Targeting payloads

Abstract

The present disclosure provides technologies that achieve local delivery and / or retention of payloads or other active agents through use of y8 TCR sequences as targeting moieties. The present disclosure provides certain improvements in the deli very of compositions, e.g., that may be or comprise or otherwise deliver one or more payloads (which payloads may, for example, be or comprise or deliver an active agent) to, and / or retention of such payloads in, target tissue(s) of interest.

Description

TARGETING PAYLOADSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to United States Provisional Application Serial No. 63 / 618,244 filed January 05, 2024, the entirety of which is incorporated herein by reference.BACKGROUND

[0002] One challenge with many therapeutic agents is the inability to sufficiently localize them to a site (e.g., tissue) of interest.SUMMARY

[0003] The present disclosure, among other things, provides an insight that y5 T cells, unlike many other T cell types, often localize to and / or are retained within tissues. Some yS T cells preferentially localize to one or a small number of tissues; some may be considered tissue specific. For example, some y5 T cells preferentially localize to and / or are retained in a particular tissue in that they are several fold more enriched in such tissue compared to in other tissues.

[0004] The present disclosure further provides an insight that targeting moieties within T cell receptors (TCRs) of y8 TCRs may contribute to and / or achieve observed targeting and / or retention of y5 T cells within tissues.

[0005] The present disclosure further provides an insight that y5 TCR targeting moieties may be coupled to other payloads (i.e., to generate an engineered composition that comprises and / or delivers a pay load of interest), and can contribute to and / or achieve targeting and / or retention of such payload (or composition) in a relevant tissue of interest.

[0006] Among other things, the present disclosure provides engineered compositions comprising (i) a y8 TCR targeting moiety associated with, for example by covalent linkage with (ii) a payload; in some embodiments such paylod, for example, is or comprises or delivers an active agent.

[0007] In some embodiments, the present disclosure provides methods of localizing a payload to a tissue of interest and / or of retaining a payload in a tissue of interest, such method comprising a step of associating the payload with a y5 TCR targeting moiety.

[0008] Still further, the present disclosure provides methods of treating a disease, disorder or condition associated with a particular tissue in a subject, for example comprising a step of administering to the subject a composition comprising a payload (e.g., that may be or comprise or otherwise deliver an active agent) linked to a y8 TCR targeting moiety that localizes to the tissue; such provided technologies, in some embodiments, causes the active agent to be delivered and / or localized to a site at which it is useful in treating the disease, disorder or condition.

[0009] The present disclosure provides certain improvements in the delivery of compositions, e.g., that may be or comprise or otherwise deliver one or more payloads (which payloads may, for example, be or comprise or deliver an active agent) to, and / or retention of such payloads in, target tissue(s) of interest, which improvements in some embodiments comprise associating the composition (e.g., the pay load) to a y5 TCR targeting moiety so that the composition preferentially localizes to and / or is preferentially retained in a target tissue of interest relative to an otherwise comparable composition not so coupled.

[0010] In some embodiments, the present disclosure provides, in a therapeutic composition, the improvement that comprises associating the composition to a y5 TCR targeting moiety so that the composition preferentially localizes to and / or is preferentially retained in a target tissue of interest relative to an otherwise comparable composition not so coupled.

[0011] In some embodiments, an engineered composition comprises a y8 TCR targeting moiety; associated with, for example by covalent linkage with, a payload.

[0012] In some embodiments, the present disclosure provides a method of localizing a payload to a tissue of interest, the method comprising a step of associating the payload with a y8 TCR targeting moiety.

[0013] In some embodiments, the present disclosure provides a method of treating a disease, disorder or condition associated with a particular tissue in a subject, the method comprising a step of administering to the subject a composition comprising a payload linked to a y8 TCR targeting moiety that localizes to the tissue, wherein the payload is or comprises an active agent, or otherwise produces and / or causes the active agent to be delivered and localized to a site at which it is useful in treating the disease, disorder or condition.BRIEF DESCRIPTION OF THE DRAWING

[0014] Figure 1 depicts a provided strategy for using y8 TCR sequences as targeting moieties as described herein, referred to as TAGHOME™.

[0015] Figure 2 illustrates a therapeutic challenge addressed by provided technologies.

[0016] Figure 3 documents an insight provided by the present disclosure, namely that y8 TCRs direct y8 T Cells to distinct tissues; the present disclosure appreciates that such targeting can be harnessed as described herein to achieve delivery of payloads including therapeutics.

[0017] Figure 4 depicts a solution provided by an insight of the present disclosure, namely that y5 TCR ectodomain sequences can be utilized as targeting moieties to delivery pay loads preferentially (and / or specifically) to particular tissues (e.g., to diseased tissues) as described herein.

[0018] Figure 5 depicts a provided platform technology that identifies and / or characterizes particularly useful yd TCR sequences (e.g., y8 TCR ectodomain sequences) useful as targeting moieties in accordance with the present disclosure.

[0019] Figure 6 illustrates a particular application of provided technologies, which is to achieve gut-specific delivery of anti-inflammatory cytokine payloads, for example to treat inflammatory bowel disease (IBD).

[0020] Figure 7 provides a rationale for a particular embodiment of the present disclosure, in which y8 TCR sequences (e.g., y8 TCR ectodomain sequences) that target the gut are linked with an IL-10 payload moiety; the provided product is useful, for example, in the treatment of IBD.

[0021] Figure 8 depicts exemplary ex vivo and in vivo embodiments of provided technologies that utilize y8 TCR sequences (e. ., y8 TCR ectodomain sequences) to achieve targeted delivery of IL- 10 to the gut. In a particular depicted strategy, T cells engineered to express a TAGHOME™ y8 TCR sequence and / or an IL-10 payload as described herein are infused into a subject to achieve delivery of IL- 10 to the gut. In another depicted strategy, mRNA encoding a TAGHOME™-IL-10 payload expression construct is introduced (e.g., in vivo) into cells (e.g., y8 T-Cells), so that the targeted pay load is expressed (and / or that the cells expressing the payload themselves target the gut, e.g., via an anti-y4 TCR as depicted).

[0022] Figure 9 illustrates various conditions addressable by provided technologies, based on targeted tissues.

[0023] Figure 10 presents certain exemplary delta and gamma chain TCR sequences;CERTAIN DEFINITIONS

[0024] In general, terminology used herein is in accordance with its understood meaning in the art, unless clearly indicated otherwise. Explicit definitions of certain terms are provided below; meanings of these and other terms in particular instances throughout this specification will be clear to those skilled in the art from context.

[0025] In order that the present invention may be more readily understood, certain terms are first defined below. Additional definitions for the following terms and other terms are set forth throughout the specification.

