Velocity receptor expressing immune cells and uses thereof

Abstract

Provided herein are chimeric polypeptides comprising (a) an extracellular domain comprising a cytokine-binding domain and a hinge domain; (b) an extracellular hinge domain; (c) a transmembrane domain; and (d) an intracellular signaling domain.

Description

[0001] VELOCITY RECEPTOR EXPRESSING IMMUNE CELLS AND USES THEREOF

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority to U.S. Provisional Patent Application No. 63 / 471,337, filed on June 6, 2023, and U.S. Provisional Patent Application No. 63 / 624,061, filed on January 23, 2024, which are incorporated herein by reference in their entireties.

[0004] SEQUENCE LISTING

[0005] This application contains a Sequence Listing that has been submitted electronically as an XML file named “44807-0460W01_ST26_SL.xml.” The XML file, created on June 4, 2024, is 255,634 bytes in size. The material in the XML file is hereby incorporated by reference in its entirety.

[0006] TECHNICAL FIELD

[0007] The present disclosure relates to the use of immune cells (e.g., T cells, B cells, NK cells) that have been genetically engineered to express velocity receptors (VRs) and the use of said VR- expressing immune cells to enhance their immunosurveillance toward cancers and pathogens. The disclosure also relates to methods and compositions for designing and producing such VR- expressing immune cells.

[0008] BACKGROUND

[0009] Restoring normal function to or enhancing the immune response is considered a branch of regenerative medicine. The method by which this is achieved clinically is termed immunotherapy. Adoptive cell transfer (ACT) of immune cells that have been genetically modified to express agents for tumor recognition, such as chimeric antigen receptors (CARs), is a type of targeted immunotherapy. Under the umbrella of what is termed cellular therapies, these modified immune cells have shown promise in the clinic for blood cancers (i.e., liquid tumors), specifically CAR-T cells, but fail to exhibit efficacy and safety in cancer patients with solid tumors (e.g., lung, ovarian cancer). One reason behind the discrepancy in response rates between targeting liquid and solid tumors is the requirement for complex immunosurveillance; targeting liquid tumors does not require it whereas targeting solid tumors does. Immunosurveillance in its most basic definition is the ability of immune cells to locate their target to carry out an immune response. Since the common route of administration of cellular therapies is intravenously, there is thus a natural ease for these therapies to locate liquid tumors as they will both occupy the bloodstream together. On the other hand, immunosurveillance to target solid tumors requires a far more complex procedure. The cellular therapies must extravasate from the bloodstream and migrate through the complex extracellular matrix and other tissue components to reach solid tumors prior to any recognition of the tumor that may occur. Once reaching a specific tumor periphery, the cellular therapy must also infiltrate and migrate throughout the tumor bulk to efficiently eradicate it. Hence, there exists an urgent need to enhance the motility of cellular therapies to lead to their clinical efficacy not only in cancer, but also in other diseases where the immune response proves critical. This is crucial, as the immune system is the body’s natural first line of response against sickness and disease and one of the main and major components of physiological healing and health maintenance. Hence, there exists a need in cellular therapies to exhibit heightened migration in modalities such as, but not limited to, cancer, autoimmune disease, wound healing, heart disease, and allergy.

[0010] SUMMARY

[0011] Provided herein are chimeric polypeptides including (a) an extracellular domain comprising a cytokine-binding domain; (b) an extracellular hinge domain; (c) a transmembrane domain; and (d) an intracellular signaling domain.

[0012] In some embodiments, the chimeric polypeptide comprises a single polypeptide. In some embodiments, the chimeric polypeptide comprises two polypeptides. In some embodiments, the chimeric polypeptide comprises more than two polypeptides. In some embodiments, the chimeric polypeptide further comprises a cleaving sequence, wherein the cleaving sequence couples the two or more polypeptides. In some embodiments, the cleaving sequence comprises a P2A, T2A, E2A, F2A, or IRES cleaving sequence.

[0013] In some embodiments, the cytokine-binding domain comprises an scFv. In some embodiments, the cytokine-binding domain comprises an lL5R-alpha, lL5R-beta, IFN-gammaRl, IFN-gammaR2, TNFR1, TNFR2 cytokine-binding domain, or any combination thereof. In some embodiments, the cytokine-binding domain specifically binds to IL5, IL8, IFN-gamma, TNF- alpha, or any combination thereof.

[0014] In some embodiments, the cytokine-binding domain specifically binds to IL5, wherein the chimeric polypeptide comprises SEQ ID NO: 93, SEQ ID NO: 16, SEQ ID NO: 84, SEQ ID NO: 17, SEQ ID NO: 85, SEQ ID NO: 73, SEQ ID NO: 78, SEQ ID NO: 12, SEQ ID NO: 15, SEQ ID NO: 21, and SEQ ID NO: 24. In some embodiments, the cytokine-binding domain specifically binds to IL8, wherein the chimeric polypeptide comprises SEQ ID NO: 93, SEQ ID NO: 69, SEQ ID NO: 84, SEQ ID NO: 70, SEQ ID NO: 85, SEQ ID NO: 73, SEQ ID NO: 78, SEQ ID NO: 12, SEQ ID NO: 15, SEQ ID NO: 21, and SEQ ID NO: 24. In some embodiments, the cytokinebinding domain specifically binds to IFN-gamma, wherein the chimeric polypeptide comprises SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, and SEQ ID NO: 33. In some embodiments, the cytokine-binding domain specifically binds to TFN-alpha, wherein the chimeric polypeptide comprises SEQ ID NO: 93, SEQ ID NO: 197, SEQ ID NO: 84, SEQ ID NO: 198, SEQ ID NO: 85, SEQ ID NO: 73, SEQ ID NO: 78, SEQ ID NO: 12, SEQ ID NO: 15, SEQ ID NO: 21, and SEQ ID NO: 24.

[0015] In some embodiments, the cytokine-binding domain is coupled to the intracellular signaling domain, wherein the intracellular signaling domain comprises an IL5R-alpha, IL5R-beta, IL8R-alpha, IL8R-beta, IFN-gammaRl, IFN-gammaR2, TNFR1, TNFR2, CD8-alpha, CD28 intracellular signaling domain. In some embodiments, the hinge domain comprises an IL5R-alpha, IL5R-beta, IFN-gammaRl, IFN-gammaR2, TNFR1, TNFR2, CD8-alpha, CD4, CD28, IgGl, IgG4 hinge domain, one or any combination or all of the four hinge domains of IL8R-alpha, or one or any combination or all of the four hinge domains of IL8R-beta. In some embodiments, the transmembrane domain comprises an IL5R-alpha, IL5R-beta, IFN-gammaRl, IFN-gammaR2, TNFR1, TNFR2, CD8-alpha, CD28 transmembrane domain, one or any combination or all of the seven transmembrane domains of IL8R-alpha, or one or any combination or all of the seven transmembrane domains of IL8R-beta. In some embodiments, the intracellular signaling domain comprises an IL5R-alpha, IL5R-beta, IFN-gammaRl, IFN-gammaR2, TNFR1, TNFR2 intracellular signaling domain, one or any combination or all of the four intracellular domains of IL8R-alpha, or one or any combination or all of the four intracellular domains of IL8R-beta.

[0016] Also provided herein are nucleic acids encoding any one of the chimeric polypeptides described herein.

[0017] Also provided herein are recombinant vectors comprising any one of the nucleic acids described herein. In some embodiments, the recombinant vector further comprises a promoter operationally linked to the nucleic acid. In some embodiments, the promoter is a constitutive promoter. In some embodiments, the promoter is an inducible promoter. In some embodiments, the promoter comprises an EFl -alpha, CMV, CAG, PgK, or SV40 promoter.

[0018] In some embodiments, the recombinant vector is a viral vector. In some embodiments, the viral vector is a lentiviral vector.

[0019] Also provided herein are immune cells comprising any one of the nucleic acids or any one of the recombinant vectors described herein.

[0020] Also provided herein are pharmaceutical compositions comprising any one of the chimeric polypeptides, any one of the nucleic acids, any one of the recombinant vectors, or any one of the immune cells described herein; and a pharmaceutically acceptable carrier.

[0021] Also provided herein are methods of treating a subject, the method comprising administering to the subject any one of the pharmaceutical compositions described herein, whereby treating a disease or a condition. In some embodiments, the disease or condition comprises cancer, autoimmune disease, wound healing, heart disease, or allergy. In some embodiments, the disease or condition comprises a solid cancer.

[0022] Also provided herein are methods of modulating immune cell migration, the method comprising delivering any one of the chimeric polypeptides, any one of the nucleic acids, any one of the recombinant vectors described herein into an immune cell. In some embodiments, the modulating comprises increasing immune cell migration. In some embodiments, the immune cell is a human immune cell. In some embodiments, the immune cell comprises a T cell, B cell, Natural Killer (NK) cell, dendritic cell (DC), macrophage, monocyte, eosinophil, basophil, neutrophil, mast cell, lymphoid progenitor cell, myeloid progenitor cell, hematopoietic stem cell, red blood cell, platelet, or megakaryocyte. In some embodiments, the immune cell comprises a T cell.

[0023] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although methods and materials similar or equivalent to those described herein can be used to practice the invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0024] The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims.

[0025] BRIEF DESCRIPTION OF DRAWINGS

[0026] FIG. 1 shows time-course 48-96 hour 2D cytotoxicity assay against luciferase-eGFP Raji lymphoma cells. T cells were either treated with lentivirus bearing a vector containing a CD 19- specific CAR and V5 spaced by a 2A sequence (FMC63 + V5), treated with a CD19-specific CAR alone (FMC63), or were not transduced (NTD). The CAR contained an FMC63 scFv.

[0027] FIG. 2 shows 48 hour 2D cytotoxicity assay against luciferase-mCherry 0VCAR3 ovarian cancer cells. T cells were either treated with a combination of V5 and a mesothelin-specific CAR with an M912 scFv (M912 + V5), were treated with the mesothelin CAR alone (M912), or were transduced with V5 alone (V5) which acted as a transduction control (see, e.g., WO 2015 / 188141 which is herein incorporated by reference in its entirety).

[0028] FIG. 3 shows 18-day 3D cytotoxicity assay against luciferase-mCherry OVCAR3 ovarian cancer cell organoids. T cells were either treated with a combination of V5 and a mesothelin-specific CAR with an M912 scFv (M912 + V5), were treated with the mesothelin CAR alone (M912), or were transduced with V5 alone (V5) which acted as a transduction control.

[0029] FIG. 4 shows 38-day in vivo NSG OVCAR3 ovarian cancer mouse model. Female NSG mice were injected subcutaneously in the right flank with 5 x 106luciferase-mCherry OVCAR3 cells in 100 pL 1 : 1 Matrigel in PBS. Tumors were allowed to grow until palpable (day 7). On day 8, mice were randomized after which 5 mice were then placed into each of the following groups except for the PBS group of which 4 mice were placed into the group. Mice were injected intravenously with 200 pL in the tail vein with either saline (PBS), or non-transduced T cells (NTD), M5 CAR-T cells (M5CAR alone), or T cells that were transduced with a combination therapy of V5 and M5 CAR (V5 + M5CAR). On day 27, mice received a second injection of 100 pL of the specified controls and treatment intraperitoneally in the left flank. Note that for both injections on days 8 and 27, 3 x 106NTD, M5CAR, or V5 + M5CAR cells were injected. For the day 8 injection, M5CAR alone and V5 + M5CAR cells were normalized to 45% positivity of the M5CAR, whereas they were normalized to 71% for the day 27 injection. Tumors were measured at least once weekly using digital calipers. Error is plotted as SEM, and two-tailed t-tests were performed between V5 + M5CAR and M5CAR alone (see, e.g., WO 2015 / 090230 which is herein incorporated by reference in its entirety).

[0030] FIG. 5 shows 38-day in vivo NSG OVCAR3 ovarian cancer mouse model. Female NSG mice were injected subcutaneously in the right flank with 5 x 106luciferase-mCherry OVCAR3 cells in 100 pL 1 : 1 Matrigel in PBS. Tumors were allowed to grow until palpable (day 7). On day 8, mice were randomized after which 5 mice were then placed into each of the following groups except for the PBS group of which 4 mice were placed into the group. Mice were injected intravenously with 200 pL in the tail vein with either saline (PBS), or non-transduced T cells (NTD), M5 CAR-T cells (M5CAR alone), or T cells that were transduced with a combination therapy of VR5a IL5 and M5 CAR (VR5a IL5 + M5CAR). On day 27, mice received a second injection of 100 pL of the specified controls and treatment intraperitoneally in the left flank. Note that for both injections on days 8 and 27, 3 x 106NTD, M5CAR alone, or VR5a 1L5 + M5CAR cells were injected. For the day 8 injection, M5CAR alone and VR5a IL5 + M5CAR cells were normalized to 45% positivity of the M5CAR, whereas they were normalized to 71% for the day 27 injection. Tumors were measured at least once weekly using digital calipers. Error is plotted as SEM, and two-tailed t-tests were performed between VR5a IL5 + M5CAR and M5CAR alone.

[0031] FIG. 6 shows 38-day in vivo NSG OVCAR3 ovarian cancer mouse model. Female NSG mice were injected subcutaneously in the right flank with 5 x 106luciferase-mCherry OVCAR3 cells in 100 pL 1 : 1 Matrigel in PBS. Tumors were allowed to grow until palpable (day 7). On day 8, mice were randomized after which 5 mice were then placed into each of the following groups except for the PBS group of which 4 mice were placed into the group. Mice were injected intravenously with 200 pL in the tail vein with either saline (PBS), or non-transduced T cells (NTD), M5 CAR-T cells (M5CAR alone), or T cells that were transduced with a combination therapy of VR5a IL8 and M5 CAR (VR5a IL8 + M5CAR). On day 27, mice received a second injection of 100 pL of the specified controls and treatment intraperitoneally in the left flank. Note that for both injections on days 8 and 27, 3 x 106NTD, M5CAR alone, or VR5a IL8 + M5CAR cells were injected. For the day 8 injection, M5CAR alone and VR5a IL8 + M5CAR cells were normalized to 45% positivity of the M5CAR, whereas they were normalized to 71% for the day 27 injection. Tumors were measured at least once weekly using digital calipers. Error is plotted as SEM, and two-tailed t-tests were performed between VR5a IL8 + M5CAR and M5CAR alone. FIG. 7 shows 38-day in vivo NSG OVCAR3 ovarian cancer mouse model. Female NSG mice were injected subcutaneously in the right flank with 5 x 106luciferase-mCherry OVCAR3 cells in 100 pL 1 : 1 Matrigel in PBS. Tumors were allowed to grow until palpable (day 7). On day 8, mice were randomized after which 5 mice were then placed into each of the following groups except for the PBS group of which 4 mice were placed into the group. Mice were injected intravenously with 200 pL in the tail vein with either saline (PBS), or non-transduced T cells (NTD), M5 CAR-T cells (M5CAR alone), or T cells that were transduced with a combination therapy of VySigand M5 CAR (Vysig+ M5CAR). On day 27, mice received a second injection of 100 pL of the specified controls and treatment intraperitoneally in the left flank. Note that for both injections on days 8 and 27, 3 x 106NTD, M5CAR alone, or Vysig+ M5CAR cells were injected. For the day 8 injection, M5CAR alone and Vysig+ M5CAR cells were normalized to 45% positivity of the M5CAR, whereas they were normalized to 71% for the day 27 injection. Tumors were measured at least once weekly using digital calipers. Error is plotted as SEM, and two-tailed t-tests were performed between Vysig+ M5CAR and M5CAR alone.

[0032] FIG. 8 shows 38-day in vivo NSG OVCAR3 ovarian cancer mouse model. Female NSG mice were injected subcutaneously in the right flank with 5 x 106luciferase-mCherry OVCAR3 cells in 100 pL 1 : 1 Matrigel in PBS. Tumors were allowed to grow until palpable (day 7). On day 8, mice were randomized after which 5 mice were then placed into each of the following groups except for the PBS group of which 4 mice were placed into the group. Mice were injected intravenously with 200 pL in the tail vein with either saline (PBS), or non-transduced T cells (NTD), M5 CAR-T cells (M5CAR alone), or T cells that were transduced with a combination therapy of VR5a TNFa and M5 CAR (VR5a TNFa + M5CAR). On day 27, mice received a second injection of 100 pL of the specified controls and treatment intraperitoneally in the left flank. Note that for both injections on days 8 and 27, 3 x 106NTD, M5CAR alone, or VR5a TNFa + M5CAR cells were injected. For the day 8 injection, M5CAR alone and VR5a TNFa + M5CAR cells were normalized to 45% positivity of the M5CAR, whereas they were normalized to 71% for the day 27 injection. Tumors were measured at least once weekly using digital calipers. Error is plotted as SEM, and two-tailed t-tests were performed between VR5a TNFa + M5CAR and M5CAR alone.

[0033] FIGs. 9A-9S show low and high-motility states of T cell migration in vitro regulated by specific endogenously expressed cytokines. (FIG. 9A) Current T cell therapies need to traverse a major physical barrier - the extracellular matrix (ECM) - in the tumor microenvironment (TME) of solid tumors for eventual cell-cell contact that they do not experience in the context of liquid tumors. (FIG. 9B) Primary human pan T cells were isolated from primary human PBMCs using a donor leukopak. Isolated T cells premixed with a CD2 / CD3 / CD28 activation solution and 100 lU / ml IL2 were encapsulated in 3D gels composed of the major ECM protein, type 1 collagen (rat tail type 1 collagen at 2 mg / ml), at increasing T cell densities. (FIG. 9C) After a 48-h incubation, the spontaneous motion of T cells at increasing densities (LD = 105 cells / ml, MD = 5x105 cells / ml, HD = 106 cells / ml) embedded in 3D collagen gels was monitored using time-lapsed phase-contrast microcopy for 1 h in 3-min intervals at 37°C and 5% CO2. Cells were manually tracked to generate trajectories for each cell. A custom software was used to convert trajectories into mean squared displacement (MSD) for each cell at a time lag of 9 min. Each dot represents an individual cell. LD = low density, MD = medium density, HD = high density. (FIG. 9D) A Matlab script was coded to extract the fractions of T cells moving more than their own size (>R2= 25 pm2, where R = 5 pm is the average T cell radius) from the MSDs shown in panel C. (FIG. 9E) Activated LD and HD T cells in 3D collagen gels were incubated for 48 h, after which medium from LD T cells was replaced with conditioned medium (CM) obtained from HD cells. (FIG. 9F) MSDs of LD, HD, and LD + HD CM T cells imaged and analyzed as in (FIG. 9C). (FIG. 9G) Motile fractions corresponding to panel F, computed as in panel D. (FIG. 9H) Molecules secreted by LD and HD T cell in 3D collagen gels incubated for 48 h. CM was extracted from the top of LD and HD T cell gels. Single-cell secretomics were run on CM to generate average intensity of a human adaptive immune cytokine panel using Isoplexis Isocode chips. (FIG. 91) Average intensity from (FIG. 9H) normalized per 105cells for the four cytokines identified from (FIG. 9H) with higher intensities for HD CM compared to LD CM. Data is plotted as mean with SEM (n = 2). One-way ANOVA with Dunn’s multiple comparison test was performed for statistical analysis. ****P < 0.0001. (FIG. 9 J) Activated LD T cells in 3D collagen gels were incubated for 48 h. 100 ng / ml IL5, TNFoc, IFNy, or IL8 were then exogenously added to LD T cell gels separately and incubated for 1 h at 37°C and 5% CO2. (FIG. 9K) MSDs of T cells in the presence of IL5, TNFa, IFNY,orIL8 and corresponding (FIG. 9L) motile fractions. (FIG. 9M) Activated HD T cells in 3D collagen gels were incubated for 48 h. 25 mg / ml anti-IL5, anti-TNFaRl, anti-ZFNyRl, or anti-IL8R were then exogenously added separately and incubated for 1 h at 37°C and 5% CO2. (FIG. 9N) MSDs of HD T cells incubated with different functional antibodies and (FIG. 90) corresponding motile fractions. (FIG. 9P) Activated HD T cells in 3D collagen gels incubated for 48 h. 2.5 mM anti- JAK2 (AG490), 60 mM anti-STATl (fludarabine), or 50 mM anti-STAT3 (S3I) were then exogenously added separately and incubated for 1 h at 37°C and 5% CO2. (FIG. 9Q) MSDs of HD T cell gels incubated with different inhibitors and corresponding (FIG. 9R) motile fractions. (FIG. 9S) T cells self-propel using velocity cytokines (VCs) IL5, TNFoc, IFNy, and IL8.

