dye (Invitrogen) 1:1000 and anti-human CD25 PE (Biolegend) 1:50 was added. Cells were incubated at 4oC for 25-30min. Cells were washed twice as before and resuspended in 30uL stain buffer for Intellicyte analysis. Flow cytometry data was collected using the Intellicyte cytometer and analyzed in FlowJo. Results As shown in Fig.5, cytotoxicity of GPCR-CAR expressing T cells compared to T cells transduced with conventional CAR equivalents. T cell samples are grouped by similar CARs. CARs with 4-1BB and CD3z only are grouped, with p2320 being the conventional CAR equivalent. CD28- and CD3z-only CARs are grouped, with p2321 being the conventional CAR equivalent. Finally, CARs with CD28, 4-1BB, (and for AMD246, also OX40) and CD3z are grouped together, with 3G-9G8 being a CD28 / 4- 1BB / CD3z 3rdgeneration conventional CAR equivalent. Conclusions Most GPCR CAR constructs conferred cytotoxicity nearly equivalent to or surpassing their conventional CAR counterparts. The exceptions to this were AMD204, and AMD246, which both incorporate costimulatory domains into ICL1. This engineering step reduced functionality of the receptors, possibly due to reduced expression levels. Constructs with CD28 inserted into ICL3, AMD232 and AMD208 showed the highest levels of cytotoxicity in the assay. It will be appreciated by those skilled in the art that changes could be made to the embodiments described above without departing from the broad inventive concept thereof. It is understood, therefore, that this invention is not limited to the particular Docket No.: 066461.11126 / 25WO1 embodiments disclosed, but it is intended to cover modifications within the spirit and scope of the present invention as defined by the present description.

Claims

Docket No.: 066461.11126 / 25WO1 CLAIMS It is claimed:

1. A primary cell, an induced pluripotent stem cell (iPSC) or a derivative cell thereof comprising: at least one exogenous polynucleotide encoding one or more chimeric antigen receptors (CARs), said one or more CARs comprising: (i) a G protein-coupled receptor (GPCR) scaffold or fragment thereof, and (ii) an antigen binding domain targeting at least one tumor antigen, and (iii) at least one intracellular domain; and optionally, at least one of: (i) a deletion or reduced expression of one or more of B2M, TAP 1, TAP 2, Tapasin, RFXANK, CIITA, RFX5, RFXAP genes, (ii) an exogenous polynucleotide encoding a human leukocyte antigen E (HLA-E) and / or human leukocyte antigen G (HLA-G), (iii) an exogenous polynucleotide encoding a natural killer (NK) cell receptor immunoglobulin gamma Fc region receptor III (FcyRIII, cluster of differentiation 16 (CD16)) and / or an NKG2D protein, (iv) a deletion or reduced expression of one or more of NKG2A CD70, CD38 or CD33 genes, (v) an exogeneous polynucleotide encoding a cytokine, (vi) an exogenous polynucleotide encoding a safety switch, and (vii) an exogeneous polynucleotide encoding a PSMA cell tracer (viii) an exogeneous polynucleotide encoding a membrane bound IL-12 polypeptide.

2. The primary cell, the iPSC or the derivative cell according to claim 1, wherein the GPCR scaffold or fragment thereof is selected from the group consisting of rhodopsin-like receptors (Class A), secretin family receptors (Class B), or Metabotropic receptors (Class C).Docket No.: 066461.11126 / 25WO1 3. The primary cell, the iPSC or the derivative cell according to claim 2 comprising the rhodopsin-like receptors (Class A), wherein the GPCR scaffold is a chemokine receptor selected from the group consisting of CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CCR10, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CXCR6, CX3CR1, XCR1, ACKR1, ACKR2, ACKR3, ACKR4, and a fragment of any receptor thereof.

4. The primary cell, the iPSC or the derivative cell according to claim 3, wherein the GPCR scaffold is CXCR3 or a fragment thereof.

5. The primary cell, the iPSC or the derivative cell according to claim 4, wherein (i) the CXCR3 comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 134, or (ii) the CXCR3 is encoded by a polynucleotide having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one or more of SEQ ID NOs: 135-151.