[0026] Administration: As used herein, the term “administration” typically refers to the administration (e.g., of a composition or treatment) to a subject or system (e.g., that is or comprises one or more cells, tissues, organisms, etc), for example to achieve delivery of an agent that is, is included in, or is otherwise delivered or generated by, such composition ortreatment. In some embodiments, an administered composition on treatment includes the agent to be delivered. In some embodiments, an administered composition or treatment may include a precursor (e.g., a prodrug of the agent, a cell that produces the agent, a nucleic acid that encodes the agent, etc) of the agent, such that the agent is released or generated upon administration of the composition or treatment.

[0027] Affinity. As is known in the art, “affinity" is a measure of the tightness with which two or more binding partners associate with one another. Those skilled in the art are aware of a variety of assays that can be used to assess affinity, and will furthermore be aware of appropriate controls for such assays. In some embodiments, affinity is assessed in a quantitative assay. In some embodiments, affinity is assessed over a plurality of concentrations (e.g., of one binding partner at a time). In some embodiments, affinity is assessed in the presence of one or more potential competitor entities (e.g., that might be present in a relevant - e.g., physiological - setting). In some embodiments, affinity is assessed relative to a reference (e.g., that has a known affinity above a particular threshold [a “positive control" reference] or that has a known affinity below a particular threshold [ a “negative control" reference”]. In some embodiments, affinity may be assessed relative to a contemporaneous reference; in some embodiments, affinity may be assessed relative to a historical reference. Typically, when affinity is assessed relative to a reference, it is assessed under comparable conditions.

[0028] Agent: As used herein, the term “agent”, may refer to a physical entity or phenomenon. In some embodiments, an agent may be characterized by a particular feature and / or effect. In some embodiments, as will be clear from context, the term agent may be used to refer to an entity (e.g., for example, a lipid, metal, nucleic acid, polypeptide, polysaccharide, small molecule, etc, or complex, combination, mixture or system [e.g., cell, tissue, organism] thereof) and / or a phenomenon (e.g., heat, electric current or field, magnetic force or field, etc, or combination thereof). In some embodiments, the term “agent” may refer to a compound, molecule, or entity that comprises a polymer. In some embodiments, the term may refer to a compound or entity that comprises one or more polymeric moieties. In some embodiments, the term “agent” may refer to a compound, molecule, or entity that is substantially free of a particular polymer or polymeric moiety. Insome embodiments, the term may refer to a compound, molecule, or entity that lacks or is substantially free of any polymer or polymeric moiety. In some embodiments, an agent may be or comprise a system or device. In some embodiments, an agent may be or comprise information, e.g., a piece or collection of input or output data. In some embodiments, an agent may be or comprise a force such as an electric force, a gravitational force, a magnetic force, etc.

[0029] Agonist: Those skilled in the art will appreciate that the term “agonist” may be used to refer to an agent (e.g., an entity or condition) or event whose presence, level, degree, type, or form correlates with increased level or activity of another agent (i.e., the agonized agent or the target agent). In some embodiments, an agonist may be direct (in which case it exerts its influence directly upon its target); in some embodiments, an agonist may be indirect (in which case it exerts its influence by other than binding to its target; e.g., by interacting with a regulator of the target, so that level or activity of the target is altered).

[0030] Analog: As used herein, the term “analog” refers to a substance that shares one or more particular structural features, elements, components, or moieties with a reference substance. Typically, an “analog” shows significant structural similarity with the reference substance, for example sharing a core or consensus structure, but also differs in certain discrete way(s). In some embodiments, an analog is a substance that can be generated from the reference substance, e.g., by chemical manipulation of the reference substance. In some embodiments, an analog is a substance that can be generated through performance of a synthetic process substantially similar to (e.g., sharing a plurality of steps with) one that generates the reference substance. In some embodiments, an analog is or can be generated through performance of a synthetic process different from that used to generate the reference substance.

[0031] Antagonist: Those skilled in the art will appreciate that the term “antagonist”, as used herein, may be used to refer to an agent (e.g., an entity or condition) or event whose presence, level, degree, type, or form correlates with decreased level or activity of another agent (i.e., the inhibited agent, or target). In some embodiments, an antagonist may be direct (in which case it exerts its influence directly upon its target); insome embodiments, an antagonist may be indirect (in which case it exerts its influence by other than binding to its target; e.g., by interacting with a regulator of the target, so that level or activity of the target is altered).