[0034] FIGs. 10A-10K show velocity receptors turn M5CAR T cells into self-propelling highly motile cells and improve their effector functions. (FIG. 10A) Exemplary design of velocity receptors (VRs) that bind VCs and signal through VC receptor signaling domains, and development of VR- M5CAR therapies. (FIG. 10B) M5CAR or VR-M5CAR T cells at low density (LD) were encapsulated in 3D collagen gels supplemented with 100 lU / ml IL2 and incubated for 48 h at 37°C and 5% CO2 and their migration patterns were analyzed as in FIGs. 9A-9S. (FIG. 10C) Control HD M5CAR T cells were also encapsulated in collagen gels and incubated for 48 h and compared to LD VR-M5CAR T cells. The spontaneous motion LD M5CAR, LD VR-M5CAR, and HD M5CAR T cells was monitored using time-lapsed phase-contrast microcopy for 1 h at 2-min intervals at 37°C and 5% CO2. Cells were manually tracked to generate trajectories for each cell. A custom software was used to convert trajectories into MSD for each cell at a time lag of 10 min. Each dot represents an individual cell. (FIG. 10D) Fractions of T cells moving more than their own size (>R2 = 25 pm2, where R = 5 pm is the average T cell radius), as assessed from the MSDs shown in panel C. (FIG. 10E) LD M5CAR cells expressing either VR5ocIL5, VR5ocTNFoc, or VR5aIL8, embedded in 3D collagen gels were incubated for 48 h. 25 pg / ml anti-IL5, anti-TNFoc, or anti-ZL8R were then exogenously added to VR5ocIL5-M5CAR, VR5aTNFoc-M5CAR, or VR5aIL8-M5CAR, respectively, and incubated for 1 h at 37°C and 5% CO2. (FIG. 10F) MSDs of control LD M5CAR T cells and the different VR-M5CAR T cells shown in (FIG. 10A). T cells were imaged for 30 min at 2-min intervals as in (FIG. 10C). Cell trajectories and MSD were measured and computed as in (FIG. 10C). (FIG. 10G) Motile fractions of T cells were obtained as in (FIG. 10D) for data shown in (FIG. 10F). For all experiments, MSD, median with the SEM are plotted (n=2 technical replicates per biological replicate, N=2 biological replicates for (FIG. 10C)). Individual dots represent individual cells; an average of at least 120 cells per technical replicate up to 269 cells per technical replicate were tracked. One-way ANOVA with Dunn’s multiple comparison test was used for statistical analysis (ns = not significant, *P < 0.05, **P < 0.01, ****p < 0.0001). For experiments measuring the motile fraction, mean with SEM is plotted (n=2 technical replicates per biological replicate, N=2 biological replicates) for (FIG. 10D), and mean is plotted (n=2 technical replicates) for (FIG. 10G). (FIG. 10H) Surface expression of the activation markers CD69 and CD25, as determined by flow cytometry using PE anti-CD69 and BV421 anti-CD25. Non-transduced (NTD) T cells were included as a negative control. Surface expression of CD69 was analyzed in FlowJo on M5CAR+-gated T cells as determined by GFP expression. (FIG. 101) Surface expression of the activation maker CD25 was determined by flow cytometry using BV421 anti-CD25. Flow cytometric analysis was performed as in (FIG. 10H). (FIG. 10J) In vitro T cell killing of mesothelin-expressing H226 and OVCAR3 cells at the indicated effector-to-target (E:T) ratios. Data plotted as mean ± SEM, n = 3. A two-tailed student’s t test was used for statistical analysis (*P < 0.05, **P < 0.01). (FIG. 10K) 112, TNFa, and IFNy secretion by non-transduced (NTD), M5CAR T cells, and VR-M5CAR T cells after overnight coincubation with mesothelin-expressing ASPC1, H226, and OVCAR3 cells. Cytokine release was measured using ELISA. Mean with SEM is shown (n = 3).

[0035] FIGs. 11A-11H show VRs greatly elevate M5CAR T cell tumor infiltration for controlling tumor growth in a pancreatic cancer model. (FIG. 11 A) 2e6 ASPC1 pancreatic cancer tumors pre-mixed in 1: 1 MatrigekPBS were subcutaneously (s.c.) engrafted into 8-12 week old NSG mice. Tumor volumes were measured twice a week using digital calipers. When tumors were palpable (100-250 mm3), mice were randomized and blindly treated 10 days later with a single intravenous (i.v.) dose of 3e6 NTD or M5CAR T cells. 25 days after tumor engraftment, tumors were harvested and sent for sectioning and IHC staining for human CD45 at the Johns Hopkins Oncology Tissue Services Core Facility. IHC-stained tissue slides were scanned, and images were extracted using ImageScope (middle panel). Tumor volumes were calculated as (L x W2) x 0.5 plotted as mm3(right panel). (FIG. 11B) ASPCl-bearing mice were treated with M5CAR alone or VR5oc TNFa-M5CAR T cells as in panel A. IHC images and tumor volumes were obtained as in panel A. (FIG. 11C) ASPCl-bearing mice were treated with M5CAR alone or VR5oc IL8-M5CAR T cells as in panel A. IHC images and tumor volumes were obtained as in panel A. (FIG. 11D) ASPC1- bearing mice were treated with M5CAR T cells alone or Vysig-M5CAR T cells as in panel A. IHC images and tumor volumes were obtained as in panel A. (FIG. HE) ASPCl-bearing mice were treated with M5CAR alone or VR5ot IL5-M5CAR T cells as in panel A. IHC images and tumor volumes were obtained as in panel A. (FIG. 11F) ASPCl-bearing mice were treated with M5CAR alone or V5-M5CAR T cells as in panel A. IHC images and tumor volumes were obtained as in panel A. (FIG. 11G) Human CD45+ cell numbers were obtained as described herein. Tissue area was computationally obtained. Tumor infiltration of human CD45+ cells per area was calculated and plotted, (n = 2 non-consecutive tissue slides were stained per treatment group). (FIG. 11H) Human CD45+ cells per tumor area were plotted against average tumor volume at the study endpoint. For figure panels A to F, IHC scale bars are 2 mm (left) and 100 pm (right). Tumor volumes are plotted as mean ± SEM (n = 5 mice per group, except for NTD for which n = 4).

[0036] FIGs. 12A-12H show VRs improve M5CAR T cell tumor infiltration which can slow tumor growth in a lung cancer model. (FIG. 12A) 5e6 H226 lung cancer tumors pre-mixed in 1 : 1 MatrigekPBS were engrafted s.c. into 8-12 week old NSG mice. Tumor volumes were measured twice a week using digital calipers. When tumors were palpable (100-250 mm3), mice were randomized and blindly treated (9 days later) with a single i.v. dose of 3e6 NTD or M5CAR T cells. 19 days after tumor engraftment, tumors were harvested and sent for sectioning and IHC staining for human CD45 at the Johns Hopkins Oncology Tissue Services Core Facility. IHC- stained tissue slides were scanned, and images were extracted using ImageScope (middle panel). Tumor volumes were calculated as (L x W2) x 0.5 plotted as mm3(right panel). (FIG. 12B) H226- bearing mice were treated with M5CAR alone or VR5a TNFa-M5CAR T cells as in panel A. IHC images and tumor volumes were obtained as in panel A. (FIG. 12C) Mice bearing H226 tumors were treated with M5CAR alone or Vysig-M5CAR T cells as in panel A. IHC images and tumor volumes were obtained as in panel A. (FIG. 12D) Mice bearing H226 tumors were treated with M5CAR alone or VR5oc IL8-M5CAR T cells as in panel A. IHC images and tumor volumes were obtained as in panel A. (FIG. 12E) H226-bearing mice were treated with M5CAR alone or V5-M5CAR T cells as in panel A. IHC images and tumor volumes were obtained as in panel A. (FIG. 12F) Mice bearing H226 tumors were treated with M5CAR alone or VR5a IL5-M5CAR T cells as in panel A. IHC images and tumor volumes were obtained as in panel A. (FIG. 12G) Human CD45+ cell numbers were obtained as described herein. Tissue area was computationally obtained. Tumor infdtration of human CD45+ cells per area was calculated and plotted, (n = 2 nonconsecutive tissue slides were stained per treatment group). (FIG. 12H) Human CD45+ cells per tumor area were plotted against average tumor volume at the study endpoint. For figure panels A to F, IHC panel scale bars are 2 mm (left) and 100 pm (right). Tumor volumes are plotted as mean ± SEM (n = 5 mice per group, except for NTD for which n = 4).

[0037] FIGs. 13A-13F show VRs-M5CAR T cells extend the overall survival of mice in an ovarian cancer model. (FIG. 13A) 5e6 OVCAR3 ovarian cancer tumor cells pre-mixed in 1: 1 MatrigekPBS were engrafted s.c. into 8-12 week old female NSG mice. Tumor volumes were measured twice a week using digital calipers. When tumors were palpable (100-250 mm3), mice were randomized and blindly treated (8 days later) with a single i.v. dose of 3e6 M5CAR alone or VR-M5CAR T cells. Mice were treated 19 days later with 3e6 M5CAR alone or VR-M5CAR T cells by intraperitoneal (i.p.) injection. Mice reached an endpoint when OVCAR3 tumors reached 750 mm3in size. (FIG. 13B) OVCAR3-bearing mice were treated with M5CAR alone or VR5oc TNFa-M5CAR T cells. Tumor volumes were obtained as described in (FIG. 13A). Mice that reached the endpoint tumor size as described in panel A were removed from the study. Plotted are Kaplan-Meier survival curves. (FIG. 13C) OVCAR3-bearing mice were treated with M5CAR alone or Vysig-M5CAR T cells. Kaplan-Meier survival curves were generated as described in panel B. (FIG. 13D) Mice bearing OVCAR3 tumors were treated with M5CAR alone or V5-M5CAR T cells. Kaplan-Meier survival curves were generated as described in panel B. (FIG. 13E) OVCAR3-bearing mice were treated with M5CAR alone or VR5a IL8-M5CAR T cells. Kaplan-Meier survival curves were generated as described in panel B. (FIG. 13F) Mice bearing OVCAR3 tumors were treated with M5CAR alone or VR5a IL5-M5CAR T cells Kaplan-Meier survival curves were generated as described in panel B. All plots show Kaplan-Meier survival curves (n = 5 mice per group).

[0038] FIG. 14A shows LD, MD, and HD T cells were encapsulated in 3D collagen gels and their motility was monitored as described in FIGs. 9B-9D. A custom Matlab software was used to randomly select 10 trajectories per condition. Plotted are x-y trajectories (pm). FIG. 14B shows median T cell MSD was calculated and plotted versus time lag from all cell trajectories. FIG. 14C shows T cells that moved more than their own size (>R2 = 25 pm2, where R ~ 5 pm is the average T cell radius) in an hour of tracking were extracted and the median MSD was calculated and plotted versus time lag (min). FIG. 14D shows a Matlab script was coded to extract the percentage of motile T cells MSDs at a time lag of 9 min from the MSDs in FIG. 14C. FIG. 14E shows LD, HD, and LD + HD T cells were encapsulated in 3D collagen gels and imaged as described in FIGs. 9E-9G. Ten randomly selected trajectories displayed per condition. FIG. 14F shows median T cell MSDs for the indicated conditions. FIG. 14G shows MSDs of motile T cells were extracted as described in FIG. 14C. FIG. 14H shows MSDs of motile T cells at a time lag of 9 min from data in FIG. 14G. FIG. 141 shows LD T cells in 3D collagen gels were treated with exogenous cytokines and their motility was monitored as described in FIGs. 10C-10E. Ten randomly selected trajectories (pm) per condition are shown. FIG. 14 J shows median T cell MSD was calculated and plotted for the indicated conditions. FIG. 14K shows MSDs of motile T cells were extracted as described in FIG. 14C. FIG. 14L shows MSDs of motile T cells at a time lag of 9 min, computed from MSDs, as described in FIG. 14C. FIG. 14M shows HD T cells in 3D collagen gels were treated with indicated antibodies and their motility was monitored as described in FIGs. 10F-10H. Ten randomly selected x-y trajectories (pm) per condition are shown. FIG. 14N shows HD T cells in 3D collagen gels were treated with an anti-IL8R antibody and their motility was monitored as described in FIGs. 10F-10H. Ten randomly selected x-y trajectories (pm) per condition are shown. FIG. 140 shows median T cell MSDs for the indicated conditions. FIG. 14P shows median T cell MSDs for HD control T cells or HD T cells treating with an anti-IL8R antibody. FIG. 14Q shows motile MSDs were extracted and plotted in Matlab as described in FIG. 14C. FIG. 14R shows Matlab was used to extract and plot motile T cell MSDs at a time lag of 9 min from the MSDs in FIG. 14Q. FIG. 14S shows MSDs of motile T cells were extracted as described in FIG. 14C. FIG. 14T shows MSDs of T cells at a time lag of 9 min shown in FIG. 14S. FIG. 14U shows HD T cells in 3D collagen gels were treated with indicated JAK / STAT inhibitors and their motility was monitored as described in FIGs. 10I-10K. Ten randomly selected trajectories per condition are shown. FIG. 14V shows median MSDs of T cells for the indicated conditions. FIG. 14W shows MSDs of motile T cells were extracted as described in FIG. 14C. FIG. 14X shows MSDs of T cells at a time lag of 9 min computed from the MSDs in FIG. 14W.

[0039] FIG. 15A shows an exemplary VR design. Calculated are the number of VR constructs obtained from the indicated combination of domains. FIG. 15B shows an exemplary VR-M5CAR T cell co-transduction scheme.

[0040] FIG. 16A shows CD4+ and CD8+ primary human T cells were isolated from Ficoll-extracted PBMCs from human donor leukopaks and mixed 1 : 1. T cells were then activated overnight with CD3 / CD28 Dynabeads and 100 lU / ml IL2. The following day, lentiviruses bearing plasmids with transgenes that encode for the specified VRs (see FIG. HA) were incubated on Retronectin-coated plates for 4 hrs at 37 °C and 5% CO2. Activated T cells were then added to lentivirus-bound Retronectin coated wells and incubated overnight. VR surface expression was detected the next day by flow cytometry after staining with anti-F(ab)2 antibodies for the VRs indicated, anti-IL5RA for V5, or anti-IFNGRl for Vgsig. Non-transduced (NTD) T cells were used as a negative control. FlowJo was used to analyze the data and plot histograms normalized to mode. FIG. 16B shows the day following VR transduction, T cells were transduced overnight with M5CAR-P2A-eGFP (see FIG. 11 A) on lentivirus-bound Retronectin-coated plates. The next day, M5CAR expression was measured by GFP detection using flow cytometry. Non-transduced (NTD) T cells were used as a negative control. Data was analyzed and histograms normalized to mode were plotted in FlowJo. FIG. 16C shows LD M5CAR, LD VR-M5CAR, and HD M5CAR T cells were encapsulated in 3D collagen gels and their motility was monitored as described in FIGs. 11B-11D. Ten randomly selected trajectories per condition were selected. FIG. 16D shows median MSD of T cells vs. time lag. FIG. 16E shows T cells that moved more than their own size (>R2 = 25 pm2, where R = 5 pm is the average T cell radius) in an hour of tracking were extracted in Matlab and median MSD was calculated and plotted versus time lag. FIG. 16F shows MSDs of motile T cells at a time lag of 10 min, as described in FIG. 16E. FIG. 16G shows LD M5CAR or LD VR5aIL5- M5CAR, VR5aTNFa-M5CAR, or VR5aZL8-M5CAR cells were suspended in 3D collagen gels, treated with antibody inhibitors, incubated, and their motility was monitored as described in FIGs. 11E-11G. Ten randomly selected trajectories (mm) per condition are shown. FIGs. 16H-16J show median T cell MSD was calculated and plotted using a custom Matlab software. FIGs. 16K-16M show motile MSDs were extracted and plotted in Matlab (as described in FIGs. 16H-16J). FIGs. 16N-16P show a custom Matlab script was used to extract and plot motile T cell MSDs at a time lag of 10 min from the MSDs in FIGs. 16K-16M.FIGs. 17A-17H show CD45+ IHC micrographs of tumors from ASPCl-bearing mice (corresponding to FIG. 12) treated with PBS (FIG. 17A), NTD T cells (FIG. 17B), M5CAR alone T cells (FIG. 17C), VR5aTNFa-M5CAR T cells (FIG. 17D), VR5aIL8-M5CAR T cells (FIG. 17E), Vgsig-M5CAR T cells (FIG. 17F), VR5aIL5- M5CAR T cells (FIG. 17G), or V5-M5CAR T cells (FIG. 17H). Scale bars in IHC micrographs are 2 mm.

[0041] FIGs. 18A-18C show tumor growth of individual mice corresponding to ASPCl-bearing mice (as in FIG. 12) (FIG. 18A), H226-bearing mice (as in FIG. 13) (FIG. 18B), and OVCAR3 -bearing mice (as in FIG. 14) (FIG. 18C) with indicated treatments. FIG. 18D shows mice were weighed at the end of the study. For FIGs. 18A-18C, n = 5 for all treatment groups, except for PBS and NTD for which n = 4 (FIG. 18A). n = 5 for all treatment groups, except for NTD for which n = 4 and PBS for which n = 3 (FIG. 18B). n = 5 for all treatment groups, except for PBS for which n = 4 (FIG. 18C). For FIG. 18D, body weights are plotted as mean ± SEM (n = 5 mice per group, except for PBS and NTD for which n = 4). Ordinary one-way ANOVA with Tukey’s multiple comparison test was used for statistical analysis (ns = not significant). FIG. 19A shows 2x106ASPC1 pancreatic cancer tumors pre-mixed in 1 : 1 MatrigekPBS were subcutaneously (s.c.) engrafted into 8-12 week old NSG mice. Tumor volumes were measured twice a week using digital calipers. When tumors were palpable (100 - 250 mm3), mice were randomized and blindly treated (8 days later) with a single intravenous (i.v.) dose of 3x106NTD or M5CAR T cells. Tumor volumes were calculated as (L x W2) x 0.5 plotted as mm3. FIG. 19B shows ASPCl-bearing mice were treated with M5CAR alone or VR5aTNFa-M5CAR T cells (as in FIG. 19A). FIG. 19C shows ASPCl-bearing mice were treated with M5CAR alone or VR5aIL8-M5CAR T cells (as in FIG. 19A). FIG. 19D shows ASPCl-bearing mice were treated with M5CAR alone or Vgsig-M5CAR T cells (as in FIG. 19A). FIG. 19E shows ASPCl-bearing mice were treated with M5CAR alone or VR5aIL5-M5CAR T cells (as in FIG. 19A). FIG. 19F shows ASPCl-bearing mice were treated with M5CAR alone or V5-M5CAR T cells (as in FIG. 19A). FIG. 19G shows mice were weighed at the end of the study. For all figure panels, tumor volumes (FIGs. 19A-19F) and body weights (FIG. 19G) are plotted as mean ± SEM (n = 5 mice per group, except for PBS for which n = 3). For FIGs. 19A-19F, two-tailed student’s t test was used for statistical analysis (ns = not significant, *P < 0.05, **P < 0.01). For FIG. 19G, ordinary one-way ANOVA with Tukey’s multiple comparison test was used for statistical analysis (ns = not significant).