6. The primary cell, the iPSC or the derivative cell according to any one of claims 1- 5, wherein the GPCR scaffold comprises: (i) at least one an extracellular domain comprising the first antigen- binding domain, said extracellular domain comprising an extracellular loop of the GPCR scaffold; (ii) at least one transmembrane domain; and (iii) and the at least one intracellular domain.

7. The primary cell, the iPSC or the derivative cell according to claim 6, wherein the at least one intracellular domain comprises: (i) one or more costimulatory signaling domains selected from the group consisting of CD28, 41BB, IL2Rb, CD40, OX40 (CD134), CD80,Docket No.: 066461.11126 / 25WO1 CD86, CD27, ICOS, NKG2D, DAP10, DAP12, 2B4 (CD244), BTLA, CD30, GITR, CD226, CD79A, and HVEM; and / or (ii) one or more intracellular signaling domains selected from the group consisting of CD3ζ and FC epsilon RI.

8. The primary cell, the iPSC or the derivative cell according to claim 7, wherein the one or more costimulatory signaling domains are selected from the group consisting of CD28, 41BB, and OX40 (CD134).

9. The primary cell, the iPSC or the derivative cell according to claim 7 or 8, wherein the one or more costimulatory domains comprise (i) 41BB, (ii) CD28, (iii) 41BB and CD28, (iv) 41BB and OX40, or (v) 41BB, CD28, and OX40.

10. The primary cell, the iPSC or the derivative cell according to any one of claims 1- 9, wherein: (i) the one or more CARs consists of a dual-targeting CAR comprising a first antigen-binding domain targeting a first tumor antigen and a second antigen binding domain targeting a second tumor antigen; or (ii) the one or more CARs comprises a plurality of CARs including a first CAR and a second CAR, wherein the first CAR comprises the first antigen binding domain targeting a first tumor antigen, and wherein the second CAR comprises a second antigen binding domain targeting a second tumor antigen.

11. The primary cell, the iPSC or the derivative cell according to any one of claims 1- 10, wherein the first tumor antigen is selected from Table 1.

12. The primary cell, the iPSC or the derivative cell according to claim 10 or 11, wherein the second tumor antigen is selected from Table 1.Docket No.: 066461.11126 / 25WO1 13. The primary cell, the iPSC or the derivative cell according to any one of claims 1- 12, wherein (i) each of the one or more CARs comprise amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to a sequence independently selected from the group consisting of SEQ ID NOs: 234-240, and 248-255, or (ii) each of the one or more CARs are encoded by a polynucleotide having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to a sequence independently selected from the group consisting of SEQ ID NOs: 241-247, and 256-263.

14. The primary cell, the iPSC or the derivative cell according to any one of claims 1- 13, wherein the first antigen binding domain comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one of SEQ ID NOs: 2, 4, and 158-193.

15. The primary cell, the iPSC or the derivative cell according to any one of claims 1- 13 wherein the first antigen binding domain is encoded by a polynucleotide having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one of SEQ ID NOs: 194-229.

16. The primary cell, the iPSC or the derivative cell according to any one of claims 10-15, wherein the second antigen binding domain comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one of SEQ ID NOs: 2, 4, and 158-193.

17. The primary cell, the iPSC or the derivative cell according to any one of claims 10-15, wherein the second antigen binding domain is encoded by a polynucleotide having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one of SEQ ID NOs: 194-229.

18. The primary cell, the iPSC or the derivative cell according to any one of claims 6- 17, wherein in the CAR, at least one of:Docket No.: 066461.11126 / 25WO1 (i) a signal peptide comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 1 or 264; (ii) the at least one extracellular domain comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one or more of SEQ ID NOs: 2, 4, and 158- 193; (iii) the GPCR scaffold comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one or more of SEQ ID NOs: 134, 234-240, and 266-281; (iv) the intracellular domain comprises a co-stimulatory domain (i) comprising amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one or more of SEQ ID NOs: 6, 8-20, and 156, or (ii) encoded by a polynucleotide having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one or more of SEQ ID NOs: 152- 155.