[0032] Antibody agent: As used herein, the term “antibody agent” refers to an agent that specifically binds to a particular antigen. In some embodiments, the term encompasses a polypeptide or polypeptide complex that includes immunoglobulin structural elements sufficient to confer specific binding. For example, in some embodiments, an antibody agent is or comprises a polypeptide whose amino acid sequence includes one or more structural elements recognized by those skilled in the art as a complementarity determining region (CDR), appreciating that there is more than one system available in the art for defining such CDRs, such different systems typically variying in their precise definition of CDR boundaries. In some embodiments, an antibody agent is or comprises a polypeptide whose amino acid sequence includes at least one CDR (e.g., at least one heavy chain CDR and / or at least one light chain CDR) that is substantially identical to one found in a reference antibody. In some embodiments, an included CDR is substantially identical to a reference CDR in that it is either identical in sequence or contains between 1-5 amino acid substitutions as compared with the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that it shows at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that it shows at least 96%, 96%, 97%, 98%, 99%, or 100% sequence identity with the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that at least one amino acid within the included CDR is deleted, added, or substituted as compared with the reference CDR but the included CDR has an amino acid sequence that is otherwise identical with that of the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that 1- 5 amino acids within the included CDR are deleted, added, or substituted as compared with the reference CDR but the included CDR has an amino acid sequence that is otherwise identical to the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that at least one amino acid within the included CDR issubstituted as compared with the reference CDR but the included CDR has an amino acid sequence that is otherwise identical with that of the reference CDR. In some embodiments an included CDR is substantially identical to a reference CDR in that 1-5 amino acids within the included CDR are deleted, added, or substituted as compared with the reference CDR but the included CDR has an amino acid sequence that is otherwise identical to the reference CDR. In some embodiments, an antibody agent is or comprises a polypeptide whose amino acid sequence includes structural elements recognized by those skilled in the art as an immunoglobulin variable domain. In some embodiments, an antibody agent in or comprises a polypeptide whose amino acid sequence includes structural elements recognized by those skilled in the art to correspond to CDRsl, 2, and 3 of an antibody variable domain; in some such embodiments, an antibody agent in or comprises a polypeptide or set of polypeptides whose amino acid sequence(s) together include structural elements recognized by those skilled in the art to correspond to both heavy chain and light chain variable region CDRs, e.g., heavy chain CDRs 1, 2, and / or 3 and light chain CDRs 1, 2, and / or 3. In some embodiments, an antibody agent is a polypeptide protein having a binding domain which is homologous or largely homologous to an immunoglobulin-binding domain. In some embodiments, an antibody agent may be or comprise a polyclonal antibody preparation. In some embodiments, an antibody agent may be or comprise a monoclonal antibody preparation. In some embodiments, an antibody agent may include one or more constant region sequences that are characteristic of a particular organism, such as a camel, human, mouse, primate, rabbit, rat; in many embodiments, an antibody agent may include one or more constant region sequences that are characteristic of a human. In some embodiments, an antibody agent may include one or more sequence elements that would be recognized by one skilled in the art as a humanized sequence, a primatized sequence, a chimeric sequence, etc. In some embodiments, an antibody agent may be a canonical antibody (e.g., may comprise two heavy chains and two light chains). In some embodiments, an antibody agent may be in a format selected from, but not limited to, intact IgA, IgG, IgE or IgM antibodies; bi- or multi- specific antibodies (e.g., Zybodies®, etc); antibody fragments such as Fab fragments, Fab’ fragments, F(ab’)2 fragments, Fd’ fragments, Fd fragments, and isolated CDRs or sets thereof; single chain Fvs; polypeptide-Fc fusions; single domain antibodies(e.g., shark single domain antibodies such as IgNAR or fragments thereof); cameloid antibodies; masked antibodies (e.g., Probodies®); Small Modular ImmunoPharmaceuticals (“SMIPs™ ); single chain or Tandem diabodies (TandAb®); VHHs; Anticalins®; Nanobodies® minibodies; BiTE®s; ankyrin repeat proteins or DARPINs®; Avimers®; DARTs; TCR-like antibodies;, Adnectins®; Affilins®; Trans-bodies®; Affibodies®; TrimerX®; Micro Proteins; Fynomers®, Centyrins®; and KALBITOR®s. In some embodiments, an antibody may lack a covalent modification (e.g., attachment of a glycan) that it would have if produced naturally. In some embodiments, an antibody may contain a covalent modification (e.g., attachment of a glycan, a payload [e.g., a detectable moiety, a therapeutic moiety, a catalytic moiety, etc], or other pendant group [e.g., poly-ethylene glycol, etc.].

[0033] Associated: Two events or entities are “associated” with one another, as that term is used herein, if the presence, level, degree, type and / or form of one is correlated with that of the other. In some embodiments, two or more entities are physically “associated” with one another if they interact, directly or indirectly, so that they are and / or remain in physical proximity with one another. In some embodiments, two or more entities that are physically associated with one another are covalently linked to one another; in some embodiments, two or more entities that are physically associated with one another are not covalently linked to one another but are non-covalently associated, for example by means of hydrogen bonds, van der Waals interaction, hydrophobic interactions, magnetism, and combinations thereof.

[0034] Binding: It will be understood that the term “binding”, as used herein, typically refers to a non-covalent association between or among two or more entities. “Direct” binding involves physical contact between entities or moieties; indirect binding involves physical interaction by way of physical contact with one or more intermediate entities. Binding between two or more entities can typically be assessed in one or more of a variety of contexts - for example, in some embodiments where interacting entities or moieties are studied in isolation, and / or in some embodiments where interacting entities or moieties are studied in the context of more complex systems (e.g., while covalently or otherwise associated with a carrier entity and / or in a biological system or cell). Bindingbetween two entities may be considered “specific” if, under the conditions assessed, the relevant entities are more likely to associate with one another than with other available binding partners. Those skilled in the art will be aware of circumstances where a particular degree of preference for one potential binding partner over another is required for binding to be deemed sufficiently “specific” for a particular purpose or situation.

[0035] Characteristic portion : As used herein, the term “characteristic portion”, in the broadest sense, refers to a portion of a substance whose presence (or absence) correlates with presence (or absence) of a particular feature, attribute, or activity of the substance. In some embodiments, a characteristic portion of a substance is a portion that is found in the substance and in related substances that share the particular feature, attribute or activity, but not in those that do not share the particular feature, attribute or activity. In certain embodiments, a characteristic portion shares at least one functional characteristic with the intact substance.

[0036] Comparable: As used herein, the term “comparable” refers to two or more agents (e.g., entities or set(s) of conditions), situations, etc., that may not be identical to one another but that are sufficiently similar to permit comparison therebetween so that one skilled in the art will appreciate that conclusions may reasonably be drawn based on differences or similarities observed. In some embodiments, comparable sets of conditions, circumstances, individuals, or populations are characterized by a plurality of substantially identical features and one or a small number of varied features. Those of ordinary skill in the art will understand, in context, what degree of identity is required in any given circumstance for two or more such agents, entities, situations, sets of conditions, etc to be considered comparable. For example, those of ordinary skill in the art will appreciate that sets of circumstances, individuals, or populations are comparable to one another when characterized by a sufficient number and type of substantially identical features to warrant a reasonable conclusion that differences in results obtained or phenomena observed under or with different sets of circumstances, individuals, or populations are caused by or indicative of the variation in those features that are varied.

[0037] Corresponding to: As used herein, the term “corresponding to” refers to a relationship between two or more entities. For example, the term “corresponding to” may be used to designate the position / identity of a structural element in a compound or composition relative to another compound or composition (e.g., to an appropriate reference compound or composition). For example, in some embodiments, a monomeric residue in a polymer (e.g., an amino acid residue in a polypeptide or a nucleic acid residue in a polynucleotide) may be identified as “corresponding to” a residue in an appropriate reference polymer. For example, those of ordinary skill will appreciate that, for purposes of simplicity, residues in a polypeptide are often designated using a canonical numbering system based on a reference related polypeptide, so that an amino acid "corresponding to" a residue at position 190, for example, need not actually be the 190thamino acid in a particular amino acid chain but rather corresponds to the residue found at 190 in the reference polypeptide; those of ordinary skill in the art readily appreciate how to identify "corresponding" amino acids. For example, those skilled in the art will be aware of various sequence alignment strategies, including software programs such as, for example, BLAST, CS-BLAST, CUSASW++, DIAMOND, FASTA, GGSEARCH / GLSEARCH, Genoogle, HMMER, HHpred / HHsearch, IDF, Infernal, KLAST, USEARCH, parasail, PSLBLAST, PSI-Search, ScalaBLAST, Sequilab, SAM, SSEARCH, SWAPHI, SWAPHLLS, SWIMM, or SWIPE that can be utilized, for example, to identify “corresponding” residues in polypeptides and / or nucleic acids in accordance with the present disclosure. Those of skill in the art will also appreciate that, in some instances, the term “corresponding to” may be used to describe an event or entity that shares a relevant similarity with another event or entity (e.g., an appropriate reference event or entity). To give but one example, a gene or protein in one organism may be described as “corresponding to” a gene or protein from another organism in order to indicate, in some embodiments, that it plays an analogous role or performs an analogous function and / or that it shows a particular degree of sequence identity or homology, or shares a particular characteristic sequence element.