[0042] FIG. 20A shows ASPC1 pancreatic tumors were subcutaneously engrafted into NSG mice and measured as described in FIG. 12. Mice bearing ASPC1 tumors were treated with M5CAR T cells as described in FIG. 12. The kidney, pancreas, and spleen were harvested from day-25 mice. Organs were sectioned and IHC-stained with anti-human CD45 to detect T cell therapies as in FIG. 12. FIG. 20B shows mice bearing ASPC1 tumors were treated with VR5a TNFa-M5CAR T cells as described in FIG. 12. The kidney, pancreas, and spleen were sectioned and IHC-stained with anti -human CD45 to detect T cell therapies as in FIG. 12. FIG. 20C shows ASPCl-bearing mice were treated with VR5a IL8-M5CAR T cells as described in FIG. 12. The kidney, pancreas, and spleen were sectioned and IHC-stained with anti-human CD45 for CAR detection. FIG. 20D shows ASPCl-bearing mice were treated with Vgsig-M5CAR T cells as described in FIG. 12. The kidney, pancreas, and spleen were sectioned and IHC-stained with anti-human CD45. FIG. 20E shows ASPCl-bearing mice were treated with VR5a IL5-M5CAR T cells as described in FIG. 12. The kidney, pancreas, and spleen were sectioned and IHC-stained with anti-human CD45 to detect human T cells as in FIG. 12. FIG. 20F shows ASPCl-bearing mice were treated with V5-M5CAR T cells as described in FIG. 12. The kidney, pancreas, and spleen were sectioned and IHC-stained with anti-human CD45 to detect T cell therapies as in FIG. 12. For FIGs. 20A-20F, scale bars in IHC micrographs are 500 pm. FIG. 20G shows human CD45+ cell numbers were obtained as described herein. Tissue area was computationally obtained. Organ and tumor infdtration of human CD45+ cells per area were calculated and plotted. For calculation of cells / cm2in organs, n = 6 for NTD or n = 8 for M5CAR alone and VR-M5CARs, with 2 non-consecutive tissue slides stained per mouse and 3-4 mice per group. For calculation of cells / cm2in tumors, n = 6 for NTD or n = 8 for VR-M5CARs, with 2 non-consecutive tissue slides stained per mouse and 3-4 mice per group; n = 7 for M5CAR alone, with 2 non-consecutive tissue slides stained for 3 mice and 1 tissue slide stained for 1 mouse. FIG. 20H shows mouse body weights were collected at the end of the study. Body weights are plotted as mean ± SEM (n = 5 mice per group, except for PBS and NTD for which n = 4). Ordinary one-way ANOVA with Tukey’s multiple comparison test was used for statistical analysis (ns = not significant).

[0043] FIGs. 21A-21H show CD45+ IHC micrographs of tumors from H226-bearing mice (corresponding to FIG. 13) treated with PBS (FIG. 21A), NTD T cells (FIG. 21B), M5CAR alone T cells (FIG. 21C), VR5aTNFa-M5CAR T cells (FIG. 21D), Vgsig-M5CAR T cells (FIG. 21E), VR5aIL8-M5CAR T cells (FIG. 21F), V5-M5CAR T cells (FIG. 21G), or VR5aIL5-M5CAR T cells (FIG. 21H). Scale bars in IHC micrographs are 2 mm.

[0044] FIG. 22A shows 5xl06H226 lung cancer tumors pre-mixed in 1 : 1 MatrigekPBS were engrafted s.c. into 8-12 week old NSG mice. Tumor volumes were measured twice a week using digital calipers. When tumors were palpable (100 - 250 mm3), mice were randomized and blindly treated (10 days later) with a single i.v. dose of 3xl06NTD or M5CAR T cells. Tumor volumes were calculated as (L x W2) x 0.5 plotted as mm3. FIG. 22B shows H226-bearing mice were treated with M5CAR alone or VR5aTNFa-M5CAR T cells (as in FIG. 22A). FIG. 22C shows mice bearing H226 tumors were treated with M5CAR alone or Vgsig-M5CAR T cells (as in FIG. 22A). FIG. 22D shows mice bearing H226 tumors were treated with M5CAR alone or VR5aIL8- M5CAR T cells (as in FIG. 22A). FIG. 22E shows H226-bearing mice were treated with M5CAR alone or V5-M5CAR T cells (as in FIG. 22A). FIG. 22F shows mice bearing H226 tumors were treated with M5CAR alone or VR5aIL5-M5CAR T cells (as in FIG. 22A). FIG. 22G shows mouse body weights were collected at the end of the study. For all figure panels, tumor volumes (FIGs. 22A-22F) and body weights (FIG. 22G) are plotted as mean ± SEM (n = 5 mice per group, except for PBS for which n = 2, NTD for which n = 3, and M5CAR alone and VR5aTNFa-M5CAR for which n = 4). For FIGs. 22A-22F, two-tailed student’s t test was used for statistical analysis (ns = not significant, *P < 0.05, **P < 0.01). For FIG. 22G, ordinary one-way ANOVA with Tukey’s multiple comparison test was used for statistical analysis (ns = not significant).

[0045] FIG. 23A shows H226 lung tumors were subcutaneously engrafted into NSG mice and measured as described in FIG. 13. Mice bearing H226 tumors were treated with M5CAR T cells as described in FIG. 13. The kidney, pancreas, and spleen were harvested from day- 19 mice. Organs were sectioned and IHC-stained with anti -human CD45 to detect T cell therapies as in FIG. 13. FIG. 23B shows mice bearing H226 tumors were treated with VR5a TNFa-M5CAR T cells as described in FIG. 13. The kidney, pancreas, and spleen were sectioned and IHC-stained with anti -human CD45 to detect T cell therapies as in FIG. 13. FIG. 23C shows H226-bearing mice were treated with Vgsig-M5CAR T cells as described in FIG. 13. The kidney, pancreas, and spleen were sectioned and IHC-stained with anti-human CD45 for CAR detection. FIG. 23D shows H226- bearing mice were treated with VR5a IL8-M5CAR T cells as described in FIG. 13. The kidney, pancreas, and spleen were sectioned and IHC-stained with anti-human CD45. FIG. 23E shows H226-bearing mice were treated with V5-M5CAR T cells as described in FIG. 13. The kidney, pancreas, and spleen were sectioned and IHC-stained with anti-human CD45 to detect T cell therapies as in FIG. 13. FIG. 23F shows mice bearing H226 tumors were treated with VR5a IL5- M5CAR T cells as described in FIG. 13. The kidney, pancreas, and spleen were sectioned and IHC-stained with anti-human CD45 to detect T cell therapies as in FIG. 13. For FIGs. 23A-23F, scale bars in IHC micrographs are 500 pm. FIG. 23G shows human CD45+ cell numbers were obtained as described herein. Tissue area was computationally obtained. Organ and tumor infiltration of human CD45+ cells per area were calculated and plotted. For calculation of cells / cm2in organs, n = 6 for NTD or n = 8 for VR-M5CARs, with 2 non-consecutive tissue slides stained per mouse and 3-4 mice per group; n = 7 for M5CAR alone, with 2 non-consecutive tissue slides stained for 3 mice and 1 tissue slide stained for 1 mouse. For calculation of cells / cm2in tumors, n = 6 for NTD or n = 8 for M5CAR alone and VR-M5CARs, with 2 non-consecutive tissue slides stained per mouse and 3-4 mice per group. FIG. 23H shows mouse body weights were collected at the end of the study. Body weights are plotted as mean ± SEM (n = 5 mice per group, except for PBS for which n = 2, NTD for which n = 3, and M5CAR alone for which n = 4). Ordinary one- way ANOVA with Tukey’s multiple comparison test was used for statistical analysis (ns = not significant).

[0046] FIGs. 24A-24C show 115, ZL8, TNFa, and IFNg secretion by mesothelin-expressing ASPC1 (FIG. 24A), H226 (FIG. 24B), and 0VCAR3 cells (FIG. 24C) after overnight incubation at increasing concentrations of 2.5e4 cells / mL, 5e4 cells / mL. and le5 cells / mL (from left to right). FIGs. 24D- 24S show 115, IL8, TNFa, and IFNg secretion by M5CAR T cells and VR-M5CAR T cells from two donors after overnight co-incubation with mesothelin-expressing ASPC1, H226, and 0VCAR3 cells or incubation in media alone, as indicated in the figure. Cytokine release was measured using ELISA. Mean with SEM is shown (n = 3). One-way ANOVA with Tukey’s multiple comparison test was used for statistical analysis (ns = not significant, *P < 0.05, **P < 0 0!, ***P <0.001, ****P <0.0001).

[0047] FIGs. 25A-25B show surface expression of CD4 and CD8 on two donors, as determined by flow cytometry using BV421 anti-CD4 and PE anti-CD8. FIGs. 25C-25D show surface expression of the activation markers CD69 and CD25 on two donors, as determined by flow cytometry using BV421 anti-CD69 and PE anti-CD25. FIGs. 25E-25F show surface expression of the exhaustion markers CTLA4 and PD1 on two donors, as determined by flow cytometry using BV421 anti- CTLA4 and PE anti-PDl . Surface expression was analyzed in FlowJo on M5CAR+-gated T cells as determined by GFP expression.

[0048] FIGs. 26A-26B show surface expression of the memory markers CD45RA and CCR7 on two donors, as determined by flow cytometry using BV421 anti-CD45RA and APC anti-CCR7. FIGs. 26C-26D show surface expression of the Treg markers CD 127 and CD 152 on two donors, as determined by flow cytometry using V50 anti-CD127 and BB700 anti-CD152. Surface expression was analyzed in FlowJo on M5CAR+-gated T cells as determined by GFP expression.

[0049] DETAILED DESCRIPTION

[0050] Provided herein are chimeric polypeptides that include (a) an extracellular domain comprising a cytokine-binding domain; (b) an extracellular hinge domain; (c) a transmembrane domain; and (d) an intracellular signaling domain. In some embodiments, the chimeric polypeptides described herein can be used to produce velocity receptor (VR) expressing immune cells (e.g., T cells) for the treatment of disease (e.g., cancer). Also provided herein are nucleic acids encoding any of the variety of the chimeric polypeptides described herein. Also provided herein are recombinant vectors, immune cells, or pharmaceutical compositions that comprise any of the variety of the nucleic acids described herein.

[0051] Also provided herein are methods of treating a subject, the method including administering to the subject any of the variety of the pharmaceutical compositions described herein. Also provided herein are methods of modulating immune cell migration, such methods including delivering any of the variety of the chimeric polypeptides, nucleic acids, or recombinant vectors into an immune cell.

[0052] Various non-limiting aspects of these chimeric polypeptides are described herein, and can be used in any combination without limitation. Additional aspects of various components of methods of making and using the chimeric polypeptides are known in the art.

[0053] As used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise.

[0054] As used herein, the term “about”, when used herein in reference to a value, refers to a value that is similar, in context to the referenced value. In general, those skilled in the art, familiar with the context, will appreciate the relevant degree of variance encompassed by “about” in that context. For example, in some embodiments, the term “about” may encompass a range of values that are within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the referred value.

[0055] As used herein, the term “administration” typically refers to the administration of a composition to a subject or system to achieve delivery of an agent that is, or is included in, the composition. Those of ordinary skill in the art will be aware of a variety of routes that may, in appropriate circumstances, be utilized for administration to a subject, for example a human. For example, in some embodiments, administration may be intra-arterial, intravenous, intraventricular, or within a specific organ (e. g., intrahepatic). In some embodiments, administration may involve only a single dose. In some embodiments, administration may involve application of a fixed number of doses. In some embodiments, administration may involve dosing that is intermittent (e.g., a plurality of doses separated in time) and / or periodic (e.g., individual doses separated by a common period of time) dosing. In some embodiments, administration may involve continuous dosing (e g., perfusion) for at least a selected period of time. 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 any of a variety of contexts - including where interacting entities or moieties are studied in isolation or 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).

[0056] As used herein, in general, the term “engineered” refers to the aspect of having been manipulated by the hand of man. For example, a polypeptide is considered to be “engineered” when the polypeptide sequence manipulated by the hand of man. For example, in some embodiments of the present invention, an engineered polypeptide comprises a sequence that includes one or more amino acid mutations, deletions, and / or insertions that have been introduced by the hand of man into a reference polypeptide sequence. In some embodiments, an engineered polypeptide includes a polypeptide that has been fused (i.e., covalently linked) to one or more additional polypeptides by the hand of man, to form a fusion polypeptide that would not naturally occur in vivo. Comparably, a cell or organism is considered to be “engineered” if it has been manipulated 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). As is common practice and is understood by those in the art, derivatives and / or progeny of an engineered polypeptide or cell are typically still referred to as “engineered” even though the actual manipulation was performed on a prior entity.

[0057] As used herein, the term “pharmaceutical composition” refers to a composition in which an active agent is formulated together with one or more pharmaceutically acceptable carriers. In some embodiments, the composition is suitable for administration to a human or animal subject. In some embodiments, the active agent is present in unit dose amount appropriate for administration in a therapeutic regimen that shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population.

[0058] As used herein, the term “specific binding” refers to an ability to discriminate between possible binding partners in the environment in which binding is to occur. A binding agent that interacts with one particular target when other potential targets are present is said to “bind specifically” to the target with which it interacts. In some embodiments, specific binding is assessed by detecting or determining degree of association between the binding agent and its partner; in some embodiments, specific binding is assessed by detecting or determining degree of dissociation of a binding agent-partner complex; in some embodiments, specific binding is assessed by detecting or determining ability of the binding agent to compete an alternative interaction between its partner and another entity. In some embodiments, specific binding is assessed by performing such detections or determinations across a range of concentrations.

[0059] As used herein, the term “subject” refers an organism, typically a mammal (e.g., a human). In some embodiments, a subject is suffering from a relevant disease, disorder or condition. In some embodiments, a subject is susceptible to a disease, disorder, or condition. In some embodiments, a subject displays one or more symptoms or characteristics of a disease, disorder, or condition. In some embodiments, a subject does not display any symptom or characteristic of a disease, disorder, or condition. In some embodiments, a subject is someone with one or more features characteristic of susceptibility to or risk of a disease, disorder, or condition. In some embodiments, a subject is a patient. In some embodiments, a subject is an individual to whom diagnosis and / or therapy is and / or has been administered.

[0060] As used herein, a “vector” or “recombinant vector” refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a “plasmid”, which refers to a circular double stranded DNA loop into which additional DNA segments may be ligated. Another type of vector is a viral vector, wherein additional DNA segments may be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) can be integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as “expression vectors.” Standard techniques may be used for recombinant DNA, oligonucleotide synthesis, tissue culture, and transformation (e.g., electroporation, lipofection). Enzymatic reactions and purification techniques may be performed according to manufacturer's specifications or as commonly accomplished in the art or as described herein. The foregoing techniques and procedures may be generally performed according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification. See e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual 2nded., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989)), which is incorporated herein by reference for any purpose.

[0061] Chimeric Polypeptides

[0062] Provided herein are chimeric polypeptides comprising (a) an extracellular domain comprising a cytokine-binding domain; (b) an extracellular hinge domain; (c) a transmembrane domain; and (d) an intracellular signaling domain. In some embodiments, a chimeric polypeptide can include (a) an extracellular domain comprising a cytokine-binding domain; (b) an extracellular hinge domain; (c) a transmembrane domain; (d) an intracellular signaling domain; or any combinations thereof.

[0063] Chimeric polypeptides can comprise a single polypeptide chain or multiple polypeptide chains. In some embodiments, a chimeric polypeptide comprises a single polypeptide. In some embodiments, a chimeric polypeptide comprises two polypeptides. In some embodiments, a chimeric polypeptide comprises more than two polypeptides.

[0064] In some embodiments, a single polypeptide of the chimeric polypeptide can include amino acid sequences (e.g., domains) derived from at least two different sources (e.g., two different naturally-occurring proteins). In some embodiments, a single polypeptide of the chimeric polypeptide can include domains from at least two different naturally-occurring animal proteins. In some embodiments, a single polypeptide of the chimeric polypeptide can include domains from at least two different naturally-occurring mammalian proteins. In some embodiments, a single polypeptide of the chimeric polypeptide can include domains from at least two different naturally- occurring human proteins. In some embodiments, a single polypeptide of the chimeric polypeptide can include a domain that is a synthetic sequence (e.g., a scFv) and a domain that is derived from a naturally-occurring protein (e.g., a naturally-occurring animal or human protein). In some embodiments, a single polypeptide of the chimeric polypeptide can include at least two different domains that are synthetic sequences (e.g., two different scFvs; one scFv and another synthetic sequence).

[0065] In some embodiments, a chimeric polypeptide comprises two or more polypeptides, wherein the two or more polypeptides can include amino acid sequences (e.g., domains) derived from at least two different sources (e.g., two different naturally-occurring proteins). In some embodiments, the two or more polypeptides can include domains from at least two different naturally-occurring animal proteins. In some embodiments, the two or more polypeptides can include domains from at least two different naturally-occurring mammalian proteins. In some embodiments, the two or more polypeptides can include domains from at least two different naturally-occurring human proteins. In some embodiments, the two or more polypeptides can include a domain that is a synthetic sequence (e.g., a scFv) and a domain that is derived from a naturally-occurring protein (e g., a naturally-occurring animal or human protein). In some embodiments, the two or more polypeptides can include at least two different domains that are synthetic sequences (e.g., two different scFvs; one scFv and another synthetic sequence). In some embodiments, the two or more polypeptides can include a domain that is not a protein / peptide (e g., a nucleic acid, a lipid, a phospholipid, a glycan).

[0066] In some embodiments, the two or more polypeptides are reversibly linked. In some embodiments, the two or more polypeptides are irreversibly linked. In some embodiments, the two or more polypeptides are covalently linked. In some embodiments, the two or more polypeptides are non-covalently linked. In some embodiments, the two or more polypeptides are linked by dimerization or bioconjugation domains. In some embodiments, the chimeric polypeptide further comprises a cleaving sequence, wherein the cleaving sequence couples the two or more polypeptides. In some embodiments, the cleaving sequence comprises a P2A, T2A, E2A, F2A, or IRES cleaving sequence.

[0067] Velocity Receptor (VR) Expressing Immune Cells

[0068] In some embodiments, a chimeric polypeptide described herein is or includes a velocity receptor (VR). As used herein, a “velocity receptor” refers to a receptor that comprises a cytokinebinding domain (e.g., an scFv, the binding domain of a cytokine receptor), an extracellular domain that tethers the cytokine-binding domain to the plasma membrane, a transmembrane domain, and one or more cytokine receptor signaling domains. In some embodiments, a cytokine bound by the VR is a soluble cytokine, or cytokine bound to soluble or membrane-bound cytokine receptors. In some embodiments, the cytokine can modulate immune cell migration. In some embodiments, the cytokine can increase immune cell migration. In some embodiments, the cytokine can decrease immune cell migration. In some embodiments, a velocity receptor can be expressed on an immune cell, wherein the immune cell is a T cell, B cell, Natural Killer (NK) cell, dendritic cell (DC), macrophage, monocyte, eosinophil, basophil, neutrophil, mast cell, lymphoid progenitor cell, myeloid progenitor cell, hematopoietic stem cell, red blood cell, platelet, megakaryocyte, or their any of their subtypes.

[0069] As used herein, “immune cells” refer to cells of the immune system which can be categorized as lymphocytes (e.g., T cells, B cells, natural killer [NK] cells, NKT cells), dendritic cells, monocytes / macrophages, granulocytes (e.g., neutrophils, eosinophils, basophils), mast cells, and their subtypes. In some embodiments, the immune cell is a T cell. In specific embodiments, the immune cell can be a subset of T cells, e.g., a aP-T cell, a yb-T cell, a CD4+T cell, a CD8+T cell, a CD3+CD4-CD8- double-negative (DN) T cell, a Thl T cell, a Th2 T cell, a Th 17 T cell, a regulatory T cell (Treg), another subset of T cells, or a population of T cells that comprises a combination of any of the foregoing. In some embodiments, an immune cell is an engineered immune cell, which means the immune cell has been genetically modified to express a non- naturally occurring protein (e.g., velocity receptor), or modified to include a non-coded amino acid, or modified to include posttranslational modifications, or modified to include an exogenous nucleic acid.

[0070] In some embodiments, an immune cell expressing a VR exhibits increased chemotaxis (e.g., increased chemotaxis in response to a protein (e.g., a cytokine) that the VR recognizes). Additionally or alternatively, in some embodiments, an immune cell expressing a VR exhibits increased basal migration.

[0071] Cytokine -Binding Domain

[0072] In some embodiments, a chimeric polypeptide comprising a VR provided herein includes an extracellular domain comprising a cytokine-binding domain. In some embodiments, the cytokine-binding domain comprises an scFv. In some embodiments, the cytokine-binding domain comprises a human scFv. In some embodiments, the cytokine-binding domain comprises a mouse scFv. In some embodiments, the scFv specifically binds a cytokine selected from IL5, IL8, IFN- gamma, TNF-alpha, or any combination thereof. In some embodiments, the scFv comprises one or any combination of its light chain complementarity-determining regions (e.g., LC CDR1, LC CDR2, LC CDR3). In some embodiments, the scFv comprises one or any combination of its heavy chain complementarity-determining regions (e g., HC CDR1, HC CDR2, HC CDR3).