19. The primary cell, the iPSC or the derivative cell according to claim 18, wherein in the CAR, at least one of: (i) the signal peptide comprises amino acids having the sequence of SEQ ID NO: 1 or 264; (ii) the at least one extracellular domain comprises amino acids having the sequence of one of SEQ ID NOs: 2, 4, and 158-193; (iii) the GPCR scaffold comprises amino acids having the sequence of SEQ ID NO: 134, 234-240 and 266-281; (iv) the intracellular domain comprises a co-stimulatory domain (i) comprising amino acids having the sequence of one or more of SEQ ID NOs: 6, 8-20, and 156, or (ii) encoded by a polynucleotide having the sequence of one or more of SEQ ID NOs: 152-155.Docket No.: 066461.11126 / 25WO1 20. The primary cell, the iPSC or the derivative cell thereof according to any one of claims 1-19, wherein the cytokine comprises an interleukin 15 (IL-15) or a membrane bound IL-15, where all or a functional portion of the IL-15 protein is fused to all or a portion of a transmembrane protein that anchors the expressed IL- 15 as a cell membrane-bound polypeptide (mbIL15) 21. The primary cell, the iPSC or the derivative cell thereof according to claim 20, wherein the IL-15 comprises an inactivated cell surface receptor that comprises a monoclonal antibody-specific epitope and an IL-15, and wherein the inactivated cell surface receptor and the IL-15 are operably linked by an autoprotease peptide.

22. The primary cell, the iPSC or the derivative cell thereof according to claim 20, wherein the IL-15 comprises (i) a fusion polypeptide comprising an IL-15 and an IL-15 receptor alpha (IL-15Rα), or (ii) a polypeptide having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:

282.

23. The primary cell, the iPSC or the derivative cell thereof according to any one of claims 20-22, wherein the IL-15 comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:

72.

24. The primary cell, the iPSC or the derivative cell according to any one of claims 1- 23, comprising the deletion or reduced expression of one or more of B2M, TAP 1, TAP 2, Tapasin, RFXANK, CIITA, RFX5 and RFXAP genes.

25. The primary cell, the iPSC or the derivative cell according to any one of claims 1- 24, further comprising an exogenous polynucleotide encoding a human leukocyte antigen E (HLA-E) and / or human leukocyte antigen G (HLA-G).Docket No.: 066461.11126 / 25WO1 26. The primary cell, the iPSC or the derivative cell thereof according to any one of claims 1-25, wherein the CD16 is a CD16 variant protein.

27. The primary cell, the iPSC or the derivative cell thereof according to claim 26, wherein the CD16 variant protein is a high affinity CD16 variant.

28. The primary cell, the iPSC or the derivative cell thereof according to claim 26 or 27, wherein the CD16 variant protein is a non-cleavable CD16 variant.

29. The primary cell, the iPSC or the derivative cell thereof according to any one of claims 26-28, wherein the CD16 variant protein comprises wild-type CD16 comprising amino acids having the sequence of SEQ ID NO: 86 and one or more amino acid substitutions selected from the group consisting of F158V, F176V, S197P, D205A, S219A, T220A.

30. The primary cell, the iPSC or the derivative cell thereof according to any one of claims 26-29, wherein the CD16 variant protein comprises amino acids having at least 90% sequence identity to any one of SEQ ID NOs: 85 and 86.

31. The primary cell, the iPSC or the derivative cell thereof according to any one of claims 1-30 comprising an exogenous polynucleotide encoding the CD16 protein and the NKG2D protein, wherein the CD16 protein and the NKG2D protein are operably linked by an autoprotease peptide.

32. The primary cell, the iPSC or the derivative cell thereof according to claim 31, wherein the NKG2D protein is a wildtype NKG2D protein.