[0038] Designed: As used herein, the term “designed” refers to a feature of having been conceived, created, and / or generated through an act of the hand of man. For example, the term may be used to refer to an agent (i) whose structure is or was selected by the handof man; (ii) that is produced by a process requiring the hand of man; and / or (iii) that is distinct from natural substances and other known agents.

[0039] Detectable entity. The term “detectable entity” as used herein refers to an element, molecule, functional group, compound, fragment or moiety that is detectable. Examples of detectable entities include, but are not limited to: various ligands, radionuclides (e.g.,3H,14C,18F,19F,32P,35S,1351,12SI,123I,64CU,187Re,n iIn,90Y,99mTc,177Lu,89Zr etc.), fluorescent dyes (for specific exemplary fluorescent dyes, see below), chemiluminescent agents (such as, for example, acridinum esters, stabilized dioxetanes, and the like), bioluminescent agents, spectrally resolvable inorganic fluorescent semiconductors nanocrystals (i.e., quantum dots), metal nanoparticles (e.g., gold, silver, copper, platinum, etc.) nanoclusters, paramagnetic metal ions, enzymes (for specific examples of enzymes, see below), colorimetric labels (such as, for example, dyes, colloidal gold, and the like), biotin, dioxigenin, haptens, and polypeptides for which antisera or monoclonal antibodies are available.

[0040] Engineered: In general, the term “engineered” refers to the aspect of having been manipulated by the hand of man. For example, a polynucleotide is considered to be “engineered” when two or more sequences that are not linked together in that order in nature are manipulated by the hand of man to be directly linked to one another in the engineered polynucleotide and / or when a particular residue in a polynucleotide is non-naturally occurring and / or is caused through action of the hand of man to be linked with an entity or moiety with which it is not linked in nature. For example, in some embodiments described and / or utilized herein, an engineered polynucleotide comprises a regulatory sequence that is found in nature in operative association with a first coding sequence but not in operative association with a second coding sequence, is linked by the hand of man so that it is operatively associated with the second coding sequence. Comparably, a polypeptide may be considered to be “engineered” if encoded by or expressed from an engineered polynucleotide, and / or if produced other than natural expression in a cell. Analogously, a cell or organism is considered to be “engineered” if it has been subjected to a manipulation, so that its genetic, epigenetic, and / or phenotypic identity is altered relative to an appropriate reference cell such as otherwise identical cell that has not been so manipulated. In someembodiments, the manipulation is or comprises a genetic manipulation, so that its genetic information is altered (e.g., new genetic material not previously present has been introduced, for example by transformation, mating, somatic hybridization, transfection, transduction, or other mechanism, or previously present genetic material is altered or removed, for example by substitution or deletion mutation, or by mating protocols). In some embodiments, an engineered cell is one that has been manipulated so that it contains and / or expresses a particular agent of interest (e.g., a protein, a nucleic acid, and / or a particular form thereof) in an altered amount and / or according to altered timing relative to such an appropriate reference cell. As is common practice and is understood by those in the art, progeny of an engineered polynucleotide or cell are typically still referred to as “engineered’' even though the actual manipulation was performed on a prior entity.

[0041] Payload: In general, the term “payload”, as used herein, refers to an agent (e.g., an entity) that may be delivered or transported by association with another entity. In some embodiments, such association may be or include a covalent linkage; in some embodiments such association may be or include non-covalent interaction(s). In some embodiments, association may be direct; in some embodiments, association may be indirect. The term “payload” is not limited to a particular chemical identity or type; for example, in some embodiments, a payload may be or comprise, for example, an entity of any chemical class including, for example, a lipid, a metal, a nucleic acid, a polypeptide, a saccharide (e.g., a polysaccharide), small molecule, or a combination or complex thereof. In some embodiments, a payload may be or comprise a biological modifier, a detectable agent (e.g., a dye, a fluorophore, a radiolabel, etc.), a detecting agent, a nutrient, a therapeutic agent, etc., or a combination thereof. In some embodiments, a payload may be or comprise a cell or organism, or a fraction, extract, or component thereof. In some embodiments, a payload may be or comprise a natural product in that it is found in and / or is obtained from nature; alternatively or additionally, in some embodiments, the term may be used to refer to one or more entities that is man-made in that it is designed, engineered, and / or produced through action of the hand of man and / or is not found in nature. In some embodiments, an payload may be or comprise an agent in isolated or pure form; in some embodiments, such agent may be in crude form.

[0042] Reference: As used herein describes a standard or control relative to which a comparison is performed. For example, in some embodiments, an agent, animal, individual, population, sample, sequence or value of interest is compared with a reference or control agent, animal, individual, population, sample, sequence or value. In some embodiments, a reference or control is tested and / or determined substantially simultaneously with the testing or determination of interest. In some embodiments, a reference or control is a historical reference or control, optionally embodied in a tangible medium. Typically, as would be understood by those skilled in the art, a reference or control is determined or characterized under comparable conditions or circumstances to those under assessment. Those skilled in the art will appreciate when sufficient similarities are present to justify reliance on and / or comparison to a particular possible reference or control.