[0073] In some embodiments, a chimeric polypeptide comprising a VR comprises a single extracellular domain. In some embodiments, a chimeric polypeptide comprising a VR comprises two extracellular domains. In some embodiments, a chimeric polypeptide comprising a VR comprises three extracellular domains. In some embodiments, a chimeric polypeptide comprising a VR comprises four or more extracellular domains. In some embodiments, the cytokine-binding domain comprises an IL5R-alpha, IL5R-beta, IFN-gammaRl, IFN-gammaR2, TNFR1, TNFR2 cytokine-binding domain, or any combination thereof. In some embodiments, the cytokine-binding domain specifically binds to IL5, IL8, IFN-gamma, TNF-alpha, or any combination thereof.

[0074] In some embodiments, the cytokine-binding domain specifically binds to IL5, wherein the chimeric polypeptide comprises SEQ ID NO: 93, SEQ ID NO: 16, SEQ ID NO: 84, SEQ ID NO: 17, SEQ ID NO: 85, SEQ ID NO: 73, SEQ ID NO: 78, SEQ ID NO: 12, SEQ ID NO: 15, SEQ ID NO: 21, and SEQ ID NO: 24. In some embodiments, the cytokine-binding domain specifically binds to IL8, wherein the chimeric polypeptide comprises SEQ ID NO: 93, SEQ ID NO: 69, SEQ ID NO: 84, SEQ ID NO: 70, SEQ ID NO: 85, SEQ ID NO: 73, SEQ ID NO: 78, SEQ ID NO: 12, SEQ ID NO: 15, SEQ ID NO: 21, and SEQ ID NO: 24. In some embodiments, the cytokinebinding domain specifically binds to IFN-gamma, wherein the chimeric polypeptide comprises SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, and SEQ ID NO: 33. In some embodiments, the cytokine-binding domain specifically binds to TFN-alpha, wherein the chimeric polypeptide comprises SEQ ID NO: 93, SEQ ID NO: 197, SEQ ID NO: 84, SEQ ID NO: 198, SEQ ID NO: 85, SEQ ID NO: 73, SEQ ID NO: 78, SEQ ID NO: 12, SEQ ID NO: 15, SEQ ID NO: 21, and SEQ ID NO: 24.

[0075] Intracellular Signaling Domain

[0076] In some embodiments, a chimeric polypeptide comprising a VR described herein includes an intracellular signaling domain. In some embodiments, an N-terminus of the cytokine-binding domain is coupled to an intracellular signaling domain. In some embodiments, a chimeric polypeptide comprising a VR comprises a single intracellular signaling domain. In some embodiments, a chimeric polypeptide comprising a VR comprises two intracellular signaling domains. In some embodiments, a chimeric polypeptide comprising a VR comprises three intracellular signaling domains. In some embodiments, a chimeric polypeptide comprising a VR comprises four or more intracellular signaling domains. In some embodiments, the intracellular signaling domain is selected from IL5R-alpha, IL5R-beta, IL8R-alpha, IL8R-beta, IFN-gammaRl, IFN-gammaR2, TNFR1, TNFR2, CD8-alpha, CD28 signaling domain, or any combination thereof. In some embodiments, the cytokine-binding domain is coupled to the intracellular signaling domain, wherein the intracellular signaling domain comprises an IL5R-alpha, IL5R-beta, IL8R-alpha, IL8R-beta, IFN-gammaRl, IFN-gammaR2, TNFR1, TNFR2, CD8-alpha, CD28 intracellular signaling domain, or any combination thereof. In some embodiments, the intracellular signaling domain comprises an IL5R-alpha, IL5R-beta, IFN-gammaRl, IFN-gammaR2, TNFR1, TNFR2 intracellular signaling domain, one or any combination or all of the four intracellular domains of IL8R-alpha, or one or any combination or all of the four intracellular domains of IL8R- beta.

[0077] Hinge Domain

[0078] In some embodiments, a chimeric polypeptide described herein includes an extracellular hinge domain. In some embodiments, the hinge domain comprises an IL5R-alpha, IL5R-beta, IFN- gammaRl, IFN-gammaR2, TNFR1, TNFR2, CD8-alpha, CD4, CD28, IgGl, IgG4 hinge domain, one or any combination or all of the four hinge domains of IL8R-alpha, or one or any combination or all of the four hinge domains of IL8R-beta.

[0079] Transmembrane Domain

[0080] In some embodiments, a chimeric polypeptide described herein includes a transmembrane domain. In some embodiments, the transmembrane domain comprises an IL5R-alpha, IL5R-beta, IFN-gammaRl, IFN-gammaR2, TNFR1, TNFR2, CD8-alpha, CD28 transmembrane domain, one or any combination or all of the seven transmembrane domains of IL8R-alpha, or one or any combination or all of the seven transmembrane domains of IL8R-beta.

[0081] Nucleic acids and vectors Provided herein are nucleic acids encoding any of the variety of the chimeric polypeptides described herein. As used herein, “nucleic acid” is used to include any compound and / or substance that comprise a polymer of nucleotides. In some embodiments, a polymer of nucleotides is referred to as polynucleotides. Exemplary nucleic acids or polynucleotides can include, but are not limited to, ribonucleic acids (RNAs) and / or deoxyribonucleic acids (DNAs).

[0082] In some embodiments, nucleic acid constructs may be inserted into a recombinant vector or viral vector by methods known to the art, and nucleic acid molecules may be operably linked to an expression control sequence. Non-limiting examples of recombinant vectors include plasmid vectors, transposon vectors, cosmid vectors, and viral vectors (e.g., any adenoviral vectors, cytomegaloviral [CMV] vectors, simian viral [SV40] vectors, adeno-associated virus vectors, lentiviral vectors, and retroviral vectors). In some embodiments, the recombinant vector is a viral vector.

[0083] Additional sequences can be added to such cloning and / or expression sequences to optimize their function in cloning and / or expression, to aid in isolation of the polynucleotide, or to improve the introduction of the polynucleotide into a cell. Use of cloning vectors, recombinant vectors, adapters, and linkers is well known in the art.

[0084] Also provided herein are recombinant vectors comprising any of the variety of the nucleic acid molecules or comprising nucleic acids encoding for any of the amino acid sequences described herein. In some embodiments, nucleic acid constructs include regions that encode a velocity receptor (VR). In some embodiments, the recombinant vector further comprises a promoter operationally linked to the nucleic acid. In some embodiments, the promoter is a constitutive promoter. In some embodiments, the promoter is an inducible promoter. In some embodiments, the promoter comprises an EFl -alpha, CMV, CAG, PgK, or SV40 promoter. In some embodiments, the recombinant vector is a viral vector. In some embodiments, the viral vector is a lentiviral vector.

[0085] Also provided herein are immune cells comprising any of the variety of the nucleic acid molecules or the vectors described herein. In some embodiments, nucleic acid molecules are inserted into a vector that is able to express a chimeric polypeptide of the present disclosure when introduced into an appropriate cell. Methods of introducing nucleic acids and expression vectors into a cell (e.g., an immune cell) are known in the art. Non-limiting examples of methods that can be used to introduce a nucleic acid into a cell include electroporation, microinjection, any form of transfection, lipofection, calcium phosphate transfection, dendrimer-based transfection, cationic polymer transfection, cell squeezing, sonoporation, optical transfection, impalefection, hydrodynamic delivery, magnetofection, nanoparticle transfection, cell-penetrating peptides, or viral transduction.

[0086] In some embodiments, a nucleic acid encoding a chimeric polypeptide described herein can be delivered to cells by methods that include one or any combination of packaging the nucleic acid in a lentivirus, retrovirus, AAV, adenovirus, or delivery using CRISPR, lipid drug delivery nanotechnology, micelle drug delivery nanotechnology, polymer drug delivery nanotechnology, or mRNA technology.

[0087] In some embodiments, a nucleic acid encoding a chimeric polypeptide described herein can be cloned into a plasmid under the control of overexpression promoters (e.g., EFl-alpha, CMV, CAG, PgK, or SV40). In some embodiments, the plasmid backbone is a 1st, 2nd, or 3rd generation lentivirus backbone.

[0088] In some embodiments, a nucleic acid encoding a chimeric polypeptide described herein can be incorporated into the same backbone in any possible combination and ordering. In some embodiments, the nucleic acids that are incorporated into the same backbone can be separated by cleaving sequences that translate into cleaving peptides (e.g., P2A, T2A, E2A, F2A, or IRES).

[0089] In some embodiments, a nucleic acid encoding a chimeric polypeptide described herein can be incorporated into the same backbone in any possible combination and ordering with nucleic acids encoding receptors or molecules that enhance immune cell function against disease (e.g., CARs, TCR technologies, BCR technologies, immune suppression receptors and soluble molecules, immune activators and soluble molecules, exhaustion-specific receptors and soluble molecules).

[0090] Therapeutic applications

[0091] Provided herein are pharmaceutical compositions that include any of the variety of chimeric polypeptides, the nucleic acid molecules, the recombinant vectors, or the immune cells described herein. In some embodiments, pharmaceutical compositions provided herein include a pharmaceutically acceptable carrier.

[0092] Also provided herein are methods of treating a disease or condition in a subject that include administering to the subject any one of the chimeric polypeptides, the nucleic acid molecules, the recombinant vectors, or the immune cells described herein, or the pharmaceutical compositions described herein. In some embodiments, the therapeutic is administered as a polypeptide in its active form. In some embodiments, the therapeutic is administered as a polypeptide in an inactive form (e.g., proform). In some embodiments, the therapeutic is administered as a vector for expression in vivo.

[0093] In some embodiments, the disease or condition comprises cancer, autoimmune disease, wound healing, heart disease, or allergy. In some embodiments, the disease or condition comprises a solid cancer. In some embodiments, the disease or condition comprises neurodegenerative disease, kidney disease, or viral infection. In some embodiments, the disease or condition comprises diabetes, inflammation, genetic disorders, viral infections, bacterial infections, fibrosis, cardiovascular disease, kidney disease, lung disease, graft vs host disease, gastrointestinal disease, neurodegenerative disorders, osteoporosis and bone disease, digestive disease, skin disease, periodontal disease, dermatologic conditions, or cosmetic procedures.

[0094] Also provided herein are methods of modulating immune cell migration that include delivering any of the variety of the chimeric polypeptides, the nucleic acids, or the recombinant vectors into an immune cell. In some embodiments, the modulating comprises increasing immune cell migration. In some embodiments, the modulating comprises decreasing immune cell migration. In some embodiments, the immune cell is a human immune cell.

[0095] Pharmaceutical compositions

[0096] In some embodiments, the present disclosure provides pharmaceutical compositions that include any of the chimeric polypeptides, the nucleic acid molecules, the recombinant vectors, or the cells described herein, and a pharmaceutically acceptable carrier. In some embodiments, a pharmaceutical composition can include a buffer, a diluent, solubilizer, emulsifier, preservative, adjuvant, an excipient, or any combination thereof. In some embodiments, a composition, if desired, can also contain one or more additional therapeutically active substances.

[0097] In some embodiments, compositions are formulated for parenteral administration. For example, a pharmaceutical composition provided herein may be provided in a sterile injectable form (e.g., a form that is suitable for subcutaneous injection, intramuscular injection, or intravenous infusion). For example, in some embodiments, a pharmaceutical composition is provided in a liquid dosage form that is suitable for injection. In some embodiments, a pharmaceutical composition is provided as powders (e.g., lyophilized and / or sterilized), optionally under vacuum, which can be reconstituted with an aqueous diluent (e.g., water, buffer, salt solution, etc.) prior to injection. In some embodiments, a pharmaceutical composition is diluted and / or reconstituted in water, sodium chloride solution, sodium acetate solution, benzyl alcohol solution, phosphate buffered saline, etc. In some embodiments, a powder should be mixed gently with the aqueous diluent (e.g., not shaken).

[0098] In some embodiments, a pharmaceutical composition of the present disclosure is formulated with a pharmaceutically acceptable parenteral vehicle. Examples of such vehicles are water, saline, Ringer’s solution, dextrose solution, and 1-10% human serum albumin. Liposomes and nonaqueous vehicles such as fixed oils can also be used. A vehicle or lyophilized powder can contain additives that maintain isotonicity (e.g., sodium chloride, mannitol) and chemical stability (e.g., buffers and preservatives). In some embodiments, a formulation is sterilized by known or suitable techniques. A pharmaceutical composition may additionally comprise a pharmaceutically acceptable excipient, which, as used herein, includes any and all solvents, dispersion media, diluents, or other liquid vehicles, dispersion or suspension aids, surface active agents, isotonic agents, thickening, or emulsifying agents, preservatives, solid binders, lubricants and the like, as suited to the particular dosage form desired. Remington’s The Science and Practice of Pharmacy, 21st Edition, A. R. Gennaro (Lippincott, Williams & Wilkins, Baltimore, MD, 2006) discloses various excipients used in formulating pharmaceutical compositions and known techniques for the preparation thereof. Except insofar as any conventional excipient medium is incompatible with a substance or its derivatives, such as by producing any undesirable biological effect or otherwise interacting in a deleterious manner with any other component(s) of the pharmaceutical composition, its use is contemplated to be within the scope of this disclosure.

[0099] EXAMPLES

[0100] The disclosure is further described in the following examples, which do not limit the scope of the disclosure described in the claims.

[0101] Example 1 - Secretomics Analysis

[0102] T cells migrate under two main modes of migration: chemotaxis and basal migration. Chemotaxis is the directional migration of cells in response to a gradient of proteins that act as migratory stimulants in the direction of increasing volumetric concentrations of said proteins, i.e., chemokines. Basal migration is the random base migration of cells that has zero to small amounts of directionality. It was discovered that T cells migrate in a density-dependent fashion where an increase in T cell density is positively correlated with the migration speeds of T cells and the percentage of T cells that migrate distances more than the square of their own radii. When the media of cells at high density (HD) were placed on cells at low density (LD) and their migration was tracked through time-lapse microscopy and subsequent trajectory-mapping, it followed that LD cells increased their migration speeds and percent of the population’s motility.

[0103] Secretomics analysis revealed that this density-dependence motility is due to the increased secretion of, in part, the cytokines IL5, IL8, TNF-alpha, and IFN-gamma by T cells at high density (le6cells / ml) when compared to T cells at low density (le5cells / ml). In some embodiments, using the secretomics analysis method and comparison of cells at any densities deemed high and low densities, intracellular molecules that govern immune cell migration (e.g., IL5, IL8, TNF-alpha, and IFN-gamma) were found. Furthermore, the method described herein also showed not only the molecules found through this analysis, but their cognate receptors, the signaling components of these receptors, and the downstream signaling of these receptors, including but not limited to JAKs, STATs, TRADDs, TRAFFs, nFkB, actin assembly and depolymerization, myosin contractility, actin dynamics, myosin dynamics, microtubule dynamics, intermediate filament dynamics. It was shown that the motility of T cells at high density can be hampered if the interaction between these cytokines and their cognate receptors are blocked through neutralizing antibodies targeting the soluble cytokines or their membrane bound receptors, and also if downstream signaling molecules are blocked. It follows that the migration of these cells at low density can be enhanced by exogenously treating LD cells with any of the four cytokines IL5, IL8, TNF-alpha, and IFN- gamma.

[0104] Example 2 - Sequences of velocity receptors (VRs) that can be overexpressed to modulate immune cell migration

[0105] In one example of the present disclosure, the amino acid for a VR specific for IL5 consists of a binding domain taken from the binding domain of IL5R-alpha, a hinge domain taken from the hinge domain of IL5R-alpha, a transmembrane domain taken from the transmembrane domain of IL5R-alpha, and an intracellular domain taken from the intracellular domain of IL5R-alpha (SEQ ID NO: 1). The nucleic acid sequence is provided (SEQ ID NO: 2). In another example of the present disclosure, another amino acid for a VR specific for IL5 consists of a binding domain taken from the binding domain of IL5R-beta, a hinge domain taken from the hinge domain of IL5R- beta, a transmembrane domain taken from the transmembrane domain of IL5R- beta, and an intracellular domain taken from the intracellular domain of IL5R- beta (SEQ ID NO: 3). The nucleic acid sequence is provided (SEQ ID NO: 4). Another example of the present invention for the amino acid of a VR specific for IL5 consists of SEQ ID NO: 1 and SEQ ID NO: 3 separated by a cleaving sequence that is the self-cleaving peptide P2A (SEQ ID NO: 5), i.e. IL5R-alpha-P2A-IL5R-beta (SEQ ID NO: 6). The nucleic acid sequences for P2A and IL5R-alpha-P2A-IL5R-beta are provided (SEQ ID NO: 7, SEQ ID NO: 8). SEQ ID NO: 6 and its nucleic acid sequence (SEQ ID NO: 8) encode for a VR termed V5.

[0106] In one example of V5’s use, V5 was produced through synthetic nucleic acid production using the amino acid sequence for the various domains obtained from UniProt. V5 was then cloned through Gibson assembly into a 2ndgeneration lentiviral backbone under the control of the EF1- alpha promoter. This was termed the V5 plasmid, which was packaged into lentiviral particles using the packaging cell line HEK-293T. This was termed the V5 lentiviral solution, which was concentrated at a titer above 106IFU / mL. The V5 lentiviral solution was used as a combination therapy with a mesothelin-specific CAR. T cells were transduced with a cocktail of the V5 lentiviral solution and a lentiviral solution containing lentiviruses enclosing a plasmid for the expression of a mesothelin-specific CAR and eGFP connected by P2A at an MOI between 5-10 for each. V5 expression and CAR co-expression were determined through flow cytometry by IL5R-alpha and / or IL5R-beta staining for V5, and eGFP expression for the CAR. T cells were either used directly in functional studies or first underwent bead isolation for IL5R-beta since although P2A’s inclusion between IL5R-alpha and IL5R-beta is assumed to result in equimolar amounts of each receptor, it results in less amounts of what follows it (i.e., IL5R-beta) in reality. Hence, bead isolation for IL5R-beta was assumed to result in the increased purity of cells with equimolar expression of IL5R-beta and IL5R-alpha. If bead isolation was performed, flow cytometry was subsequently performed to validate the increase in purity of IL5R-beta+CAR+cells. It was noted that whether bead isolation was performed or not, the percentage of CAR+were normalized based on eGFP expression before any functional study was undertaken. A standard 2-dimensional (2D) luciferase-based cytotoxicity assay was performed to determine efficiency of cell killing against eGFP-luciferase expressing Raji lymphoma cancer cells. It was unexpected to measure increased cancer-cell killing of CAR-T cells that express V5s as its initial purpose was to increase CAR-T cell migration (FIG. 1). A standard 2D assay was not expected to provide results about motility as the therapies didn’t have to migrate to get to their target; after sedimenting to the bottom of the assay microwell they were in immediate contact with their target and as long as they recognized their target they elicited a cytotoxic immune response. It was noted that 2D was the appropriate format to test the cellular therapies against liquid tumors, as Raji cells were, since it most resembled and bore an example of the practical interaction between the two in patients with blood cancers treated with an infusion of cellular therapies into the bloodstream where their interaction will be more or less immediate. CAR-T cells for solid tumors on the other hand could be assayed in a 3-dimensional (3D) microenvironment since this most closely resembles the practical application of cellular therapies infused in patients with solid tumors as the therapies must exit the bloodstream and traverse the complex extracellular matrix (ECM) microenvironment specifically in order to get to a solid tumor bulk in tissues prior to any binding and recognition to tumor cells; then the cellular therapies must migrate within and throughout the tumor bulk to clear the patient of said tumor. However, the standard 2D assay was performed to test V5-expressing CAR-T cells against luciferase-mCherry expressing 0VCAR3 ovarian cancer cells, a type of solid tumor cells. Even against 0VCAR3s, VR-expressing CAR-Ts exhibited more cytotoxicity compared to regular CAR-T cells (FIG. 2). A 3D luciferase assay, which was the perferred assay to determine effectiveness against solid tumors, was employed to determine how well CAR-T cells that express V5s migrate, infdtrate, and kill 0VCAR3 organoids in a collagen-rich 3D environment. V5s were adequate to make CAR-T cells more lethal in 3D (FIG. 3), whereas standard CAR-T cells exhibited no killing, not due to their inability to recognize and subsequently lyse tumor cells but because of their inability to effectively migrate through a collagen-rich 3D ECM to even get to the tumor bulk in the time-scale employed (18 days).