33. The primary cell, the iPSC or the derivative cell thereof according to claim 31, wherein the NKG2D protein comprises amino acids having at least 90% sequence identity to SEQ ID NO: 89.Docket No.: 066461.11126 / 25WO1 34. The primary cell, the iPSC or the derivative cell thereof according to any one of claims 31-33, wherein the autoprotease peptide is selected from the group consisting of a porcine tesehovirus-12A (P2A) peptide, a foot-and-mouth disease virus 2A (F2A) peptide, an Equine Rhinitis A Virus (ERAV) 2A (E2A) peptide, a Thosea asigna virus 2A (T2A) peptide, a cytoplasmic polyhedrosis virus 2A (BmCPV2A) peptide, and a Flacherie Virus 2A (BmIFV2A) peptide.

35. The primary cell, the iPSC or the derivative cell thereof according to claim 34, wherein the autoprotease peptide is a P2A peptide comprising amino acids having at least 90% sequence identity to SEQ ID NO:

91.

36. The primary cell, the iPSC or the derivative cell thereof according to any one of claims 31-35, wherein the exogenous polynucleotide encoding the CD16 protein and the NKG2D protein comprises a nucleic acid having at least 90% sequence identity to SEQ ID NO:

91.

37. The primary cell, the iPSC or the derivative cell thereof according to any one of claims 1-36 comprising (i) an exogenous polynucleotide encoding a membrane- bound interleukin 12 (IL-12) comprising a first polypeptide comprising an IL-12 alpha subunit p35 or a polypeptide at least 90% similar thereto, a second polypeptide comprising an IL-12 beta subunit p40 or a polypeptide at least 90% similar thereto, and a transmembrane domain fused to the terminus of the first and / or second IL-12 subunit polypeptide.

38. The primary cell, the iPSC or the derivative cell according to any one of claims 1- 37, wherein the exogenous polynucleotide(s) are integrated into the chromosome of the cell at one or more loci selected from the group consisting of AAVS1, B2M, CBL-B, CCR5, CD33, CD38, CD70, CIITA, CIS, CISH, CLYBL, Collagen, CTLA4 , GAPDH, HTRP, LAG3, NKG2A, NKG2D, NLRC5, PD1, RFX5, RFXANK, RFXAP, ROSA26, RUNX1, SOCS2, TAP2, Tapasin, TAPBP, TAPI, TCR a or b constant region, TIGIT, TIM3, and TRAC genes, provided at least one of the exogenous polynucleotides is integrated at a locus of a geneDocket No.: 066461.11126 / 25WO1 selected from the group consisting of AAVS1, B2M, TAP 1, TAP 2, Tapasin, RFXANK, CIITA, RFX5 and RFXAP genes to thereby result in a deletion or reduced expression of the gene.

39. The primary cell, the iPSC or the derivative cell according to claim 38, wherein the exogenous polynucleotide(s) are integrated into the chromosome of the cell at one or more loci selected from the group consisting of AAVS1, B2M, CIITA, CD33, and CLYBL.

40. The primary cell, the iPSC or the derivative cell according to any one of claims 1- 39 having a deletion or reduced expression of one or both of the B2M and the CIITA genes.

41. The primary cell, the iPSC or the derivative cell thereof according to claim 40, having a deletion or reduced expression of both of the B2M and the CIITA genes.

42. The primary cell of any one of claims 1-41, wherein the primary cell is a T-cell or an NK cell.

43. The primary cell of claim 42, wherein the primary cell is a T-cell.

44. The iPSC of any one of claims 1-41, wherein the iPSC is reprogrammed from whole peripheral blood mononuclear cells (PBMCs).

45. The iPSC of any one of claim 1-41, which is derived from a re-programmed T- cell.

46. The iPSC of any one of claims 1-41, 44, and 45, the derivative cell of any one of claims 1-41, or the primary cell of claim 42 or 43, further comprising an exogenous polynucleotide encoding a safety switch.Docket No.: 066461.11126 / 25WO1 47. The primary cell, the iPSC or the derivative cell thereof of claim 46, wherein the safety switch comprises an inactivated cell surface receptor that comprises a monoclonal antibody-specific epitope.