[0043] Small molecule: As used herein, the term “small molecule” means a low molecular weight organic and / or inorganic compound. In general, a “small molecule” is a molecule that is less than about 5 kilodaltons (kD) in size. In some embodiments, a small molecule is less than about 4 kD, 3 kD, about 2 kD, or about 1 kD. In some embodiments, the small molecule is less than about 800 daltons (D), about 600 D, about 500 D, about 400 D, about 300 D, about 200 D, or about 100 D. In some embodiments, a small molecule is less than about 2000 g / mol, less than about 1500 g / mol, less than about 1000 g / mol, less than about 800 g / mol, or less than about 500 g / mol. In some embodiments, a small molecule is not a polymer. In some embodiments, a small molecule does not include a polymeric moiety. In some embodiments, a small molecule is not and / or does not comprise a polypeptide (e.g., is not an oligopeptide). In some embodiments, a small molecule is not and / or does not comprise a polynucleotide (e.g., is not an oligonucleotide). In some embodiments, a small molecule is not and / or does not comprise a polysaccharide; for example, in some embodiments, a small molecule is not a glycoprotein, proteoglycan, glycolipid, etc.). In some embodiments, a small molecule is not a lipid. In some embodiments, a small molecule is biologically active. In some embodiments, a small molecule is detectable (e.g., comprises at least one detectable moiety). In some embodiments, a small molecule is a therapeutic agent. Those of ordinary skill in the art, reading the present disclosure, will appreciate that certain small molecule compoundsdescribed herein may be provided and / or utilized in a form such as, for example, a crystal form, a salt form, a protected form, a pro-drug form, an ester form, an isomeric form (e.g., as an optical and / or structural isomer), an isotopic form, etc. Those of skill in the art will appreciate that certain small molecule compounds have structures that can exist in one or more steroisomeric forms. In some embodiments, such a small molecule may be utilized in accordance with the present disclosure in the form of an individual enantiomer, diastereomer or geometric isomer, or may be in the form of a mixture of stereoisomers; in some embodiments, such a small molecule may be utilized in accordance with the present disclosure in a racemic mixture form. Those of skill in the art will appreciate that certain small molecule compounds have structures that can exist in one or more tautomeric forms. In some embodiments, such a small molecule may be utilized in accordance with the present disclosure in the form of an individual tautomer, or in a form that interconverts between tautomeric forms. Those of skill in the art will appreciate that certain small molecule compounds have structures that permit isotopic substitution (e.g.,2H or3H for H;,nC,13C or14C for 12C; ,13N or15N for 14N;17O or18O for 160;36C1 for XXC;18F for XXF; 1311 for XXXI; etc). In some embodiments, such a small molecule may be utilized in accordance with the present disclosure in one or more isotopically modified forms, or mixtures thereof. In some embodiments, as will be clear to a skilled person readinreference to a particular small molecule compound may relate to a specific form of that compound. In some embodiments, a particular small molecule compound may be provided and / or utilized in a salt form (e.g., in an acid-addition or base-addition salt form, depending on the compound); in some such embodiments, the salt form may be a pharmaceutically acceptable salt form. In some embodiments, where a small molecule compound is one that exists or is found in nature, that compound may be provided and / or utilized in accordance in the present disclosure in a form different from that in which it exists or is found in nature. Those of ordinary skill in the art will appreciate that, in some embodiments, a preparation of a particular small molecule compound that contains an absolute or relative amount of the compound, or of a particular form thereof, that is different from the absolute or relative (with respect to another component of the preparation including, for example, another form of the compound) amount of the compound or form that is present in a reference preparationof interest (e.g., in a primary sample from a source of interest such as a biological or environmental source) is distinct from the compound as it exists in the reference preparation or source. Thus, in some embodiments, for example, a preparation of a single stereoisomer of a small molecule compound may be considered to be a different form of the compound than a racemic mixture of the compound; a particular salt of a small molecule compound may be considered to be a different form from another salt form of the compound; a preparation that contains only a form of the compound that contains one conformational isomer ((Z) or (E)) of a double bond may be considered to be a different form of the compound from one that contains the other conformational isomer ((E) or (Z)) of the double bond; a preparation in which one or more atoms is a different isotope than is present in a reference preparation may be considered to be a different form; etc.

[0044] Therapeutic agent: As used herein, the phrase “therapeutic agent” refers to an agent (e.g., an entity or condition) that, when administered to a subject, has a therapeutic effect and / or elicits a desired biological and / or pharmacological effect. In some embodiments, a therapeutic agent is a substance that can be used to alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and / or reduce incidence of one or more symptoms or features of a disease, disorder, and / or condition. In some embodiments, a “therapeutic agent” is an agent that has been or is required to be approved by a government agency before it can be marketed for administration to humans. In some embodiments, a “therapeutic agent” is an agent for which a medical prescription is required for administration to humans.

[0045] Therapeutic regimen'. A “therapeutic regimen”, as that term is used herein, refers to a dosing regimen whose administration across a relevant population may be correlated with a desired or beneficial therapeutic outcome.

[0046] Therapeutically effective amount: As used herein, the term “therapeutically effective amount” means an amount of an agent (e.g., an entity or condition) that elicits a desired biological response when delivered e.g., by administration of a composition or therapy that comprises or otherwise causes the agent to be delivered) to a subject as part of a therapeutic regimen. In some embodiments, a therapeutically effective amount is an amountthat is sufficient, when delivered to a subject suffering from or susceptible to a disease, disorder, and / or condition, to treat, diagnose, prevent, and / or delay the onset of one or more features of the disease, disorder, and / or condition. As will be appreciated by those of ordinary skill in this art, the effective amount in a particular situation may vary, for example, depending on such factors as the desired biological endpoint, the substance to be delivered, the target cell or tissue, etc. For example, the effective amount of compound in a formulation to treat a disease, disorder, and / or condition is the amount that alleviates, ameliorates, relieves, inhibits, prevents, delays onset of, reduces severity of and / or reduces incidence of one or more symptoms or features of the disease, disorder, and / or condition when administered in accordance with an appropriate regimen. In some embodiments, a therapeutically effective amount is administered in a single dose; in some embodiments, multiple unit doses are required to deliver a therapeutically effective amount. Those of ordinary skill in the art will appreciate that the term "therapeutically effective amount" does not in fact require successful treatment be achieved in a particular individual. Rather, a therapeutically effective amount may be that amount that provides a particular desired pharmacological response in a significant number of subjects when administered to patients in need of such treatment. In some embodiments, reference to a therapeutically effective amount may be a reference to an amount as measured in one or more specific tissues (e.g., a tissue affected by the disease, disorder or condition) or fluids (e.g., blood, saliva, serum, sweat, tears, urine, etc.). Those of ordinary skill in the art will appreciate that, in some embodiments, a therapeutically effective amount of a particular agent or therapy may be formulated and / or administered in a single dose. In some embodiments, a therapeutically effective agent may be formulated and / or administered in a plurality of doses, for example, as part of a dosing regimen.