[0107] In another example, a 38-day in vivo NOD scid gamma (NSG) 0VCAR3 ovarian cancer mouse model was performed to test the effectiveness of five combination therapies with the mesothelin-specific M5 CAR in a preclinical setting (FIGs. 4-8). 39 female NSG mice were injected subcutaneously in the right flank with 5 x 106luciferase-mCherry 0VCAR3 cells in 100 pL 1 : 1 MatrigekPBS. For the duration of the experiment, tumors were measured at least once weekly using digital calipers. On day 7, tumors were palpable, after which mice were randomized into 8 control / treatment groups of 5 mice each, except the saline group which had 4 mice. Mice were injected twice, on days 8 and 27. On day 8, mice were injected intravenously with 200 pL of the control s / treatments in PBS in the tail vein. On day 27, mice were injected intraperitoneally with 100 pL of the control s / treatments in PBS. On both injection days, mice were injected with either one of three controls: saline (PBS), non-transduced T cells (NTD), or M5 CAR-T cells (M5CAR alone). Also on both injection days, mice were injected with either one of five combination treatments where T cells were first lentivirally transduced with one of the following five velocity receptors then 24 hours later lentivirally transduced with an M5 CAR: V5 (FIG. 4), VR5u IL5 (FIG. 5), VR5a IL8 (FIG. 6), Vysig(FIG. 7), VR5a TN Fa. (FIG. 8). It was noted that for both injections on days 8 and 27, 3 x 106NTD, M5CAR, or combination treatment cells were injected. For the day 8 injection, M5CAR alone and combination treatment cells were normalized to 45% positivity of the M5 CAR, whereas they were normalized to 71% for the day 27 injection. All combination treatments were better at controlling tumor growth than the T cells that were transduced with M5 CAR alone (FIGs. 4-8).

[0108] For the combination treatment groups, the velocity receptors are as follows. V5 is the same construct as previously described.

[0109] VR5a IL5 is a VR specific for IL5 which consists of, from 5’ to 3’, a signaling peptide taken from the signaling peptide of CD8a (SEQ ID NO: 93), a binding domain taken from the VH domain of an IL5-specific scFv (SEQ ID NO: 16), a linker (SEQ ID NO: 84), the VL domain of an IL5-specific scFv (SEQ ID NO: 17), an 8x His tag (SEQ ID NO: 85), linked to a hinge domain taken from the hinge domain of CD8a (SEQ ID NO: 73), a transmembrane domain taken from the transmembrane domain of CD8a (SEQ ID NO: 78), and an intracellular domain taken from the signaling domains of IL5R-alpha (SEQ ID NO: 12), IL5R-beta (SEQ ID NO: 15), TNFR1 (SEQ ID NO: 21), and TNFR2 (SEQ ID NO: 24).

[0110] VR5a IL8 is a VR specific for IL8 which consists of, from 5’ to 3’, a signaling peptide taken from the signaling peptide of CD8a (SEQ ID NO: 93), a binding domain taken from the VH domain of an IL8-specific scFv (SEQ ID NO: 69), a linker (SEQ ID NO: 84), the VL domain of an IL8-specific scFv (SEQ ID NO: 70), an 8x His tag (SEQ ID NO: 85), linked to a hinge domain taken from the hinge domain of CD8a (SEQ ID NO: 73), a transmembrane domain taken from the transmembrane domain of CD8a (SEQ ID NO: 78), and an intracellular domain taken from the signaling domains of IL5R-alpha (SEQ ID NO: 12), IL5R-beta (SEQ ID NO: 15), TNFR1 (SEQ ID NO: 21), and TNFR2 (SEQ ID NO: 24).

[0111] Vysig is a VR specific for IFNy which consists of, from 5’ to 3’, a binding and hinge domain taken from the binding and hinge domains of IFNGR1 (SEQ ID NO: 28), a transmembrane domain taken from the transmembrane domain of IFNGR1 (SEQ ID NO: 29), and an intracellular domain taken from the signaling domains of IFNGR1 (SEQ ID NO: 30) and IFNGR2 (SEQ ID NO: 33).

[0112] VR5a TNFa is a VR specific for TNFa which consists of, from 5’ to 3’, a signaling peptide taken from the signaling peptide of CD8a (SEQ ID NO: 93), a binding domain taken from the VH domain of an TNFa-specific scFv (SEQ ID NO: 197), a linker (SEQ ID NO: 84), the VL domain of an TNFa-specific scFv (SEQ ID NO: 198), an 8x His tag (SEQ ID NO: 85), linked to a hinge domain taken from the hinge domain of CD8a (SEQ ID NO: 73), a transmembrane domain taken from the transmembrane domain of CD8a (SEQ ID NO: 78), and an intracellular domain taken from the signaling domains of IL5R-alpha (SEQ ID NO: 12), IL5R-beta (SEQ ID NO: 15), TNFR1 (SEQ ID NO: 21), and TNFR2 (SEQ ID NO: 24).

[0113] Human cancer cells

[0114] OVCAR3 (high-grade ovarian serous adenocarcinoma), AsPCl (pancreatic ductal adenocarcinoma), H226 (squamous cell lung carcinoma) cell lines were obtained from ATCC. OVCAR3 cells were maintained in RPMI supplemented with 10% FBS, 1% penicillin / streptomycin, 200 mM L-glutamine, 1% HEPES, and 1% sodium pyruvate. AsPCl cells were maintained in RPMI supplemented with 20% FBS, 1% penicillin / streptomycin, and 1% HEPES. H226 cells were maintained in RPMI supplemented with 10% FBS and 1% penicillin / streptomycin. For in vitro co-culture cytotoxicity studies, all cell lines were lentivirally transduced to stably express luciferase-P2A-mCherry. Following transduction, all cell lines underwent FACS cell sorting using a Sony SH800 cell sorter.

[0115] Donor primary human T cells

[0116] Primary human T cells were obtained from fresh identity-unlinked leukapheresis packs (LPs) from healthy donors who underwent same-day apheresis through the Anne Arundel Medical Center (Annapolis, MD, USA), which resulted in fresh leukapheresis packs (LPs). Freshly delivered LPs were immediately processed to extract PBMCs using Ficoll density gradients. PBMCs subsequently underwent negative selection for the isolation of pan T cells, or CD4 or CD8 T cell subsets for CAR manufacturing using STEMCELL negative selection isolation kits. T cells were maintained in X-Vivo 15 media supplemented with 5% Human AB serum, 200 mM L-glutamine, and 1% penicillin / streptomycin - after which, they were referred to as complete X-Vivo. For density-dependent migration studies (FIGs. 9B-9S), T cells were cultured in serum-free Immunocult media (STEMCELL) supplemented with 100 lU / ml IL2.

[0117] Cellular therapy manufacturing

[0118] VR therapy vectors and the CAR therapy vector were first designed through Gibson Assembly molecular cloning (HiFi DNA Assembly Cloning Kit, NEB) using the backbone pSLCAR-CD19- BBz (Addgene). VR nucleic acids were synthetically constructed as indicated (FIG. 10A), and were cloned into the backbone excluding the backbone’s native CD 19 CAR-P2A-GFP sequence to select for positive cells downstream through the use of surface staining with FACS antibodies for VRs and subsequent detection by flow cytometry. The M5CAR nucleic acid sequence was cloned into the backbone prior to the P2A-GFP sequence to select positive cells downstream by GFP using flow cytometry. VR therapy lentiviral particles and CAR therapy lentiviral particles were obtained by transfecting 293T cells with vectors for VRs or the M5CAR.

[0119] 293T cells were obtained from ATCC and maintained in either DMEM supplemented with 10% FBS or Opti-MEM supplemented with 5% FBS. Transfection was performed using GeneJuice, (Millipore Sigma) and lentiviral particles were concentrated in Lenti-X Concentrator (Lonza). VR, CAR, and VR-CAR therapies were manufactured as follows. CD4 and CD8 T cells were isolated using STEMCELL negative selection kits and were mixed 1 : 1. They were then activated overnight by CD3 / CD28 Dynabeads (Invitrogen) in complete X-Vivo media supplemented with 100 lU / ml IL2 (Peprotech). For VR alone or M5CAR alone therapies, activated cells were subsequently lentivirally transduced overnight on Retronectin-coated plates, after which their positivity was checked by flow cytometry. For the VR-CAR cells, after overnight activation, T cells were transduced on Retronectin-coated plated with VR therapy lentivirus overnight and checked for VR positivity by flow as follows: V5 stained for IL5RA; Vysig stained for IFNyRl; VR5ot IL5, VR5ot IL8, and VR5oc TNFoc stained for F(ab)2. VR T cells were then transduced overnight on Retronectin-coated plates with M5CAR therapy lentivirus and checked for M5CAR positivity by flow cytometric detection of GFP. VR-M5CAR, M5CAR, or non-transduced T cells were expanded for a total of 11 days. Dynabeads were magnetically removed using Dynamags (Invitrogen) and frozen in liquid nitrogen or maintained in culture in complete X-Vivo media supplemented with 100 lU / ml IL2.

[0120] 3D migration

[0121] T cells in complete X-Vivo media supplemented with 100 lU / ml IL2 were suspended in a 2 mg / ml type I collagen (Corning) solution. For density-dependent migration studies, T cells in Immunocult media were activated with Immunocult CD2 / CD3 / CD28 expansion solution (STEMCELL) supplemented with 100 lU / ml IL2. T cells were then suspended in 2 mg / ml type I collagen (Coming) and a 0.22 pm-filtered reconstitution buffer that consisted of HEPES, HBSS, and sodium bicarbonate in milliQ-purified water. The T cell-collagen solution was solidified in a 37°C and 5% CO2 incubator for 1 hr. Immunocult media supplemented with 100 lU / ml IL2 was then added on top of the T cell-collagen gels. The gels were then incubated for 48 hrs and imaged for 1 hr on a Nikon Ti2 microscope equipped with a live-cell system maintained at 37°C and 5% CO2. Time-lapse images were constructed into time-lapse movies using Nikon Elements, and cell positions were tracked frame-by-frame to generate cell trajectories using Metamorph. Analysis of cell trajectories to pull out multiple parameters to characterize cellular motility was performed in Matlab (MathWorks).

[0122] T cell secretomics

[0123] LD and HD T cells were cultured in 3D collagen gels, as described above, for 48 hrs in a 37°C and 5% CO2 incubator. Conditioned medium was collected from the top of 3D T cell-collagen gels and fdtered through a 0.45 pm fdter. Filtered conditioned medium was added to Isoplexis microchips.

[0124] 2D in vitro cytotoxicity

[0125] For T cell cytotoxicity, luciferase-expressing solid tumor cell lines were incubated at 37°C and 5% CO2 overnight. Subsequently, T cells at indicated E:T ratios were added on top of the cancer cells and incubated overnight. BrightGlo solution (Promega) was then added to the wells and luminescence was read on a SpectraMax plate reader (Molecular Devices) to obtain relative light units (RLUs). Specific lysis was calculated using the RLUs as 100% * (Alive - Condition) / ( Alive- Dead). Flow cytometry immunophenotyping

[0126] For immunophenotyping of VR-transduced T cells, 50 pL overnight-transduced T cells were collected, washed three times at 4°C in cold FACS buffer (PBS supplemented with 5% FBS and 0.1% sodium azide), and surfaced stained with an anti-F(ab)2 (VR5oc IL5, VR5ocTNFcc, and VR5ocIL8), anti-IL5RA (V5), or anti-IFNyRl (Vysig) antibody in FACS buffer at 4°C for 30 min. Cells were then washed three times at 4°C in FACS buffer before analysis by flow cytometry. Data was collected using Sony SH800 and was analyzed using FlowJo (BD). For M5CAR detection, T cells were collected and washed three times in FACS buffer and analyzed for GFP expression using flow cytometry. For activation marker expression immunophenotyping, T cells were collected, washed three times at 4°C in cold FACS buffer, and surface stained with an anti-CD25 or anti-CD69 antibody at 4°C for 30 min. Cells were washed three times at 4°C in cold FACS buffer. Surface-stained T cells were then analyzed by flow cytometry using Sony SH800. All data were analyzed using FlowJo software.

[0127] Cytokine assays

[0128] To measure T cell activity, AsPCl, OVCAR3, or H226 cells were incubated in overnight at 3e3 cells / well in 96 well plates. The following day, NTD, M5CAR, or VR-M5CAR T cells in were added to cancer cells at 1 : 1 E:T ratios and incubated overnight. 100 pL of supernatant was then carefully removed from each wells using a multichannel pipette. Supernatants were analyzed for IL2, TNFa, or IFNy by ELISA (Invitrogen) according to the manufacturer’s protocol.

[0129] Pre-clinical mouse models

[0130] All animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of the Johns Hopkins University (Protocol MOE23E29) and were performed in an Association for the Assessment and Accreditation of Laboratory Animal Care (AALAC) accredited facility. The housing group was five at maximum for all mice groups. Mouse studies were performed in 8-12-week-old NOD-.scvc / IL2Rgammanull (NSG) female and male mice (Jackson Laboratory). AsPCl (2e6) or H226 (5e6) human cells were subcutaneously injected into the right flank of each mouse with an equal volume of Matrigel (BD) to PBS. After tumors became palpable at roughly 100 - 250 mm3, mice were injected by tail vein (once) with either 100 pl PBS or non -transduced T cells, M5CAR T cells, or the indicated VR-M5CAR T cell therapies in 100 pl PBS. Tumors were measured using digital calipers approximately twice per week and mice were weighed at the end of each experiment. Tumor volumes were calculated using the following formula: volume: (L x W2) x 0.5. Tumors and organs were harvested at the indicated days. Half of the tissues were embedded in paraffin for IHC staining and the other half was flash-frozen.

[0131] Survival Mouse Studies

[0132] The OVCAR3 (5e6) cancer cell line was subcutaneously injected into the right flank of female NSG mice. After tumors became palpable mice were injected by tail vein with the respective treatment. Mice received a second dose after 19 days intraperitoneally. Tumors were measured twice a week as described above. Mice were sacrificed when they reached 750 mm3, and tumors and organs were collected.

[0133] Immunohistochemistry (IHC)

[0134] Mouse tumors and organs were fixed in formalin and subsequently paraffin-embedded and sectioned. Paraffin-embedded sections were blindly stained for human CD45 to measure human T cell infiltration into tumors and organs by The Johns Hopkins Oncology Tissue Core. Histological slides were scanned using Hamamatsu NanoZoom er S210 scanner at a 20x magnification (0.5pm / pixel). Quantification was done by a computational biologist who was blinded to the data.

[0135] Immunohistochemistry quantifications

[0136] Scanned histological images were downsampled to lOx magnification (1 pm / pixel) and color deconvolution was used to extract the IHC CD45 positive locations to an independent image channel. Intensity peaks were detected on the isolated CD45+ channel and spatial coordinates were collected (x, and y coordinates). Assessment of tumor CD45+ cell infiltration was performed by manual masking of each individual tumor location.

[0137] Statistical Analysis

[0138] All statistical tests were performed using GraphPad Prism. All migration MSD analyses used either the nonparametric Mann-Whitney ( / -test for two groups, or for multiple comparisons the Kruskal- Wallis ANOVA with Dunn’s post-test. All other analyses were performed using either the student’s / -test for two groups, or the ordinary one-way ANOVA with Tukey’s post-test for comparison of multiple groups. The Log rank Mantel-Cox test was used for Kaplan-Meier survival curves.

[0139] Example 3 - T cells produce cytokines to trigger their own migration

[0140] In order to develop a highly motile CAR T cell, and to understand how normal T cells migrate in 3D collagen-rich settings in vitro, it was hypothesized that identifying basic biophysical principles of enhanced T cell migration in simple in vitro models could help the design of CAR T cells featuring much higher infiltration potential in vivo. Using time-resolved phase contrast microscopy, the spontaneous movements (no added chemokines or chemoattractants) of hundreds of unlabeled primary human T cells purified from healthy donors were tracked, activated by CD3 / CD28 beads, and fully embedded in 3D collagen matrices. This standard 3D collagen matrix model has been used for cell migration studies and serves here as a screening platform. The mean squared displacement (MSD) of each cell was computed from the trajectories of cells in the matrix (FIG. 14A) and used as the primary parameter to compare the motility of individual cells. Due to the non-Gaussian nature of the distribution of cellular movements, traditional migration parameters such as speed and persistence time / distance could not be extracted. T cells exhibited heterogeneous migratory patterns. For a given recording time (typically 1 h), some T cells migrated more than their own size, ~5 pm, as measured from phase contrast micrograph while others hardly moved at all, suggesting different migratory states.

[0141] Recent work has shown that cell density (number of single cells in a set volume) can modulate the migration of cancer cells in collagen matrices. Breast cancer cells and fibrosarcoma cells showed a small, ~1.5-fold increase in cell speed for a 10-fold increase in cell density. Here, the possibility that cell density could modulate the migration of T cells was explored (FIG. 9B). It was found that T cells increased their migration up to 4-fold, as cell density was increased 10- fold, from I O’ to 106cells / ml (FIG. 9C and FIGs. 14A-14B). Note that these T cell densities were not chosen to reproduce physiological densities; they were chosen to elicit a potential change in cell speed that would help in the downstream identification of molecules responsible for this change. These cell densities were also chosen to correspond to intercellular distances that were sufficiently large for direct cell-cell collisions to be rare. Importantly, the MSD of the fastest moving T cells (T cells that migrated more than the square of their radius) were similar across all tested densities (FIGs. 14C-14D). This suggested that T cell migration adopted two main states: a high-motility state corresponding to the same high MSD (> R2, where R = 5 pm is the T cell radius) or a low-motility state for which significantly fewer cells were motile. This indicated that the differences in population-averaged MSD for different seeding densities were reflective of the percentage of T cells that were effectively migrating (FIG. 9D). In other words, as cell density was increased by loading collagen matrices with increasing numbers of cells, the population- averaged MSD increased not because of a change in the type of migration of T cells, but due to a higher fraction of cells in the high-motility state.

[0142] Since a high number of intercellular collisions that could have caused the increased cell migration were not observed, it was hypothesized that this density-induced increase in cell migration was caused by secreted molecules. To test this hypothesis, the supernatant was transferred from cells at high density (HD) in the collagen matrix to cells at low density (LD) (FIG. 9E). The conditioned medium (CM) harvested from cells in HD was sufficient to increase the motility of T cells at LD to levels similar to T cells at HD (FIG. 9F and FIGs. 14E-14F). Once again, this increase in population-averaged MSD corresponded to a shift in the number of cells in the high-motility state (FIG. 9G and FIGs. 14G-14H). This result suggested that factors secreted by cells at HD increased the percentage of cells in a high motility state.

[0143] Next, it was sought to identify the factors secreted by cells at HD responsible for their increased motility. A microchip was utilized to run single-cell proteomics on the medium from LD cells and HD cells suspended in 3D collagen matrices (FIG. 9H). Four cytokines were identified - IL8, TNFoc, IFNy, and IL5 - that were secreted by HD cells but undetectable for LD cells (FIG. 91), suggesting their de novo production by T cells at a high cellular density. The protease granzyme B, which remodels the ECM, thereby allowing more rapid T cell motility, was also detected for HD cells and undetected for LD cells. But, given the aim to engineer receptors that induce T cell migration signaling, it was decided to not pursue granzyme B as it has no known cognate receptor. IL8 is a well-established T cell chemoattractant, but little is known about its effect on T cell migration (i.e. not chemotaxis).

[0144] To understand if and how each of these cytokines modulated T cell motility, each recombinant cytokine was exogenously added separately to LD T cells in 3D collagen gels (FIG. 9 J). All cytokines increased T cell motility (FIG. 9K and FIGs. 14I-14J) by increasing the fraction of cells in the high-motility state (FIG. 9L and FIGs. 14K-14L). Thus, all four cytokines - IL5, TNFa, IFNy, and IL8 - can act on their own to increase T cell migration. Next, the receptor / cytokine interactions of cytokines endogenously produced at detectable levels in HD T cells were inhibited: IL5, TNFa, IFNy, and TL8 (FIG. 9M). Blocking each receptor / cytokine interaction hampered T cell migration (FIG. 9N and FIGs. 14M-14P) and, correspondingly, shifted a larger fraction of T cells to a low motility state (FIG. 90 and FIGs. 14Q-14T). Since each of the cytokine receptors signal through JAK / STAT, JAK / STAT signaling in HD cells was inhibited (FIG. 9P), a marked decrease in T cell motility (FIG. 9Q and FIGs. 14U-14V) and a shift into the low-motility state (FIG. 9R and FIGs. 14W-14X) was observed. In sum, T cells at high density produce IL5, TNFa, IFNy, and IL8 to shift their population to a self-propelled high- motility state and subsequently increase their population-averaged migration. These cytokines can be referred to as “velocity cytokines” (VCs) (FIG. 9S). Below, these results are shown to lock-in T cells into a high-motility state.