48. The primary cell, the iPSC or the derivative cell according to claim 47, wherein the monoclonal antibody specific epitope is selected from a group of epitopes specifically recognized by ibritumomab, tiuxetan, muromonab-CD3, tositumomab, abciximab, basiliximab, brentuximab vedotin, cetuximab, infliximab, rituximab, alemtuzumab, bevacizumab, certolizumab pegol, daclizumab, eculizumab, efalizumab, gemtuzumab, natalizumab, omalizumab, palivizumab, polatuzumab vedotin, ranibizumab, tocilizumab, trastuzumab, vedolizumab, adalimumab, belimumab, canakinumab, denosumab, golimumab, ipilimumab, tremelimumab, ofatumumab, panitumumab, and ustekinumab.

49. The primary cell, the iPSC or the derivative cell according to claim 47, wherein the inactivated cell surface receptor is a truncated epithelial growth factor (tEGFR) variant.

50. The primary cell, the iPSC or the derivative cell according to claim 49, wherein the tEGFR variant consists of amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:

71.

51. The primary cell, the iPSC or the derivative cell thereof according to any one of claims 1-50, wherein the safety switch comprises (i) an intracellular domain comprising a herpes simplex virus thymidine kinase (HSV-TK) or (ii) an inducible Caspase 9 (iCasp9).

52. The primary cell, the iPSC or the derivative cell thereof according to any one of claims 1-51 comprising the exogeneous polynucleotide encoding the PSMA cell tracer, wherein the PSMA cell tracer comprises an extracellular domain comprising a PSMA extracellular domain or fragment thereof.Docket No.: 066461.11126 / 25WO1 53. The primary cell, the iPSC or the derivative cell thereof according to claim 52, comprising a combined artificial cell death / reporter system polypeptide comprising an intracellular domain comprising a herpes simplex virus thymidine kinase (HSV-TK) and a linker, a transmembrane region, and an extracellular domain comprising the PSMA extracellular domain or fragment thereof.

54. The primary cell, the iPSC or the derivative cell thereof according to any one of claims 51-53, wherein (i) the HSV-TK comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 283 or 284, or (ii) the iCasp9 comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 285 or 286.

55. The primary cell, the iPSC or the derivative cell thereof according to claim 53, wherein the combined artificial cell death / reporter system polypeptide comprises the HSV-TK fused to a truncated variant PSMA polypeptide via the linker.

56. The primary cell, the iPSC or the derivative cell thereof according to claim 55, wherein the truncated variant PSMA polypeptide comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:

287.

57. The primary cell, the iPSC or the derivative cell thereof according to any one of claims 53-56, wherein the linker comprises an autoprotease peptide selected from the group consisting of a P2A peptide, a T2A peptide, an E2A peptide, and a F2A peptide.

58. The primary cell, the iPSC or the derivative cell thereof according to any one of claims 53-57, wherein the artificial cell death / reporter system polypeptide comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NOs: 288.Docket No.: 066461.11126 / 25WO1 59. The primary cell, the iPSC or the derivative cell thereof according to claim 58, wherein the artificial cell death / reporter system polypeptide comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 289-291.

60. The primary cell, the iPSC or the derivative cell thereof according to any one of claims 53-59, wherein the artificial cell death / reporter system polypeptide is encoded by nucleic acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 292-294.

61. The primary cell, the iPSC or the derivative cell according to any one of claims 1- 60, wherein the HLA-E comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: , or the HLA-G comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:

69.

62. The primary cell, the iPSC or the derivative cell according to any one of claims 2- 61, wherein: (i) the exogenous polynucleotide encodes one or more CARs comprising amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to at least one sequence selected from the group consisting of SEQ ID NOs: 2, 4, 158-193, 234-240, and 248-255; (ii) the exogenous polynucleotide encoding a human leukocyte antigen E (HLA-E) and / or human leukocyte antigen G (HLA-G) comprises polynucleotides having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one or more of SEQ ID NOs: 67 and 70;Docket No.: 066461.11126 / 25WO1 (iii) the exogenous polynucleotide encoding an NK cell receptor immunoglobulin gamma Fc region receptor III (FcyRIII, cluster of differentiation 16 (CD16)) and / or an NKG2D protein comprises polynucleotides having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one or more of SEQ ID NOs: 84, 88, and 90; (iv) the exogeneous polynucleotide encoding a cytokine comprises polynucleotides having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 295; (v) the exogenous polynucleotide encoding a safety switch comprises a polynucleotide having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one or more of SEQ ID NO: 292-294; and / or (vi) the exogeneous polynucleotide encoding a PSMA cell tracer comprises a polynucleotide sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:

287.