[0047] Therapeutically effective amount: As used herein, the term “therapeutically effective amount” means an amount of an agent (e.g., an entity or condition) that elicits a desired biological response when delivered e.g., by administration of a composition or therapy that comprises or otherwise causes the agent to be delivered) to a subject as part of a therapeutic regimen. In some embodiments, a therapeutically effective amount is an amount that is sufficient, when delivered to a subject suffering from or susceptible to a disease,disorder, and / or condition, to treat, diagnose, prevent, and / or delay the onset of one or more features of the disease, disorder, and / or condition. As will be appreciated by those of ordinary skill in this art, the effective amount in a particular situation may vary, for example, depending on such factors as the desired biological endpoint, the substance to be delivered, the target cell or tissue, etc. For example, the effective amount of compound in a formulation to treat a disease, disorder, and / or condition is the amount that alleviates, ameliorates, relieves, inhibits, prevents, delays onset of, reduces severity of and / or reduces incidence of one or more symptoms or features of the disease, disorder, and / or condition when administered in accordance with an appropriate regimen. In some embodiments, a therapeutically effective amount is administered in a single dose; in some embodiments, multiple unit doses are required to deliver a therapeutically effective amount. Those of ordinary skill in the art will appreciate that the term "therapeutically effective amount" does not in fact require successful treatment be achieved in a particular individual. Rather, a therapeutically effective amount may be that amount that provides a particular desired pharmacological response in a significant number of subjects when administered to patients in need of such treatment. In some embodiments, reference to a therapeutically effective amount may be a reference to an amount as measured in one or more specific tissues (e.g., a tissue affected by the disease, disorder or condition) or fluids (e.g., blood, saliva, serum, sweat, tears, urine, etc.). Those of ordinary skill in the art will appreciate that, in some embodiments, a therapeutically effective amount of a particular agent or therapy may be formulated and / or administered in a single dose. In some embodiments, a therapeutically effective agent may be formulated and / or administered in a plurality of doses, for example, as part of a dosing regimen.

[0048] Variant: As used herein in the context of molecules, e.g., nucleic acids, proteins, or small molecules, the term “variant” refers to a molecule that shows significant structural identity with a reference molecule but differs structurally from the reference molecule, e.g., in the presence or absence or in the level of one or more chemical moieties as compared to the reference entity. In some embodiments, a variant also differs functionally from its reference molecule. In general, whether a particular molecule is properly considered to be a “variant” of a reference molecule is based on its degree of structuralidentity with the reference molecule. As will be appreciated by those skilled in the art, any biological or chemical reference molecule has certain characteristic structural elements. A variant, by definition, is a distinct molecule that shares one or more such characteristic structural elements but differs in at least one aspect from the reference molecule. To give but a few examples, a polypeptide may have a characteristic sequence element comprised of a plurality of amino acids having designated positions relative to one another in linear or three-dimensional space and / or contributing to a particular structural motif and / or biological function; a nucleic acid may have a characteristic sequence element comprised of a plurality of nucleotide residues having designated positions relative to on another in linear or three- dimensional space. In some embodiments, a variant polypeptide or nucleic acid may differ from a reference polypeptide or nucleic acid as a result of one or more differences in amino acid or nucleotide sequence and / or one or more differences in chemical moieties (e.g., carbohydrates, lipids, phosphate groups) that are covalently components of the polypeptide or nucleic acid (e.g., that are attached to the polypeptide or nucleic acid backbone). In some embodiments, a variant polypeptide or nucleic acid shows an overall sequence identity with a reference polypeptide or nucleic acid that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, or 99%. In some embodiments, a variant polypeptide or nucleic acid does not share at least one characteristic sequence element with a reference polypeptide or nucleic acid. In some embodiments, a reference polypeptide or nucleic acid has one or more biological activities. In some embodiments, a variant polypeptide or nucleic acid shares one or more of the biological activities of the reference polypeptide or nucleic acid. In some embodiments, a variant polypeptide or nucleic acid lacks one or more of the biological activities of the reference polypeptide or nucleic acid. In some embodiments, a variant polypeptide or nucleic acid shows a reduced level of one or more biological activities as compared to the reference polypeptide or nucleic acid. In some embodiments, a polypeptide or nucleic acid of interest is considered to be a “variant” of a reference polypeptide or nucleic acid if it has an amino acid or nucleotide sequence that is identical to that of the reference but for a small number of sequence alterations at particular positions. Typically, fewer than about 20%, about 15%, about 10%, about 9%, about 8%, about 7%, about 6%, about 5%, about 4%, about 3%, or about 2% of the residues in avariant are substituted, inserted, or deleted, as compared to the reference. In some embodiments, a variant polypeptide or nucleic acid comprises about 10, about 9, about 8, about 7, about 6, about 5, about 4, about 3, about 2, or about 1 substituted residues as compared to a reference. Often, a variant polypeptide or nucleic acid comprises a very small number (e.g., fewer than about 5, about 4, about 3, about 2, or about 1) number of substituted, inserted, or deleted, functional residues (i.e., residues that participate in a particular biological activity) relative to the reference. In some embodiments, a variant polypeptide or nucleic acid comprises not more than about 5, about 4, about 3, about 2, or about 1 addition or deletion, and, in some embodiments, comprises no additions or deletions, as compared to the reference. In some embodiments, a variant polypeptide or nucleic acid comprises fewer than about 25, about 20, about 19, about 18, about 17, about 16, about 15, about 14, about 13, about 10, about 9, about 8, about 7, about 6, and commonly fewer than about 5, about 4, about 3, or about 2 additions or deletions as compared to the reference. In some embodiments, a reference polypeptide or nucleic acid is one found in nature. In some embodiments, a reference polypeptide or nucleic acid is a human polypeptide or nucleic acid.DETAILED DESCRIPTION OF CERTAIN EMBODIMENTSCompositions

[0049] The present disclosure teaches, among other things, that coupling y5 TCR targeting moieties to compositions that are, comprise, or deliver payloads (e.g., therapeutic payloads) of interest, can contribute to or achieve preferential- or specific-tissue localization and / or retention of such compositions. y8 TCR Targeting Moieties

[0050] Gamma delta (y 8) T cells, an unconventional subset of T cells, possess unique structural and functional characteristics that distinguish them from their alpha beta (aP) counterparts. Predominantly located in tissues, yo T cells are formed during the embryonic stage and migrate directly into tissues, where they often reside permanently.These cells undergo V(D)J recombination, generating a diverse range of T cell receptors (TCRs) composed of various gamma (y) and delta (8) chains, such as TRGV1, TRGV4, TRGV9, TRDV1, TRDV2, and TRDV3.

[0051] Gamma delta T cell receptors (yS TCRs) are crucial components of the immune system, uniquely characterized by their formation, tissue enrichment, structure, and antigen engagement.

[0052] Formation and Tissue Enrichment: y5 TCRs are generated through V(D)J recombination, a process that creates diverse receptor repertoires. These receptors are enriched in various tissues, with distinct distributions: V51 TCRs are predominantly found in epithelial tissues, while V52 TCRs are more common in the blood.

[0053] Structure: A y8 TCR is composed of a gamma and a delta chain, each with constant and variable regions. These chains form a heterodimer at the cell surface, essential for antigen recognition. The variable regions contribute to the diversity of antigen recognition, enabling y8 T cells to respond to a wide range of pathogens and abnormal cells.