[0145] Example 4 - Velocity receptors push CAR T cells to a self-propelled high-motility state

[0146] It was sought to take advantage of VCs in a therapeutic setting, wherein current cellular therapies such as CAR T suffer from inadequate infiltration into solid tumors. Thus, a migration strategy was developed to enhance the infiltration of CAR T cells into solid tumors. One CAR that has received clinical interest in recent years with clinical trials still underway is the mesothelin- specific M5 CAR (M5CAR), which has the potential to ameliorate outcomes of multiple types of cancer patients with solid tumors, such as ovarian, pancreatic, and lung among others (NCT03054298 and NCT03323944).

[0147] For an enhanced migration strategy, overexpressing VCs to elevate their secretion was ruled out, as three VCs - TNFa, IFNy, and IL8 - have been implicated in cytokine release syndrome (CRS), one of the most severe CAR T cell therapy-related toxicity. Thus, it was decided to make M5CAR T cells that were more sensitive to VCs expressed by T cells themselves or already present in their environment, through the overexpression of velocity receptors (VRs). These receptors bind VCs and signal through VC cognate receptor signaling domains (FIG. 10A). VR-CAR T cells would thus exploit VCs that they secrete themselves and become self-propelled highly motile cells.

[0148] Since there are several VCs and each VCs’ cognate receptor is a heterodimer (except for IL8 which binds to both CXCR1 and CXCR2, composed of distinct domains), the VR design was made modular. First, a VR was defined as composed of four domains taken from a pool of each VCs’ cognate heterodimer receptor domains - binding, hinge, transmembrane (TM), and signaling. Next, four VC-specific scFvs (one for each VC) were included in this pool of domains, and thus a CD8a hinge and CD8a TM domain to tether the scFv to the cell membrane. These were included in part to 1) allow for a single receptor in contrast to a heterodimer, and 2) yield a higher binding affinity for VCs than their respective cognate receptors. There were > 30,000 VRs that could be constructed (FIG. 15A). 32 distinct VRs were cloned and the 5 that were successfully cloned were chosen. Out of practicality, it was decided to move forward with the 5 VRs to test and compare in vitro and preclinically in vivo (FIG. 10A). The 5 VRs were defined as 5 distinct therapies. They were VR5otIL5, VR5aTNFoc, VR5aIL8, V5, and Vysig (FIG. 10A). VR5aIL5, VR5aTNFa, and VR5ocIL8 have the same hinge (CD8a), TM (CD8a), and signaling (IL5RA-IL5RB-TNFR1- TNFR2) domains (FIG. 10A and FIG. 15B). Their only difference is their binding domain. They are all scFvs, but VR5ocIL5’s is a receptor for IL5, VR5aTNFot’s for TNFoc, and VR5aIL8’s for IL8 (FIG. 10A). V5 is an overexpression of the IL5 receptor (FIG. 10B). Vysig has an IFNyRl binding, hinge, and TM domain, but its signaling domain is IFNyRl -IFNyR2 (FIG. 10A and FIG.

[0149] 15B)

[0150] Since the CAR constructs of all FDA-approved CAR therapies to date are delivered to T cells virally, to compare performance with standard CAR T therapies, the VR and M5CAR genes were delivered to T cells lentivirally. Sequential transduction was performed, as the VR and M5CAR would have been indistinguishable in 3 of the therapies in cell-surface flow cytometric detection (FIG. 15B). VRs were each cloned into a lentiviral plasmid and used to transduced T cells. VR expression was detected by antibodies using flow cytometry (FIGs. 15B and 16A). The M5CAR was cloned into a lentiviral plasmid to express M5CAR-P2A-eGFP for flow cytometry detection without antibody staining and was used to transduce VR-T cells after VR transduction expression was confirmed by flow cytometry (FIGs. 15B and 16B).

[0151] As VRs were engineered to shift cells to a high-motility state, the migration status of VR- CAR T cells were next characterized to determine if these cells indeed migrated more than control CAR T cells (FIG. 10B). It was found that all 5 VRs in the VR-M5CAR therapies increased the migration of M5CAR T cells at LD up to levels similar or higher than M5CAR T cells at HD (FIG. 10C and FIGs. 16C-16D), and shifted a higher fraction of LD M5CAR T cells to a high-motility state (FIG. 10D and FIGs. 16E-16F) It was determined if this observed increased migration was the result of ligand-independent tonic VR signaling in the 3 scFv-based VR-CAR therapies, or if it was caused by VC binding as intended. VC-VR binding was inhibited using (separately) anti- IL5, anti-TNFa, anti-IL8 function-blocking antibodies (FIG. 10E). With these inhibitors added, VR-M5CAR T cells exhibited hindered motility (FIG. 10F and FIGs. 16G-16J) and shifted to a low-motility state (FIG. 10G and FIGs. 16K-16P).

[0152] The activation status of VR-M5CAR T cells compared to M5CAR alone was assessed through the expression of standard activation markers CD25 and CD69. All therapies, except Vysig, had higher CD69 expression than M5CAR T cells alone (FIG. 10H). They had greater than 3-fold expression of CD25 than M5CAR alone (FIG. 101). Using a standard overnight cell-killing assay, it was observed that VR-M5CAR cells were cytotoxic against multiple mesothelin-expressing solid cancer cells in vitro at similar levels to or greater levels than M5CAR T cells (FIG. 10J). VR-M5CAR cells were also either more active than or similarly active to M5CAR T cells, as determined by IL2, TNFoc, and IFNy release in response to overnight co-incubation with multiple mesothelin-expressing solid tumor cells in vitro (FIG. 10K).

[0153] These results demonstrate that VRs “lock in” a self-propelled high-motility state in CAR T cells. It was observed that VRs also have the added benefit of making CARs more activated, cytotoxic, and elicit stronger responses in the form of IL2, TNFa, and IFNy production against multiple solid cancers in vitro. These results suggested that the performance of VR-M5CAR therapies in preclinical mouse models could exceed that of M5CAR T cell therapy.

[0154] Example 5 - VR-M5CAR therapies infiltrate solid tumors in a pancreatic cancer preclinical model

[0155] To allow for the most meaningful comparison of the proposed VR-CAR T cells using current CAR T therapy, the mesothelin-specific CAR construct commonly used previously and targeted the same types of cancers was used. In the M5CAR phase 1 study, dose-limiting toxicities were observed with ineffective tumor infiltration. Likewise, other solid tumor targeting CAR T trials also observed dose limiting toxicities with inadequate tumor accumulation. T cell adoptive therapies in general have been infused preclinically in vivo in many cases at doses above 5 x 106T cells and sometimes with multiple administrations of such doses. Through standard intravenous (IV) administration at a dose where M5CAR T cells alone are ineffective at infiltrating tumors, it was hypothesized that migration strategies could not only help transport M5CAR T cells to the tumor site, but also increase their tumor-infiltration capabilities. It was decided to test the therapies in cancers included in the phase 1 study. First, pancreatic cancer was selected, which is defined as an immune-excluded tumor, where T cells generally form a ring around the tumor in the surrounding stroma with inefficient penetration into the tumor core in patients and animal models. Using the subcutaneous (s.c.) AsPCl NSG mouse model of pancreatic cancer extensively used in previous CAR T studies, we evaluated and compared the degree of cell infiltration was evaluated and compared and tumor growth was assessed. Cancer cells were subcutaneously injected into the right flank of the mouse. The tumors were allowed to become palpable (around 100-250 mm3), before treating the mice with the respective treatments. For treatments, the dose was lowered to one where CAR therapy alone was unable to infiltrate tumors and control tumor growth, i.e. 3 x 106cells in a single dose (i.v. injection), instead of a higher dose sometimes given multiple times during a treatment regimen.

[0156] Harkening back to what was observed in the phase 1 study, control M5CAR T cells (no VRs) were detected at low levels within tumors and often surrounding the tumor (FIG. 11A and FIGs. 17A-17C)). In contrast, even at this ineffective dose, it was found that several VR-M5CAR T cells infiltrated tumor cores readily and inhibited tumor growth. Not all VRs mediated a high number of M5CARs into tumors. The CAR T cells expressing VR5ocTNFa or VR5ocIL8 showed similar responses to the CAR alone (FIGs. 11B-11C and FIGs. 17D-17E). The general pancreatic cancer T cell landscape was observed in which T cells form a “ring” around tumors for M5CAR alone and for VR5alL8-M5CAR and Vysig-M5CAR cells, as they accumulated at the tumor edge; yet Vysig-M5CAR cells were detectable in the tumor core to a higher degree (FIG. 11D and FIG. 17F). Both VR5aIL5-M5CAR and V5-M5CAR cells infiltrated the entire tumor, edge and core, in large amounts (FIGs. 11E-11F and FIGs. 17G-17H). In addition, both VR5ocIL5-M5CAR and V5-M5CAR inhibited tumor growth (FIGs. 11E-11F and FIGs. 17G-17H). These qualitative assessments of CAR T cell infiltration were quantified using image analysis at single-cell resolution and normalized by the size of the tumor (FIG. 11G). For the best treatment, V5-M5CAR, tumor infiltration was increased by >10-fold compared to control M5CAR cells. This is an underestimate of the effect of V5 expression on CAR T infiltration, as the majority of M5CAR cells congregated at the tumor periphery. The volume of each tumor was plotted at the endpoint (FIGs. 11A-11F) as a function of the number of cells that had infiltrated that tumor (FIG. 11G) and found a remarkable inverse correlation (FIG. 11H): more CAR T cell infiltration mediated greater inhibition of tumor growth. VRs’ inhibition of tumor growth was validated using T cells from an additional donor.

[0157] The average volume of each tumor was plotted at the endpoint (FIGs. 11A-11F) as a function of the average number of cells that had infdtrated that tumor (FIG. 11G) and found a remarkable inverse correlation (FIG. 11H) more CAR T cell infdtration mediated greater inhibition of tumor growth, which supports our original hypothesis. The inhibition of tumor growth mediated by VR-M5CAR T cells was validated using T cells from an additional donor (FIG. 19).

[0158] Potential off-tumor effects of control M5CAR T cells and VR-M5CAR cells were assessed by measuring the human T cell burden in non-targeted organs, including the kidney and pancreas. Also, CAR detection was measured in the spleen, which is a therapeutically beneficial indication of CAR expansion. It was found that VRs had undetectable to minimal infiltration into the kidney and pancreas; far less than their accumulation in tumors (FIG. 20A-20G). VR5ocIL5-M5CAR and V5-M5CAR cells were detected in the spleen at far higher levels than other treatments (FIGs. 20A-20F). Moreover, no statistically significant loss of weight was observed in mice undergoing VR-M5CAR therapies (FIG. 20H).

[0159] Example 6 - VR-M5CAR therapies infiltrate solid tumors in a lung cancer preclinical model

[0160] The generality of the approach was tested by also testing VR-CAR therapies in a preclinical tumor model of lung cancer, as this was another cancer for which patients were recruited for the M5CAR phase 1 trial. For the same ineffective dose as in the pancreatic cancer model, control M5CAR cells were not seen to accumulate in either the tumor or in the surrounding stroma (FIG. 12A and FIGs. 21A-21C)). Co-transduction with VR5aTNFot showed similar responses to M5CAR alone (FIG. 12B and FIG. 21D) Vysig-M5CAR, VR5otIL8-M5CAR, and V5-M5CAR each formed a ring around tumor cores and led to similar tumor growth responses as M5CAR cells alone (FIGs. 12C-12E and FIGs. 21E-21G). VR5otIL5-M5CAR therapy had the highest accumulation within tumors, which correlated with attenuated tumor growth (FIG. 12F and FIG. 21H). CAR T cell infdtration was quantified at single-cell resolution and normalized by the size of the tumor (FIG. 12G) For the best treatment, VR5aIL5-M5CAR, tumor infiltration was increased by >10, OOO-fold compared to control M5CAR T cells. The volume of each tumor at the endpoint (FIGs. 12A-12F) was plotted as a function of the number of cells that had infiltrated that tumor (FIG. 12G). A remarkable inverse correlation was found (FIG. 12H): more CAR T cell infiltration mediated greater tumor growth inhibition. VRs’ ability to inhibit tumor growth was validated using T cells from an additional donor (FIGs. 22A-22G). Off-tumor effects were assessed for the kidney and pancreas. Accumulation of VR-M5CAR T cells were either nondetectable or miniscule (FIGs. 23A-23F). Off-tumor effects were assessed for the kidney and pancreas. Accumulation of combinations with VRs was either nondetectable or miniscule (FIGs. 23A-23F), especially when compared to tumor infiltration (FIG. 23G). VR5aIL5-M5CAR and V5-M5CAR showed infiltration into the spleen in greater amounts than other treatments (FIGs. 23A-23F). No significant loss of weight was observed in VR-M5CAR therapies compared to M5CAR alone (FIG. 23H).

[0161] As for pancreatic cancer (FIGs. 11B-11F), a general correlation was found between the degree of infiltration of these different engineered T cells in lung tumor and the growth inhibition of that tumor (FIGs. 12B-12F), which supports the hypothesis of an increased-migration strategy to enhance infiltration and associated therapeutic efficacy (FIG. 12G and 12H).

[0162] Example 7 - VR-M5CAR therapies increase overall survival in an ovarian cancer preclinical model

[0163] Since it was established that VRs can help M5CARs accumulate within multiple solid tumors and inhibit tumor growth, it was investigated if this translated into increased overall survival. To test this, an ovarian cancer preclinical model was utilized (FIG. 13A), as ovarian cancer was another cancer for which patients were recruited in the M5CAR phase 1 trial. Again, the ineffective M5CAR alone dose was kept and saw that each VR-M5CAR therapy was able to increase survival to a greater extent when compared to M5CAR alone (FIG. 18C). VR5aTNFa- M5CAR led to 20% overall survival after > 70 days (FIG. 13B). Co-transduction with Vysig or V5 led to longer survival compared to M5CAR alone (FIGs. 13C and 13D). Both VR5aIL8- M5CAR and VR5aIL5-M5CAR therapies each led to a months-long 100% overall survival (FIGs. 13E and 13F). Compared to M5CAR alone, no significant weight loss was observed in mice treated with VR-M5CAR T cells (FIG. 18D).

[0164] It was concluded that VR-M5CAR therapies designed to increase the migration of CAR T cells in a self-propelled manner inside collagen-rich microenvironments can successfully accumulate into multiple solid tumors, control tumor growth, and increase overall survival. SEQUENCES

[0165] SEQ ID NO: 1. IL5R-alpha amino acid sequence

[0166] MIIVAHVLLILLGATEILQADLLPDEKISLLPPVNFTIKVTGLAQVLLQWKPNPDQEQRNV NLEYQVKINAPKEDD YETRITESKC VTILHKGF S AS VRTILQNDHSLLAS S WAS AELHAP PGSPGTSIVNLTCTTNTTEDNYSRLRSYQVSLHCTWLVGTDAPEDTQYFLYYRYGSWTE ECQEYSKDTLGRNIACWFPRTFILSKGRDWLAVLVNGSSKHSAIRPFDQLFALHAIDQIN PPLNVTAEIEGTRLSIQWEKPVSAFPIHCFDYEVKIHNTRNGYLQIEKLMTNAFISIIDDLS KYDVQVRAAVSSMCREAGLWSEWSQPIYVGNDEHKPLREWFVIVIMATICFILLILSLIC KICHLWIKLFPPIPAPKSNIKDLFVTTNYEKAGSSETEIEVICYIEKPGVETLEDSVF SEQ ID NO: 2. IL5R-alpha nucleic acid sequence atgatcatcgtggcccacgtgctgctgattctgctgggagccacagagatcctgcaggccgatctgctgcccgacgagaagatttctctgct gcctcctgtgaacttcaccatcaaagtgaccggactggctcaggtgctgctgcagtggaagcccaatcctgaccaagagcagcggaacgt gaacctggaataccaagtgaagatcaacgcccctaaagaggacgactacgagacacggatcaccgagtctaagtgcgtgaccatcctgc acaagggcttctctgccagcgtgcgcacaatcctgcagaacgatcacagcctgctggcctctagctgggcctctgcagaattgcatgcccc acctggaagccctggcaccagcatcgtgaatctgacctgcaccaccaacaccaccgaggacaactacagcagactgcggagctaccag gtgtccctgcactgtacatggctcgtgggaacagacgcccctgaggatacccagtacttcctgtactacagatacggcagctggaccgagg aatgccaagagtacagcaaggacaccctgggcagaaatatcgcctgctggttccccagaaccttcatcctgagcaagggcagagattggc tggccgtgctggtcaatggcagctctaagcacagcgccatcagacccttcgaccagctgtttgccctgcacgccatcgaccagatcaaccc tccactgaatgtgaccgccgagatcgagggcaccagactgtctatccagtgggagaagcccgtgtccgcctttcctatccactgcttcgact acgaagtcaagatccacaacacccggaacggctacctgcagatcgagaagctgatgaccaacgccttcatcagcatcatcgacgacctgt ctaagtacgacgtgcaagtgcgggctgccgtgtccagcatgtgtagagaagctggcctttggagcgagtggtcccagcctatctacgtggg caacgacgagcacaagcccctgcgagagtggttcgtgatcgtgatcatggccaccatctgctttatcctgctgatcctgagcctgatctgcaa gatctgccacctgtggatcaagctgttccctcctattcctgctcctaagagcaacatcaaggacctgttcgtgaccaccaactacgagaaggc cggcagcagcgagacagagatcgaagtgatctgctacatcgagaaacccggcgtggaaaccctggaagatagcgtgttc SEQ ID NO: 3. IL5R-beta amino acid sequence VLAQGLLSMALLALCWERSLAGAEETIPLQTLRCYNDYTSHITCRWADTQDAQRLVNV TLIRRVNEDLLEPVSCDLSDDMPWSACPHPRCVPRRCVIPCQSFVVTDVDYFSFQPDRPL GTRLTVTLTQHVQPPEPRDLQISTDQDHFLLTWSVALGSPQSHWLSPGDLEFEWYKRL QD S WED AAILL SNT SQ ATLGPEHLMP S S T YVARVRTRL APGSRL S GRP SKW SPE VC WD S QPGDEAQPQNLECFFDGAA VLSC S WEVRKEVAS S VSFGLF YKP SPD AGEEEC SP VLREG LGSLHTRHHCQIPVPDPATHGQYIVSVQPRRAEKHIKSSVNIQMAPPSLNVTKDGDSYSL RWETMKMRYEHIDHTFEIQYRKDTATWKDSKTETLQNAHSMALPALEPSTRYWARVR VRTSRTGYNGIWSEWSEARSWDTESVLPMWVLALIVIFLTIAVLLALRFCGIYGYRLRRK WEEKIPNPSKSHLFQNGSAELWPPGSMSAFTSGSPPHQGPWGSRFPELEGVFPVGFGDSE VSPLTIEDPKHVCDPPSGPDTTPAASDLPTEQPPSPQPGPPAASHTPEKQASSFDFNGPYL GPPHSRSLPDILGQPEPPQEGGSQKSPPPGSLEYLCLPAGGQVQLVPLAQAMGPGQAVEV ERRPSQGAAGSPSLESGGGPAPPALGPRVGGQDQKDSPVAIPMSSGDTEDPGVASGYVS SADLVFTPNSGASSVSLVPSLGLPSDQTPSLCPGLASGPPGAPGPVKSGFEGYVELPPIEG RSPRSPRNNPVPPEAKSPVLNPGERPADVSPTSPQPEGLLVLQQVGDYCFLPGLGPGPLSL RSKPSSPGPGPEIKNLDQAFQVKKPPGQAVPQVPVIQLFKALKQQDYLSLPPWEVNKPG EVC