63. The primary cell, the iPSC or the derivative cell thereof according to any one of claims 1-62, wherein: (i) the exogenous polynucleotide encodes one or more CARs comprising amino acids having at least one sequence selected from the group consisting of SEQ ID NOs: 2, 4, 158-193, 234-240, and 248-255; (ii) the exogenous polynucleotide encoding a human leukocyte antigen E (HLA-E) and / or human leukocyte antigen G (HLA-G) comprises polynucleotides having the sequence SEQ ID NO: 67 and 70; (iii) the exogenous polynucleotide encoding an FcyRIII receptor and / or an NKG2D protein comprises polynucleotides having the sequence of SEQ ID NO: 84, 88, and 90;Docket No.: 066461.11126 / 25WO1 (iv) the exogeneous polynucleotide encoding the cytokine comprises polynucleotides having the sequence of SEQ ID NO: 295; and / or (v) the exogenous polynucleotide encoding the safety switch comprises polynucleotides having the sequence of one of SEQ ID NOs: 292-294; and / or (vi) the exogenous polynucleotide encoding the PSMA cell tracer comprises polynucleotides having the sequence of one of SEQ ID NOs:

287.

64. The primary cell, the iPSC or the derivative cell thereof according to claim 62 or 63, wherein each of the exogenous polynucleotides are integrated into the chromosome of the cell at a locus independently selected from the group consisting of AAVS1, B2M, CBL-B, CCR5, CD33, CD38, CD70, CIITA, CIS, CISH, CLYBL, Collagen, CTLA4 , GAPDH, HTRP, LAG3, NKG2A, NKG2D, NLRC5, PD1, RFX5, RFXANK, RFXAP, ROSA26, RUNX1, SOCS2, TAP2, Tapasin, TAPBP, TAPI, TCR a or b constant region, TIGIT, TIM3, and TRAC.

65. The primary cell, the iPSC or the derivative cell thereof according to claim 64, wherein: (i) the exogenous polynucleotide encoding the one or more CARs is integrated at a locus of the AAVS1 gene, the CD33 gene, or the CLYBL gene; (ii) the exogenous polynucleotide encoding a HLA-E)and / or HLA-G is integrated at a locus of the B2M gene; (iii) the exogenous polynucleotide encoding an FcyRIII receptor and / or an NKG2D protein is integrated at a locus of the CD70 gene; (iv) the exogeneous polynucleotide encoding the cytokine is integrated at the locus of the NKG2A gene; (v) The primary cell, the iPSC or the derivative cell thereof comprises a deletion or reduced expression of the CIITA gene; and, optionallyDocket No.: 066461.11126 / 25WO1 (vi) the exogenous polynucleotide encoding the safety switch or the PSMA is integrated at the locus of the CIITA gene.

66. The derivative cell of any one of claims 1-65, wherein the derivative cell is a natural killer (NK) cell or a T cell.

67. The derivative cell according to claim 66, wherein the derivative cell is a natural killer (NK) cell.

68. The derivative cell according to claim 66, wherein the derivative cell is a T cell.

69. The derivative cell according to claim 68, wherein the T cell is a gamma delta T cell.

70. The derivative cell according to claim 68, wherein the T cell is a gamma delta Vγ9 / Vδ1 T cell.

71. A composition comprising the iPSC according to any one of claims 1-65 or the derivative cell according to any one of claims 1-41 and 46-70.