[0054] Antigen Engagement: Upon antigen engagement, y8 TCRs initiate a cascade of signaling events, leading to T cell activation and proliferation. Unlike a[3 TCRs, y6 TCRs can recognize antigens without the need for presentation by Major Histocompatibility Complex (MHC) molecules. This includes recognizing cell surface proteins such as butyrophilins, non-peptidic antigens, CDld presented lipid antigens and / or other stress- induced ligands, and / or phosphoantigens, highlighting their role in innate and adaptive immunity.

[0055] Functional Diversity: The unique properties of yS TCRs facilitate diverse functions in immune responses, from direct cytotoxicity against infected or malignant cells to modulating inflammatory responses. The present disclosure provides an insight that these features of y5 TCRs can be harnessed to provide valuable therapeutic interventions, particularly in cancer and infectious diseases.

[0056] y8 TCRs are adept at recognizing tissue-specific antigens. Notably, deltal (V81) TCRs are enriched in tissues compared to their delta2 (V62) counterparts. Recent advancements have shed light on their ligand recognition in tissues, particularly the bindingof V51Vy4 (deltal gamma4) TCRs to heterodimers of butyrophilin-like 3 (BTNL3) and BTNL8, which are predominantly expressed in the gut.

[0057] y8 TCRs demonstrate a broader range of antigen recognition than do aP TCRs, which primarily recognize peptides presented by major histocompatibility complex (MHC) molecules. For example, y8 TCRs can recognize certain to phosphoantigens and butyrophilin-like molecules, underlining their role in immune surveillance and the distinction between homeostasis and stress conditions. The ligand recognition mechanisms, particularly for BTNL molecules, have been a focus of recent research, offering insights into the interplay between y5 TCRs and their ligands.

[0058] The present disclosure appreciates that diversity in y8 TCR ligand recognition is a testament to the versatility and importance of y8 T cells in the immune system, and also renders y5 TCRs particularly useful as targeting moieties to deliver payloads, specifically including therapeutic payloads, to sites of interest as described herein.

[0059] Those skilled in the art will be aware of various y5 TCR sequences (e. ., ectodomain sequences or, more specifically, ligand recognition sequences) that are useful for use as and / or inclusion in targeting moieties as described herein. Figure 10 presents certain exemplary delta chain and gamma chain TCR sequences. Those skilled in the art, reading the present disclosure, will appreciate that its teachings are not limited to these particular sequences.

[0060] Structural features of y8 TCRs have been reviewed (see, for example, Allison & Garboczi Mol. Immunol. 38:1051, 2002; Chien & Konigshofer Immunol. Rev 2015:46, 2007), and specific ligands have been identified for many (see, for example, Beseke & Prinz Cell. Mol. Immunol 17:914, 2020); skilled artisans, reading the present disclosure will appreciate, however, that precise knowledge of specific ligands is not necessarily required for practice of technologies provided by the present disclosure. For example, the present disclosure provides a platform for identifying and / or characterizing y8 TCR sequences for tissue-preferential and / or tissue- specific binding; such characterization may be sufficient to identify and / or confirm usefulness of particular y8 TCR sequences (e.g., of natural or synthetic such sequences, where synthetic sequences will typically be or comprise sequencesthat are variants of natural sequences, or of other previously defined reference sequence(s) (e.g., that may have been documented to have a particular tissue-binding preference or specificity, particularly when assessed in competition with an alternative binder to the same tissue.Payloads and Active Agents

[0061] One feature of provided technologies is that they can be applicable to a variety of payloads. For example, in some embodiments a payload is or comprises an active agent of interest (e. ., a therapeutic or diagnostic agent). In some embodiments, a payload produces an active agent of interest, or otherwise causes such active agent to be delivered to a subject to whom the payload, associated with a targeting moiety as described herein, is administered (or delivered).

[0062] In some embodiments, an active agent (which may, in some embodiments, be a payload) is or comprises a detectable entity or moiety.

[0063] In some embodiments, an active agent (which may, in some embodiments, be a payload) is or comprises a therapeutic entity or moiety.

[0064] In many embodiments, association between a targeting moiety and a payload is covalent. In some embodiments, association between a targeting moiety and a payload is direct; in some embodiments, association between a targeting moiety and a payload is indirect (e.g., via a linker). In many embodiments, a targeting moiety is covalently associated with a pay load via a linker.

[0065] In some embodiments, a single targeting moiety may be associated with a plurality of payloads; in some such embodiments, all such payloads are the same whereas in other such embodiments, the single targeting moiety is associated with a plurality of different payloads.

[0066] In some embodiments, a payload is or comprises a small molecule, which small molecule may, in many embodiments, be or comprise an active agent (or precursor thereof).

[0067] In some embodiments, a payload is or comprises a metal, which metal may, in many embodiments, be an active agent or component thereof.

[0068] In some embodiments, a payload is or comprises a radioisotope, which radioisotope may, in many embodiments, be an active agent or component or precursor thereof.

[0069] In some embodiments, a payload is or comprises a polypeptide (e.g., a detectable polypeptide such as a “tag” with respect to which a detection agent such as an antibody is known and available, or a therapeutic polypeptide, such as a cytokine or enzyme or hormone, or antibody agent, etc). Given that provided targeting moieties are polypeptide entities, in many embodiments in which the payload is or comprises a polypeptide, it is associated with a targeting moiety as a fusion polypeptide, optionally including one or more linker residues between the fused moieties. The present disclosure therefore provides such fusions, as well as nucleic acids that encode them, including RNA (e.g., mRNA) and / or DNA (e.g., single stranded or double- stranded DNA, for example a vector such as a plasmid or viral vector or other context that contains and / or expresses (e.g., when introduced into a host cell) such fusion polypeptide as is known in the art.

[0070] In some embodiments, a payload is or comprises a particle (e.g., a ceramic particle, a polymer particle, a lipid particle etc.) or vesicle (e.g., an optionally enveloped viral particle or virus-like-particle, a micelle, a liposome, an exosome, etc.). In some embodiments, one or more targeting moieties is covalently linked with a particle surface; in some such embodiments, such linkage may be via association with (e.g., as part of a fusion polypeptide) an entity wholly or partly on the particle surface. Where a particle is a biologic particle generated by cells (e.g., an exosome or other extracellular or intracellular vesicle), in some embodiments, a targeting moiety may be associated with such particle by expression of a nucleic acid encoding such targeting moiety (e.g., in the context of a fusion polypeptide in or on such particle) in the cell(s) from which the particle arises. In someembodiments, such targeting moiety will be associated with other elements that direct its expression particularly in or on the particle, for example on the particle surface. The present disclosure therefore provides cells that contain and / or express (e.g., are engineered to contain and / or express) such nucleic acids. In some embodiments, such nucleic acids (e.g., engineered such nucleic acids) may include one or more expression regulatory sequences that provides for a particular expression characteristic of the targeting moiety - e.g., constitutive expression or inducible expression and / or expression at a particular level and / or under certain conditions. Those skilled in the art are aware of a variety of such regulatory sequences, and also of useful and / or appropriate vectors in which they may be utilized.