[0167] SEQ ID NO: 4. IL5R-beta nucleic acid sequence gttctggctcagggcctgctgtctatggctctgctggctctgtgttgggagagaagtctggctggcgccgaggaaacaatccctctgcagac cctgcggtgctacaacgactacaccagccacatcacctgtagatgggccgacacacaggacgcccagagactggtcaatgtgaccctgat cagaagagtgaacgaggacctgctggaacccgtgtcctgtgacctgagcgacgatatgccttggagcgcctgtcctcatcctagatgtgtg cctcggagatgcgtgatcccctgccagagctttgtggtcaccgacgtggactacttcagcttccagcctgacagacccctgggcaccagac tgacagtgacactgacacagcacgtgcagcctccagagcctagggacctgcagatctctaccgaccaggaccacttcctgctgacttgga gtgtggccctgggctctcctcagtctcattggcttagccctggcgacctggaattcgaggtggtgtacaagagactgcaggacagctggga agatgccgccatcctgctgagcaataccagccaggctacactgggacccgagcacctgatgcctagctctacctatgtggccagagtgcg gacaagactggcccctggatctagactgagcggcagaccttctaagtggtcccctgaagtgtgctgggatagccagcctggggatgaagc ccagcctcagaacctggaatgcttcttcgatggcgccgctgtgctgagctgttcttgggaagtgcggaaagaggtggccagcagcgttagc ttcggcctgttctacaagccctctcctgacgccggcgaagaggaatgttctcccgtgctgagagaaggcctgggcagcctgcacacaaga caccactgtcagatccccgtgcctgatcctgccacacacggccagtatatcgtgtccgtgcagccaagaagggccgagaagcacatcaag agcagcgtgaacatccagatggcccctccaagcctgaacgtgaccaaggacggcgacagctacagcctgagatgggagacaatgaaga tgcgctacgagcacatcgaccacaccttcgagatccagtaccggaaggataccgccacctggaaggacagcaagaccgagacactgca gaacgcccactctatggccctgccagctctggaacctagcaccagatactgggccagagtccgcgtgcggaccagcagaacaggctaca atggcatttggagcgagtggagcgaagccagaagctgggataccgagagcgtgctgcctatgtgggtgctcgccctgatcgtgatcttcct gacaatcgccgtgctgctggcactgagatctgcggcatctacggctacagactgcggcggaagtgggaagagaagatccccaatcctag caagagccacctgttccagaatggcagcgccgaactttggcctcctggcagcatgagcgcctttacatctggcagccctccacatcaaggc ccctggggctctagattccctgaactggaaggcgtgttccccgtcggctttggcgatagcgaagtgtcccctctgaccatcgaggaccctaa gcacgtgtgcgatcctccaagcggccctgatacaacaccagccgcctctgatctgcctaccgagcaacctcctagtcctcagcctggacct cctgccgcttctcacacacctgagaaacaggccagctccttcgacttcaacggcccctatctgggcccacctcactctagaagcctgcctga tatcctgggacagcccgagcctcctcaagaaggcggatctcagaagtctcctccacctggcagcctggaatacctgtgtttgcctgctggtg gccaggtgcagctggttcctcttgctcaagctatgggacctggacaggccgtggaagtggaaagaaggccatctcaaggcgctgccggat ctccttctctggaatctggcggaggaccagctcctcctgctctgggaccaagagttggaggccaggaccagaaagacagccccgtggcca ttcctatgagcagcggcgatactgaggatcctggcgtggcctctggctatgtgtctagcgccgacctggtgttcacccctaattctggcgcct cctccgtgtctctggtgccatctcttggactgcccagcgatcagaccccttctctgtgtcctggactggcttctggaccacctggtgctccagg acctgtgaagtctggattcgagggctacgtggaactgcctcctatcgagggcagaagccctagaagccccagaaacaaccctgtgcctcct gaggccaagtctcctgtgctgaaccctggcgaaagacccgccgatgtgtctccaacatctcctcagccagagggactgctggttctgcagc aagtgggcgactactgcttcctgccaggactggtcctggaccactgagcctgagaagcaagcctctagccctggacctgggcctgagat caagaatctggatcaggccttccaagtgaagaagcctccaggccaggctgtgcctcaggtgccagtgatccagctgttcaaggccctgaa gcagcaggactacctgagcctgcctccatgggaagtgaacaagccaggcgaagtgtgc

[0168] SEQ ID NO: 5. P2A amino acid sequence

[0169] GSGATNFSLLKQAGDVEENPGP

[0170] SEQ ID NO: 6. IL5R-alpha-P2A-IL5R-beta amino acid sequence

[0171] MIIVAHVLLILLGATEILQADLLPDEKISLLPPVNFTIKVTGLAQVLLQWKPNPDQEQRNV NLEYQVKINAPKEDD YETRITESKC VTILHKGF S AS VRTILQNDHSLLAS SWAS AELHAP PGSPGTSIVNLTCTTNTTEDNYSRLRSYQVSLHCTWLVGTDAPEDTQYFLYYRYGSWTE ECQEYSKDTLGRNIACWFPRTFILSKGRDWLAVLVNGSSKHSAIRPFDQLFALHAIDQIN PPLNVTAEIEGTRLSIQWEKPVSAFPIHCFDYEVKIHNTRNGYLQIEKLMTNAFISIIDDLS KYDVQVRAAVSSMCREAGLWSEWSQPIYVGNDEHKPLREWFVIVIMATICFILLILSLIC KICHLWIKLFPPIPAPKSNIKDLFVTTNYEKAGSSETEIEVICYIEKPGVETLEDSVFGSGAT NF SLLKQ AGD VEENPGPVL AQGLL SMALLALC WERSLAGAEETIPLQTLRC YND YT SHI TCRWADTQDAQRLVNVTLIRRVNEDLLEPVSCDLSDDMPWSACPHPRCVPRRCVIPCQS FVVTDVDYFSFQPDRPLGTRLTVTLTQHVQPPEPRDLQISTDQDHFLLTWSVALGSPQSH WLSPGDLEFEVVYKRLQDSWEDAAILLSNTSQATLGPEHLMPSSTYVARVRTRLAPGSR LSGRPSKWSPEVCWDSQPGDEAQPQNLECFFDGAAVLSCSWEVRKEVASSVSFGLFYK PSPDAGEEECSPVLREGLGSLHTRHHCQIPVPDPATHGQYIVSVQPRRAEKHIKSSVNIQ MAPP SLNVTKDGD S YSLRWETMKMRYEHIDHTFEIQ YRKDT ATWKD SKTETLQNAHS MALPALEPSTRYWARVRVRTSRTGYNGIWSEWSEARSWDTESVLPMWVLALIVIFLTIA VLLALRFCGIYGYRLRRKWEEKIPNPSKSHLFQNGSAELWPPGSMSAFTSGSPPHQGPW GSRFPELEGVFPVGFGDSEVSPLTIEDPKHVCDPPSGPDTTPAASDLPTEQPPSPQPGPPAA SHTPEKQASSFDFNGPYLGPPHSRSLPDILGQPEPPQEGGSQKSPPPGSLEYLCLPAGGQV QLVPLAQAMGPGQAVEVERRPSQGAAGSPSLESGGGPAPPALGPRVGGQDQKDSPVAIP MSSGDTEDPGVASGYVSSADLVFTPNSGASSVSLVPSLGLPSDQTPSLCPGLASGPPGAP GPVKSGFEGYVELPPIEGRSPRSPRNNPVPPEAKSPVLNPGERPADVSPTSPQPEGLLVLQ QVGDYCFLPGLGPGPLSLRSKPSSPGPGPEIKNLDQAFQVKKPPGQAVPQVPVIQLFKAL KQQDYLSLPPWEVNKPGEVC

[0172] SEQ ID NO: 7. P2A nucleic acid sequence ggatccggagccacgaacttctctctgttaaagcaagcaggcgacgtggaagaaaaccccggtccc

[0173] SEQ ID NO: 8. IL5R-alpha-P2A-IL5R-beta nucleic acid sequence atgatcatcgtggcccacgtgctgctgattctgctgggagccacagagatcctgcaggccgatctgctgcccgacgagaagatttctctgct gcctcctgtgaacttcaccatcaaagtgaccggactggctcaggtgctgctgcagtggaagcccaatcctgaccaagagcagcggaacgt gaacctggaataccaagtgaagatcaacgcccctaaagaggacgactacgagacacggatcaccgagtctaagtgcgtgaccatcctgc acaagggcttctctgccagcgtgcgcacaatcctgcagaacgatcacagcctgctggcctctagctgggcctctgcagaattgcatgcccc acctggaagccctggcaccagcatcgtgaatctgacctgcaccaccaacaccaccgaggacaactacagcagactgcggagctaccag gtgtccctgcactgtacatggctcgtgggaacagacgcccctgaggatacccagtacttcctgtactacagatacggcagctggaccgagg aatgccaagagtacagcaaggacaccctgggcagaaatatcgcctgctggttccccagaaccttcatcctgagcaagggcagagattggc tggccgtgctggtcaatggcagctctaagcacagcgccatcagacccttcgaccagctgtttgccctgcacgccatcgaccagatcaaccc tccactgaatgtgaccgccgagatcgagggcaccagactgtctatccagtgggagaagcccgtgtccgcctttcctatccactgcttcgact acgaagtcaagatccacaacacccggaacggctacctgcagatcgagaagctgatgaccaacgccttcatcagcatcatcgacgacctgt ctaagtacgacgtgcaagtgcgggctgccgtgtccagcatgtgtagagaagctggcctttggagcgagtggtcccagcctatctacgtggg caacgacgagcacaagcccctgcgagagtggttcgtgatcgtgatcatggccaccatctgctttatcctgctgatcctgagcctgatctgcaa gatctgccacctgtggatcaagctgttccctcctattcctgctcctaagagcaacatcaaggacctgttcgtgaccaccaactacgagaaggc cggcagcagcgagacagagatcgaagtgatctgctacatcgagaaacccggcgtggaaaccctggaagatagcgtgttcggatccgga gccacgaacttctctctgttaaagcaagcaggcgacgtggaagaaaaccccggtcccgttctggctcagggcctgctgtctatggctctgct ggctctgtgttgggagagaagtctggctggcgccgaggaaacaatccctctgcagaccctgcggtgctacaacgactacaccagccacat cacctgtagatgggccgacacacaggacgcccagagactggtcaatgtgaccctgatcagaagagtgaacgaggacctgctggaaccc gtgtcctgtgacctgagcgacgatatgccttggagcgcctgtcctcatcctagatgtgtgcctcggagatgcgtgatcccctgccagagcttt gtggtcaccgacgtggactacttcagctccagcctgacagacccctgggcaccagactgacagtgacactgacacagcacgtgcagcct ccagagcctagggacctgcagatctctaccgaccaggaccacttcctgctgacttggagtgtggccctgggctctcctcagtctcattggctt agccctggcgacctggaattcgaggtggtgtacaagagactgcaggacagctgggaagatgccgccatcctgctgagcaataccagcca ggctacactgggacccgagcacctgatgcctagctctacctatgtggccagagtgcggacaagactggcccctggatctagactgagcgg cagaccttctaagtggtcccctgaagtgtgctgggatagccagcctggggatgaagcccagcctcagaacctggaatgcttcttcgatggc gccgctgtgctgagctgttcttgggaagtgcggaaagaggtggccagcagcgttagcttcggcctgttctacaagccctctcctgacgccg gcgaagaggaatgttctcccgtgctgagagaaggcctgggcagcctgcacacaagacaccactgtcagatccccgtgcctgatcctgcca cacacggccagtatatcgtgtccgtgcagccaagaagggccgagaagcacatcaagagcagcgtgaacatccagatggcccctccaag cctgaacgtgaccaaggacggcgacagctacagcctgagatgggagacaatgaagatgcgctacgagcacatcgaccacaccttcgag atccagtaccggaaggataccgccacctggaaggacagcaagaccgagacactgcagaacgcccactctatggccctgccagctctgga acctagcaccagatactgggccagagtccgcgtgcggaccagcagaacaggctacaatggcatttggagcgagtggagcgaagccaga agctgggataccgagagcgtgctgcctatgtgggtgctcgccctgatcgtgatcttcctgacaatcgccgtgctgctggcactgagattctgc ggcatctacggctacagactgcggcggaagtgggaagagaagatccccaatcctagcaagagccacctgttccagaatggcagcgccg aactttggcctcctggcagcatgagcgcctttacatctggcagccctccacatcaaggcccctggggctctagattccctgaactggaaggc gtgttccccgtcggctttggcgatagcgaagtgtcccctctgaccatcgaggaccctaagcacgtgtgcgatcctccaagcggccctgatac aacaccagccgcctctgatctgcctaccgagcaacctcctagtcctcagcctggacctcctgccgcttctcacacacctgagaaacaggcc agctccttcgacttcaacggcccctatctgggcccacctcactctagaagcctgcctgatatcctgggacagcccgagcctcctcaagaagg cggatctcagaagtctcctccacctggcagcctggaatacctgtgtttgcctgctggtggccaggtgcagctggttcctcttgctcaagctatg ggacctggacaggccgtggaagtggaaagaaggccatctcaaggcgctgccggatctccttctctggaatctggcggaggaccagctcct cctgctctgggaccaagagttggaggccaggaccagaaagacagccccgtggccattcctatgagcagcggcgatactgaggatcctgg cgtggcctctggctatgtgtctagcgccgacctggtgttcacccctaattctggcgcctcctccgtgtctctggtgccatctcttggactgccca gcgatcagaccccttctctgtgtcctggactggcttctggaccacctggtgctccaggacctgtgaagtctggattcgagggctacgtggaa ctgcctcctatcgagggcagaagccctagaagccccagaaacaaccctgtgcctcctgaggccaagtctcctgtgctgaaccctggcgaa agacccgccgatgtgtctccaacatctcctcagccagagggactgctggttctgcagcaagtgggcgactactgcttcctgccaggacttgg tcctggaccactgagcctgagaagcaagccttctagccctggacctgggcctgagatcaagaatctggatcaggccttccaagtgaagaag cctccaggccaggctgtgcctcaggtgccagtgatccagctgttcaaggccctgaagcagcaggactacctgagcctgcctccatgggaa gtgaacaagccaggcgaagtgtgctaa

[0174] SEQ ID NO: 9. IL5 amino acid sequence MRMLLHLSLLALGAAYVYAIPTEIPTSALVKETLALLSTHRTLLIANETLRIPVPVHKNH

[0175] QLCTEEIFQGIGTLESQTVQGGTVERLFKNLSLIKKYIDGQKKKCGEERRRVNQFLDYLQ EFLGVMNTEWIIES

[0176] SEQ ID NO: 10. IL5R-alpha binding domain and hinge domain

[0177] MIIVAHVLLILLGATEILQADLLPDEKISLLPPVNFTIKVTGLAQVLLQWKPNPDQEQRNV NLEYQ VKINAPKEDD YETRITE SKC VTILHKGF S AS VRTILQNDHSLLAS S WAS AELHAP PGSPGTSIVNLTCTTNTTEDNYSRLRSYQVSLHCTWLVGTDAPEDTQYFLYYRYGSWTE ECQEYSKDTLGRNIACWFPRTFILSKGRDWLAVLVNGSSKHSAIRPFDQLFALHAIDQIN PPLNVTAEIEGTRLSIQWEKPVSAFPIHCFDYEVKIHNTRNGYLQIEKLMTNAFISIIDDLS KYDVQVRAAVSSMCREAGLWSEWSQPIYVGNDEHKPLREW

[0178] SEQ ID NO: 11. IL5R-alpha transmembrane domain

[0179] FVIVIMATICFILLILSLIC

[0180] SEQ ID NO: 12. IL5R-alpha signaling domain

[0181] KICHLWIKLFPPIPAPKSNIKDLFVTTNYEKAGSSETEIEVICYIEKPGVETLEDSVF

[0182] SEQ ID NO: 13. IL5R-beta binding domain and hinge domain

[0183] VLAQGLLSMALLALCWERSLAGAEETIPLQTLRCYNDYTSHITCRWADTQDAQRLVNV TLIRRVNEDLLEPVSCDLSDDMPWSACPHPRCVPRRCVIPCQSFVVTDVDYFSFQPDRPL GTRLTVTLTQHVQPPEPRDLQISTDQDHFLLTWSVALGSPQSHWLSPGDLEFEVVYKRL QD S WED AAILL SNT SQ ATLGPEHLMP S S T YVARVRTRL APGSRL S GRP SKW SPE VC WD S QPGDEAQPQNLECFFDGAAVLSCSWEVRKEVASSVSFGLFYKPSPDAGEEECSPVLREG LGSLHTRHHCQIPVPDPATHGQYIVSVQPRRAEKHIKSSVNIQMAPPSLNVTKDGDSYSL RWETMKMRYEHIDHTFEIQYRKDTATWKDSKTETLQNAHSMALPALEPSTRYWARVR VRTSRTGYNGIWSEWSEARSWDTESVLPMW

[0184] SEQ ID NO: 14. IL5R-beta transmembrane domain

[0185] VLALIVIFLTIAVLLAL

[0186] SEQ ID NO: 15. IL5R-beta signaling domain

[0187] RFCGIYGYRLRRKWEEKIPNPSKSHLFQNGSAELWPPGSMSAFTSGSPPHQGPWGSRFPE LEGVFPVGFGDSEVSPLTIEDPKHVCDPPSGPDTTPAASDLPTEQPPSPQPGPPAASHTPE KQASSFDFNGPYLGPPHSRSLPDILGQPEPPQEGGSQKSPPPGSLEYLCLPAGGQVQLVPL AQAMGPGQAVEVERRPSQGAAGSPSLESGGGPAPPALGPRVGGQDQKDSPVAIPMSSG DTEDPGVASGYVSSADLVFTPNSGASSVSLVPSLGLPSDQTPSLCPGLASGPPGAPGPVK SGFEGYVELPPIEGRSPRSPRNNPVPPEAKSPVLNPGERPADVSPTSPQPEGLLVLQQVGD YCFLPGLGPGPLSLRSKPSSPGPGPEIKNLDQAFQVKKPPGQAVPQVPVIQLFKALKQQD YLSLPPWEVNKPGEVC

[0188] SEQ ID NO: 16. VH chain of IL5 1995 scfv binding domain

[0189] ESGGGLVQPSQTLSLTCTVSGLSLTSNSVNWIRQPPGKGLEWMGLIWSNGDTDYNSAIK SRLSISRDTSKSQVFLKMNSLQSEDTAMYFCAREYYGYFDYWGQGVMVTVSS

[0190] SEQ ID NO: 17. VL chain of IL5 1995 scfv binding domain AVPTQLLGLLLLWITDAICDIQMTQSPASLSASLGETISIECLASEGISSYLAWYQQKPGK SPQLLIYGANSLQTGVPSRFSGSGSATQYSLKISSMQPEDEGDYFCQQSYKFPNTFGAGT KLELKR

[0191] SEQ ID NO: 18. TNF-alpha amino acid sequence

[0192] MSTESMIRDVELAEEALPKKTGGPQGSRRCLFLSLFSFLIVAGATTLFCLLHFGVIGPQRE

[0193] EFPRDLSLISPLAQAVRSSSRTPSDKPVAHVVANPQAEGQLQWLNRRANALLANGVELR DNQLVVPSEGLYLIYSQVLFKGQGCPSTHVLLTHTISRIAVSYQTKVNLLSAIKSPCQRET PEGAEAKPWYEPIYLGGVFQLEKGDRLSAEINRPDYLDFAESGQVYFGIIAL

[0194] SEQ ID NO: 19. TNFR1 binding domain and hinge domain

[0195] MGLSTVPDLLLPLVLLELLVGIYPSGVIGLVPHLGDREKRDSVCPQGKYIHPQNNSICCT

[0196] KCHKGTYLYNDCPGPGQDTDCRECESGSFTASENHLRHCLSCSKCRKEMGQVEISSCTV DRDTVCGCRKNQYRHYWSENLFQCFNCSLCLNGTVHLSCQEKQNTVCTCHAGFFLREN ECVSCSNCKKSLECTKLCLPQIENVKGTEDSGTT

[0197] SEQ ID NO: 20. TNFR1 transmembrane domain

[0198] VLLPLVIFFGLCLLSLLFIGL

[0199] SEQ ID NO: 21. TNFR1 signaling domain

[0200] MYRYQRWKSKLYSIVCGKSTPEKEGELEGTTTKPLAPNPSFSPTPGFTPTLGFSPVPSSTF T S S ST YTPGDCPNF AAPRRE VAPP YQGADPIL ATAL ASDPIPNPLQKWED S AHKPQ SLDT DDPATLYAVVENVPPLRWKEFVRRLGLSDHEIDRLELQNGRCLREAQYSMLATWRRRT PRREATLELLGRVLRDMDLLGCLEDIEEALCGPAALPPAPSLLR