72. The composition according to claim 71, further comprising or being used in combination with, one or more therapeutic agents selected from the group consisting of a small-molecule therapeutic agent, a biologic, a peptide, a cytokine, a checkpoint inhibitor, a mitogen, a growth factor, a small RNA, a dsRNA (double stranded RNA), a siRNA, an oligonucleotide, mononuclear blood cells, a vector comprising one or more polynucleotides of interest, an antibody, a chemotherapeutic agent, a radioactive moiety, and an immunomodulatory drug (IMiD).

73. A chimeric antigen receptor (CAR) polypeptide comprising amino acids encoding: (i) a G protein-coupled receptor (GPCR) scaffold or fragment thereof; andDocket No.: 066461.11126 / 25WO1 (ii) a first antigen binding domain targeting a tumor antigen.

74. The CAR according to claim 73, wherein the GPCR scaffold or fragment thereof is selected from the group consisting of rhodopsin-like receptors (Class A), secretin family receptors (Class B), or Metabotropic receptors (Class C).

75. The CAR according to claim 74 comprising the rhodopsin-like receptors (Class A), wherein the GPCR scaffold is a chemokine receptor selected from the group consisting of CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CCR10, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CXCR6, CX3CR1, XCR1, ACKR1, ACKR2, ACKR3, ACKR4, and a fragment of any receptor thereof.

76. The CAR according to claim 75, wherein the GPCR scaffold is CXCR3 or a fragment thereof.

77. The CAR according to claim 76, wherein (i) the CXCR3 comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 134, or (ii) the CXCR3 is encoded by a polynucleotide having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one or more of SEQ ID NOs: 135-151.

78. The CAR according to any one of claims 73-77, wherein the GPCR scaffold comprises: (i) at least one an extracellular domain comprising the first antigen- binding domain, wherein the extracellular domain comprises an extracellular loop of the GPCR scaffold; (ii) at least one transmembrane domain; and (iii) at least one intracellular domain.

79. The CAR according to claim 78, wherein the at least one intracellular domain comprises:Docket No.: 066461.11126 / 25WO1 (i) one or more costimulatory signaling domains selected from the group consisting of CD28, 41BB, IL2Rb, CD40, OX40 (CD134), CD80, CD86, CD27, ICOS, NKG2D, DAP10, DAP12, 2B4 (CD244), BTLA, CD30, GITR, CD226, CD79A, and HVEM; and / or (ii) one or more intracellular signaling domains selected from the group consisting of CD3ζ and FC epsilon RI.

80. The CAR according to claim 79, wherein the one or more costimulatory signaling domains are selected from the group consisting of CD28, 41BB, and OX40 (CD134).

81. The CAR according to claim 79 or 80, wherein the one or more costimulatory domains comprise (i) 41BB, (ii) CD28, (iii) 41BB and CD28, (iv) 41BB and OX40, or (v) 41BB, CD28, and OX40.

82. The CAR according to any one of claims 73-81, wherein the CAR consists of a dual-targeting CAR comprising the first antigen-binding domain and a second antigen binding domain targeting a second tumor antigen.

83. The CAR according to any one of claims 73-82, wherein the first tumor antigen is selected from Table 1.

84. The CAR according to claim 82 or 83, wherein the second tumor antigen is selected from Table 1.

85. The CAR according to any one of claims 73-84, wherein (i) the CAR comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one of SEQ ID NOs: 234-240, and 248-255, or (ii) the CAR is encoded by a polynucleotide having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one of SEQ ID NOs: 241-247, and 256-263.Docket No.: 066461.11126 / 25WO1 86. The CAR according to any one of claims 73-85, wherein the first antigen binding domain comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one of SEQ ID NOs: 2, 4, and 158-193.

87. The CAR according to any one of claims 73-85 wherein the first antigen binding domain is encoded by a polynucleotide having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one of SEQ ID NOs: 194-229.

88. The CAR according to any one of claims 82-87, wherein the second antigen binding domain comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one of SEQ ID NOs: 2, 4, and 158-193.

89. The CAR according to any one of claims 82-87, wherein the second antigen binding domain is encoded by a polynucleotide having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one of SEQ ID NOs: 194-229.