[0071] In some embodiments, a payload is or comprises a cell; in some such embodiments, such cell comprises or otherwise delivers an agent of interest after administration of the cell (associated with a targeting moiety) to a subject. Analogous to the above discussion of biologic particles, in some embodiments, a targeting moiety may be associated with cell(s) by expression of a nucleic acid encoding such targeting moiety e.g., in the context of a fusion polypeptide in or on such particle) in the cell(s); relevant portions of that above discussion will be apparent to those skilled in the art and are incorporated herein by reference.

[0072] In some embodiments, a particle or cell payload includes or otherwise delivers an active agent of interest to a subject upon administration (or delivery) of such particle or cell payload associated with a targeting moiety as described herein to such subject.

[0073] Figure 8 presents depicts certain exemplary embodiments in which cells are engineered to be associated with provided targeting moieties and / or to express other payloads associated with targeting moieties, by expression of sequences encoding such targeting moieties within the cells. In some embodiments, relevant cells produce (e.g., are engineered to produce or naturally produce) an active agent of interest; as depicted in Figure 8, such active agent is a cytokine (IL- 10) useful in the treatment of certain inflammatory conditions such as, for example inflammatory bowel disease (IBD). Those skilled in the art will readily appreciate other indications with respect to which targeted delivery of IL- 10may be useful, and furthermore will appreciate other active agents and / or indications to which technologies as described herein, including as depicted in Figure 8, can usefully be applied.

[0074] Figure 8 specifically depicts certain ex vivo embodiments, in which cells are isolated from a subject and engineered to express a provided targeting moiety, an active agent of interest, or both, and then are returned to the subject. In such embodiments, a skilled person will appreciate that the targeting moiety is usefully selected and / or otherwise designed and / or engineered, to direct the cell(s) to a site expected to benefit from delivery of the relevant active agent. In some embodiments, relevant cells may already express a useful targeting moiety and / or active agent, such that engineering may be required only to achieve expression of one, and / or if desirable to increase or otherwise alter expression of that which is already (e.g., endogenously) expressed.

[0075] Figure 8 also specifically exemplifies certain in vivo embodiments, in which nucleic acids encoding a targeting moiety of interest (specifically, as depicted, in the context of a fusion with an active agent of interest) are administered to a subject for expression in cells within the subject that will secrete the fusion so that the targeting moiety can direct the active agent (which is also the payload in this embodiment) to a site at which it is expected to be beneficial. As noted above, the particular exemplified active agent in Figure 8 is IL- 10, and the specific exemplified indication is IBD; those skilled in the art will appreciate other indications and active agents to which these principles can be applied.

[0076] Furthermore, in Figure 8, at least the ex vivo cell therapy is depicted as utilizing T cells, and in particular gd T cells. Those skilled in the art, reading the present disclosure, will appreciate its applicability to a variety of cell types, and will understand relevant considerations for selection of appropriate cells in context of particular application(s) of provided technologies.Linkers

[0077] Those skilled in the art will appreciate that a variety of linkers are available for use when associating targeting moieties with payloads as described herein, andfurthermore will understand relevant considerations for selection of appropriate linker(s) in context of particular application(s) of provided technologies.

[0078] For example, those skilled in the art will be familiar with a range of linker technologies useful for chemical conjugation of a payload (e.g., a small molecule payload) with a targeting moiety as described herein. Furthermore, such skilled person will be aware of various features e.g., susceptibility to cleavage, for example, when exposed to a particular pH or enzyme or other feature of a site or milieu of interest) of available linkers that may be particularly useful in certain applications or embodiments of the present disclosure..

[0079] Alternatively or additionally, those skilled in the art will be aware, for example, of a variety of polypeptide linkers available in the art, and will appreciate that they may be particularly useful when both payload and targeting moiety are or comprise polypeptides, such that a single fusion polypeptide, including payload (or portion thereof), linker, and polypeptide can be expressed from a single nucleic acid. A skilled person will appreciate , of such available polypeptide linkers that may be particularly useful in certain contexts - including, for example, length, degree of flexibility e.g., rotational flexibility), susceptibility to cleavage (e.g., when exposed to a particular pH or enzyme or milieu).Characterization

[0080] Among other things, the present disclosure provides technologies that can achieve identification and / or characterization of targeting moieties (and / or conjugates containing them) of interest.

[0081] In some embodiments, characterization establishes a degree of binding (e.g., affinity and / or specificity) for a site (e.g., tissue or cell type) of interest.

[0082] To give but one example, Figure 5 depicts an exemplary platform for identification and / or characterization of useful targeting moieties in accordance with the present disclosure. In some embodiments, such targeting moieties have an amino acid sequence identical, or substantially identical, to that of sequences found in a reference TCR (see, e.g., Figure 10). In some embodiments, such targeting moieties have an amino acidsequence that differs from that found in such reference TCR, but remain sufficiently similar as to be appreciated by those skilled in the art as a variant of such reference TCR sequences.

[0083] Those skilled in the art, reading the present disclosure and aware of the literature relating to structure and binding characteristics of gd TCRs, and of relationships therebetween, including structure / function relationships, to design and / or select appropriate targeting moieties for use as described herein (e.g., for association with relevant pay loads).EQUIVALENTS

[0084] 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 invention described herein. The scope of the present invention is not intended to be limited to the above Description, but rather is as set forth in the following claims:

Claims

CLAIMSWe claim:

1. In a therapeutic composition, the improvement that comprises associating the composition to a y8 TCR targeting moiety so that the composition preferentially localizes to and / or is preferentially retained in a target tissue of interest relative to an otherwise comparable composition not so coupled.

2. An engineered composition comprising: a y8 TCR targeting moiety; associated with, for example by covalent linkage with, a payload.

3. A method of localizing a payload to a tissue of interest, the method comprising a step of: associating the payload with a y8 TCR targeting moiety.

4. A method of treating a disease, disorder or condition associated with a particular tissue in a subject, the method comprising a step of: administering to the subject a composition comprising: a payload linked to a y8 TCR targeting moiety that localizes to the tissue, wherein the payload is or comprises an active agent, or otherwise produces and / or causes the active agent to be delivered and localized to a site at which it is useful in treating the disease, disorder or condition.

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