90. The CAR according to any one of claims 78-89, wherein in the CAR, at least one of: (i) the signal peptide comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 1 or 264; (ii) the at least one extracellular domain comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one or more of SEQ ID NOs: 2, 4, and 158- 193;Docket No.: 066461.11126 / 25WO1 (iii) the GPCR scaffold comprises amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 134, and 266-281; (iv) the intracellular domain comprises a co-stimulatory domain (i) comprising amino acids having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one or more of SEQ ID NOs: 6, 8-20, and 156, or (ii) encoded by a polynucleotide having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one or more of SEQ ID NOs: 152- 155.

91. The CAR according to claim 90, wherein in the CAR, at least one of: (i) the signal peptide comprises amino acids having the sequence of SEQ ID NO: 1 or 264; (ii) the extracellular domain comprises amino acids having the sequence of one of SEQ ID NOs: 2, 4, and 158-193; (iii) the GPCR scaffold comprises amino acids having the sequence of SEQ ID NO: 134, and 266-281; (iv) the intracellular domain comprises a co-stimulatory domain (i) comprising amino acids having the sequence of one or more of SEQ ID NOs: 6, 8-20, and 156, or (ii) encoded by a polynucleotide having the sequence of one or more of SEQ ID NOs: 152-155.

92. A method of treating cancer in a subject in need thereof, comprising administering the derivative cell according to any one of claims 1-41 and 46-70, or the composition according to claim 71 or 72 to a subject in need thereof.

93. The method of treatment according to claim 92, wherein the cancer is selected from the group consisting of leukemias, such as AML, CML, acute lymphocytic leukemia (ALL), B-cell acute lymphocytic leukemia (B-ALL), and chronic lymphocytic leukemia (CLL), lymphomas, such as Hodgkin lymphoma, non-Docket No.: 066461.11126 / 25WO1 Hodgkin lymphoma, multiple myeloma, and follicular lymphoma, and solid cancers such as sarcomas, skin cancer, melanoma, bladder cancer, brain cancer, breast cancer, uterus cancer, ovary cancer, prostate cancer, lung cancer, colorectal cancer, cervical cancer, liver cancer, head and neck cancer, esophageal cancer, pancreatic cancer, renal cancer, adrenal cancer, stomach cancer, testicular cancer, cancer of the gall bladder and biliary tracts, thyroid cancer, thymus cancer, cancer of bone, and cerebral cancer, as well as cancer of unknown primary (CUP).

94. The method of treatment according to claim 93, wherein the cancer is a B-cell malignancy, acute lymphocytic leukemia (ALL), B-cell acute lymphocytic leukemia (B-ALL), chronic lymphocytic leukemia (CLL), or non-Hodgkin lymphoma, follicular lymphoma.

95. The method of treatment according to any one of claims 92-94, wherein the subject has minimal residual disease (MRD) after an initial cancer treatment.

96. The method of treatment according to any one of claims 92-94, wherein the subject has no minimal residual disease (MRD) after one or more cancer treatments or repeated dosing.

97. A method of manufacturing the derivative cell according to any one of claims 1- 41 and 46-70 comprising differentiating the iPSC according to any one of claims 1-41 and 44-65 under conditions for cell differentiation to thereby obtain the derivative cell.

98. The method according to claim 97, wherein the iPSC is obtained by genomic engineering an unmodified iPSC, wherein the genomic engineering comprises targeted editing.

99. The method according to claim 97, wherein the targeted editing comprises deletion, insertion, or in / del carried out by CRISPR, ZFN, TALEN, homingDocket No.: 066461.11126 / 25WO1 nuclease, homology recombination, or any other functional variation of these methods.

100. A method of differentiating an induced pluripotent stem cell (iPSC) into an NK cell, comprising subjecting the iPSCs to a differentiation protocol including culturing the cells in a medium containing a recombinant human IL-12 for the final 24 hours of culturing under the differentiation protocol.

101. The method according to claim 100, wherein the recombinant IL-12 comprises IL12p70.

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