[0201] SEQ ID NO: 22. TNFR2 binding domain and hinge domain

[0202] MAPVAVWAALAVGLELWAAAHALPAQVAFTPYAPEPGSTCRLREYYDQTAQMCCSK

[0203] CSPGQHAKVFCTKTSDTVCDSCEDSTYTQLWNWVPECLSCGSRCSSDQVETQACTREQ NRICTCRPGWYCALSKQEGCRLCAPLRKCRPGFGVARPGTETSDVVCKPCAPGTFSNTT

[0204] SSTDICRPHQICNVVAIPGNASMDAVCTSTSPTRSMAPGAVHLPQPVSTRSQHTQPTPEPS TAPSTSFLLPMGPSPPAEGSTGD

[0205] SEQ ID NO: 23. TNFR2 transmembrane domain

[0206] FALPVGLIVGVTALGLLIIGVVNCVIMTQV

[0207] SEQ ID NO: 24. TNFR2 signaling domain KKKPLCLQRE AK VPHLP ADK ARGTQGPEQQHLLIT AP S S S S S SLES S A S ALDRR APTRNQ PQ APGVE ASGAGEARASTGS SD S SPGGHGTQ VNVTCIVNVC S S SDHS SQC S SQ AS STMG DTD S SP SESPKDEQ VPF SKEEC AFRSQLETPETLLGSTEEKPLPLGVPD AGMKP S SEQ ID NO: 25. VH chain of TNF-alpha D2E7 scfv binding domain EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHI DYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS

[0208] SEQ ID NO: 197. VH chain of TNF-alpha TSA2 scfv binding domain QVQLQESGPGLVKPSETLSLTCTVSINYTGEPTYSQSVSNVVWYWIRQPPGKGLEWIGY TYYPTVNENRSQSVSNVFRVTISVDTSKNQFSLKLSSVTAADTAVYYCARSSYPGYSQPT RSYPSSWGQGTL VTVSS

[0209] SEQ ID NO: 26. VL chain of TNF-alpha D2E7 scfv binding domain2

[0210] DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPS RFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKAAAHHHHHHGEQ KLISEEDL

[0211] SEQ ID NO: 198. VL chain of TNF-alpha TSA2 scfv binding domain3

[0212] QSVLTQPPSVSGAPGQRVTISCSGSSSNIGSNYVSWYQQLPGTAPKLLIYDNNQRPSGVP DRFSGSKSGTSASLAITGLQSEDEADYYCYINTGYDGLYYNSMDFGGGTKLTVLG SEQ ID NO: 27. IFN-gamma amino acid sequence MKYTSYILAFQLCIVLGSLGCYCQDPYVKEAENLKKYFNAGHSDVADNGTLFLGILKN WKEESDRKIMQSQIVSFYFKLFKNFKDDQSIQKSVETIKEDMNVKFFNSNKKKRDDFEK LTNYSVTDLNVQRKAIHELIQVMAELSPAAKTGKRKRSQMLFRGRRASQ SEQ ID NO: 28. IFN-gammaRl binding domain and hinge domain

[0213] MALLFLLPLVMQGVSRAEMGTADLGPSSVPTPTNVTIESYNMNPIVYWEYQIMPQVPVF TVEVKNYGVKNSEWIDACINISHHYCNISDHVGDPSNSLWVRVKARVGQKESAYAKSE EFAVCRDGKIGPPKLDIRKEEKQIMIDIFHPSVFVNGDEQEVDYDPETTCYIRVYNVYVR MNGSEIQYKILTQKEDDCDEIQCQLAIPVSSLNSQYCVSAEGVLHVWGVTTEKSKEVCIT IFNSSIKG

[0214] SEQ ID NO: 29. IFN-gammaRl transmembrane domain SLWIP VV AALLLFL VL SLVFI

[0215] SEQ ID NO: 30. IFN-gammaRl signaling domain CFYIKKINPLKEKSIILPKSLISVVRSATLETKPESKYVSLITSYQPFSLEKEVVCEEPLSPAT VPGMHTEDNPGKVEHTEELSSITEVVTTEENIPDVVPGSHLTPIERESSSPLSSNQSEPGSI ALNSYHSRNCSESDHSRNGFDTDSSCLESHSSLSDSEFPPNNKGEIKTEGQELITVIKAPTS FGYDKPHVLVDLLVDDSGKESLIGYRPTEDSKEFS

[0216] SEQ ID NO: 31. IFN-gammaR2 binding domain and hinge domain

[0217] RPTLLWSLLLLLGVFAAAAAAPPDPLSQLPAPQHPKIRLYNAEQVLSWEPVALSNSTRPV

[0218] VYQVQFKYTDSKWFTADIMSIGVNCTQITATECDFTAASPSAGFPMDFNVTLRLRAELG ALHSAWVTMPWFQHYRNVTVGPPENIEVTPGEGSLIIRFSSPFDIADTSTAFFCYYVHYW EKGGIQQVKGPFRSNSISLDNLKPSRVYCLQVQAQLLWNKSNIFRVGHLSNISCYETMA DASTELQQ

[0219] SEQ ID NO: 32. IFN-gammaR2 transmembrane domain

[0220] VILISVGTFSLLSVLAGACFF

[0221] SEQ ID NO: 33. IFN-gammaR2 signaling domain

[0222] LVLKYRGLIKYWFHTPPSIPLQIEEYLKDPTQPILEALDKDSSPKDDVWDSVSIISFPEKEQ EDVLQTL

[0223] SEQ ID NO: 34. VH chain of IFN-gamma NI-0501 scfv binding domain

[0224] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTY

[0225] YADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDGSSGWYVPHWFDPWGQG TLVTVSS

[0226] SEQ ID NO: 35. VL chain of IFN-gamma NI-0501 scfv binding domain

[0227] NFMLTQPHSVSESPGKTVTISCTRSSGSIASNYVQWYQQRPGSSPTTVIYEDNQRPSGVP

[0228] DRF SGSID S S SNS ASLTISGLKTEDEAD YYCQ S YDGSNRWMFGGGTKLT VL

[0229] SEQ ID NO: 36. IL8 amino acid sequence

[0230] MTSKLAVALLAAFLISAALCEGAVLPRSAKELRCQCIKTYSKPFHPKFIKELRVIESGPHC

[0231] ANTEIIVKLSDGRELCLDPKENWVQRVVEKFLKRAENS

[0232] SEQ ID NO: 37. IL8RA binding domain and hinge domain 1

[0233] MSNITDPQMWDFDDLNFTGMPPADEDYSPCMLETETLNK

[0234] SEQ ID NO: 38. IL8RA binding domain and hinge domain 2

[0235] ASKVNGWIFGTFLCK

[0236] SEQ ID NO: 39. IL8RA binding domain and hinge domain 3

[0237] RQ A YHPNNS SP VC YEVLGNDTAKWR SEQ ID NO: 40. IL8RA binding domain and hinge domain 4

[0238] DTLMRTQVIQESCERRNNIGR

[0239] SEQ ID NO: 41. IL8RA transmembrane domain 1

[0240] YVVIIAYAL VFLL SLLGNSL VML VIL Y

[0241] SEQ ID NO: 42. IL8RA transmembrane domain 2

[0242] VYLLNLALADLLFALTLPIWA

[0243] SEQ ID NO: 43. IL8RA transmembrane domain 3

[0244] VVSLLKEVNFYSGILLLACISV

[0245] SEQ ID NO: 44. IL8RA transmembrane domain 4

[0246] FVCLGCWGLSMNLSLPFFLF

[0247] SEQ ID NO: 45. IL8RA transmembrane domain 5

[0248] MVLRFLPHTFGFIVPLFVMLF

[0249] SEQ ID NO: 46. IL8RA transmembrane domain 6

[0250] VIFAVVLIFLLCWLPYNLVLLA

[0251] SEQ ID NO: 47. IL8RA transmembrane domain 7

[0252] ALDATEILGFLHSCLNPIIYAFI

[0253] SEQ ID NO: 48. IL8RA signaling domain 1

[0254] SRVGRSVTD

[0255] SEQ ID NO: 49. IL8RA signaling domain 2

[0256] DRYLAIVHATRTLTQKRHLVK

[0257] SEQ ID NO: 50. IL8RA signaling domain 3

[0258] CYGFTLRTLFKAHMGQKHRAMR

[0259] SEQ ID NO: 51. IL8RA signaling domain 4

[0260] GQNFRHGFLKILAMHGLVSKEFLARHRVTS YTS S S VNVS SNL

[0261] SEQ ID NO: 52. IL8RA signaling domain 5 (fusion of all IL8RA signaling domains)

[0262] SRVGRSVTDDRYLAIVHATRTLTQKRHLVKCYGFTLRTLFKAHMGQKHRAMRGQNFR

[0263] HGFLKILAMHGLVSKEFLARHRVTS YTS S S VNVSSNL

[0264] SEQ ID NO: 53. IL8RB binding domain and hinge domain 1

[0265] MEDFNMESDSFEDFWKGEDLSNYSYSSTLPPFLLDAAPCEPESLEINK

[0266] SEQ ID NO: 54. IL8RB binding domain and hinge domain 2

[0267] ASKVNGWIFGTFLCK SEQ ID NO: 55. IL8RB binding domain and hinge domain 3

[0268] RRTVYS SNVSPAC YEDMGNNTANWR

[0269] SEQ ID NO: 56. IL8RB binding domain and hinge domain 4

[0270] DTLMRTQVIQETCERRNHIDR

[0271] SEQ ID NO: 57. IL8RB transmembrane domain 1

[0272] YFVVIIYALVFLLSLLGNSLVMLVILY

[0273] SEQ ID NO: 58. IL8RB transmembrane domain 2

[0274] VYLLNLALADLLFALTLPIWA

[0275] SEQ ID NO: 59. IL8RB transmembrane domain 3

[0276] VVSLLKEVNFYSGILLLACISV

[0277] SEQ ID NO: 60. IL8RB transmembrane domain 4

[0278] FICLSIWGLSLLLALPVLLF

[0279] SEQ ID NO: 61. IL8RB transmembrane domain 5

[0280] MLLRILPQSFGFIVPLLIMLFCY

[0281] SEQ ID NO: 62. IL8RB transmembrane domain 6

[0282] VIFAVVLIFLLCWLPYNLVLLA

[0283] SEQ ID NO: 63. IL8RB transmembrane domain 7

[0284] ALDATEILGILHSCLNPLIYA

[0285] SEQ ID NO: 64. IL8RB signaling domain 1

[0286] SRVGRSVTD

[0287] SEQ ID NO: 65. IL8RB signaling domain 2

[0288] DRYLAIVHATRTLTQKRYLVK

[0289] SEQ ID NO: 66. IL8RB signaling domain 3

[0290] GFTLRTLFKAHMGQKHRAMR

[0291] SEQ ID NO: 67. IL8RB signaling domain 4

[0292] FIGQKFRHGLLKIL AIHGLISKD SLPKD SRPSFVGSSS GHT S TTL

[0293] SEQ ID NO: 68. IL8RB signaling domain 5 (fusion of all IL8RB signaling domains)

[0294] SRVGRSVTDDRYLAIVHATRTLTQKRYLVKGFTLRTLFKAHMGQKHRAMRFIGQKFRH

[0295] GLLKILAIHGLISKDSLPKDSRPSFVGSSSGHTSTTL

[0296] SEQ ID NO: 69. VH chain of IL8 6G4.2.5 scfv binding domain EIQLQQSGPELMKPGASVKISCKASGYSFSSHYMHWVKQSHGKSLEWIGYIDPSNGETT

[0297] YNQKFKGKATLTVDTSSSTANVHLSSLTSDDSAVYFCARGDYRYNGDWFFDVWGAGT TVTVS S

[0298] SEQ ID NO: 70. VL chain of IL8 6G4.2.5 scfv binding domain

[0299] DIVMTQTPLSLPVSLGDQASISCRSSQSLVHGIGNTYLHWYLQKPGQSPKLLIYKVSNRF

[0300] SGVPDRFSGSGSGTDFTLRISRVEAEDLGLYFCSQSTHVPLTFGAGTKLELKRA

[0301] SEQ ID NO: 71. VH chain of IL8 6G4.2.5V11 scfv binding domain

[0302] EVQLVESGGGLVQPGGSLRLSCAASGYSFSSHYMHWVRQAPGKGLEWVGYIDPSNGET

[0303] TYNQKFKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGDYRYNGDWFFDVWGQ GT

[0304] SEQ ID NO: 72. VL chain of IL8 6G4.2.5V11 scfv binding domain

[0305] DIQMTQSPSSLSASVGDRVTITCRSSQSLVHGIGNTYLHWYQQKPGKAPKLLIYKVSNRF

[0306] SGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCSQSTHVPLTFGQGTKVEIKR

[0307] Hinge

[0308] SEQ ID NO: 73. CD8a hinge domain

[0309] TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD

[0310] SEQ ID NO: 74. CD4 hinge domain

[0311] SKREKAVWVLNPEAGMWQCLLSDSGQVLLESNIKVLPTWSTPVQP

[0312] SEQ ID NO: 75. IgG4 hinge domain

[0313] ESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWY

[0314] VDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTIS

[0315] KAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP

[0316] VLD SDGSFFLYSRLT VDK SRWQEGNVF SC S VMHEALHNHYTQKSLSL SLGK

[0317] SEQ ID NO: 76. IgD hinge domain

[0318] RWPESPKAQASSVPTAQPQAEGSLAKATTAPATTRNTGRGGEEKKKEKEKEEQEERETK

[0319] TPECP

[0320] SEQ ID NO: 77. CD28 hinge domain

[0321] IEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP

[0322] Transmembrane

[0323] SEQ ID NO: 78. CD8a transmembrane domain

[0324] IYIWAPLAGTCGVLLLSLVITLYC SEQ ID NO: 79. CD4 transmembrane domain

[0325] MALIVLGGVAGLLLFIGLGIFF

[0326] SEQ ID NO: 80. CD28 transmembrane domain

[0327] FWVLVVVGGVLACYSLLVTVAFIIFWV

[0328] Transmembrane domain including a portion of the respective intracellular domain

[0329] SEQ ID NO: 81. CD8a transmembrane domain including a portion of the intracellular domain

[0330] I YIW APL AGTC GVLLL SL VITL YCL YC

[0331] SEQ ID NO: 82. CD4 transmembrane domain

[0332] M ALIVE GGVAGLLLFIGLGIFFCVR

[0333] SEQ ID NO: 83. CD28 transmembrane domain

[0334] FWVLVVVGGVLACYSLLVTVAFIIFWVRSK

[0335] Linker

[0336] SEQ ID NO: 84. scFvLinker

[0337] GGGGSGGGGSGGGGSGGGGS

[0338] Tags

[0339] SEQ ID NO: 85. 8X His tag

[0340] GSHHHHHHHH

[0341] SEQ ID NO: 86. V5 tag

[0342] GKPIPNPLLGLDST

[0343] SEQ ID NO: 87. HA tag

[0344] YPYDVPDYA

[0345] SEQ ID NO: 88. Myc tag

[0346] EQKLISEEDL

[0347] SEQ ID NO: 89. FLAG tag

[0348] DYKDDDDK

[0349] SEQ ID NO: 90. HQ tag

[0350] HQHQHQ

[0351] SEQ ID NO: 91. HN tag

[0352] HNHNHNHNHNHN

[0353] SEQ ID NO: 92. HAT tag

[0354] KDHI.IHNVHKF.F.HAHAHNK Signal peptide

[0355] SEQ ID NO: 93. CD8a signal peptide

[0356] MSLPVTALLLPLALLLHAARP

[0357] SEQ ID NO: 94. IgGl signal peptide

[0358] MAVPTQVLGLLLLWLTDARC

[0359] Signaling

[0360] SEQ ID NO: 95. Arp2

[0361] MDSQGRKVVVCDNGTGFVKCGYAGSNFPEHIFPALVGRPIIRSTTKVGNIEIKDLMVGD EASELRSMLEVNYPMENGIVRNWDDMKHLWDYTFGPEKLNIDTRNCKILLTEPPMNPT KNREKIVE VMFETYQF SGVYVAIQ AVLTLYAQGLLTGVVVD SGDGVTHICP VYEGF SLP HLTRRLDIAGRDITRYLIKLLLLRGYAFNHSADFETVRMIKEKLCYVGYNIEQEQKLALE TTVLVESYTLPDGRIIKVGGERFEAPEALFQPHLINVEGVGVAELLFNTIQAADIDTRSEF YKHIVLSGGSTMYPGLPSRLERELKQLYLERVLKGDVEKLSKFKIRIEDPPRRKHMVFLG GAVLADIMKDKDNFWMTRQEYQEKGVRVLEKLGVTVR

[0362] SEQ ID NO: 96. Arp3

[0363] MAGRLPACVVDCGTGYTKLGYAGNTEPQFIIPSCIAIKESAKVGDQAQRRVMKGVDDL DFFIGDEAIEKPTYATKWPIRHGIVEDWDLMERFMEQVIFKYLRAEPEDHYFLLTEPPLN TPENREYTAEIMFESFNVPGLYIAVQAVLALAASWTSRQVGERTLTGTVIDSGDGVTHVI PVAEGYVIGSCIKHIPIAGRDITYFIQQLLRDREVGIPPEQSLETAKAVKERYSYVCPDLV KEFNKYDTDGSKWIKQYTGINAISKKEFSIDVGYERFLGPEIFFHPEFANPDFTQPISEVV DEVIQNCPIDVRRPLYKNIVLSGGSTMFRDFGRRLQRDLKRTVDARLKLSEELSGGRLKP KPIDVQVITHHMQRYAVWFGGSMLASTPEFYQVCHTKKDYEEIGPSICRHNPVFGVMS SEQ ID NO: 97. ActB

[0364] MDDDIAALVVDNGSGMCKAGFAGDDAPRAVFPSIVGRPRHQGVMVGMGQKDSYVGD EAQSKRGILTLKYPIEHGIVTNWDDMEKIWHHTFYNELRVAPEEHPVLLTEAPLNPKAN REKMTQIMFETFNTPAMYVAIQAVLSLYASGRTTGIVMDSGDGVTHTVPIYEGY ALPHA ILRLDLAGRDLTDYLMKILTERGYSFTTTAEREIVRDIKEKLC YVALDFEQEMATAAS S S SLEKSYELPDGQVITIGNERFRCPEALFQPSFLGMESCGIHETTFNSIMKCDVDIRKDLYA NTVLSGGTTMYPGIADRMQKEITALAPSTMKIKIIAPPERKYSVWIGGSILASLSTFQQM WISKQEYDESGPSIVHRKCF

[0365] SEQ ID NO: 98. ActGl MEEEIAALVIDNGSGMCKAGFAGDDAPRAVFPSIVGRPRHQGVMVGMGQKDSYVGDE AQSKRGILTLKYPIEHGIVTNWDDMEKIWHHTFYNELRVAPEEHPVLLTEAPLNPKANR EKMTQIMFETFNTPAMYVAIQAVLSLYASGRTTGIVMDSGDGVTHTVPIYEGYALPHAI LRLDL AGRDLTDYLMKILTERGYSFTTTAEREIVRDIKEKLCYVALDFEQEMATAAS S S S LEKSYELPDGQVITIGNERFRCPEALFQPSFLGMESCGIHETTFNSIMKCDVDIRKDLYAN TVLSGGTTMYPGIADRMQKEITALAPSTMKIKIIAPPERKYSVWIGGSILASLSTFQQMWI SKQE YDE S GP SIVHRKCF SEQ ID NO: 99. ActG2

[0366] MCEEETTALVCDNGSGLCKAGFAGDDAPRAVFPSIVGRPRHQGVMVGMGQKDSYVGD EAQSKRGILTLKYPIEHGIITNWDDMEKIWHHSFYNELRVAPEEHPTLLTEAPLNPKANR EKMTQIMFETFNVPAMYVAIQAVLSLYASGRTTGIVLDSGDGVTHNVPIYEGY ALPHAI MRLDL AGRDLTD YLMKILTERGYSF VTT AEREIVRDIKEKLC YV ALDFENEMAT AAS S S SLEKSYELPDGQVITIGNERFRCPETLFQPSFIGMESAGIHETTYNSIMKCDIDIRKDLYAN NVLSGGTTMYPGIADRMQKEITALAPSTMKIKIIAPPERKYSVWIGGSILASLSTFQQMWI SKPEYDE AGP SIVHRKCF SEQ ID NO: 100. WASP

[0367] MSGGPMGGRPGGRGAPAVQQNIPSTLLQDHENQRLFEMLGRKCLTLATAVVQLYLALP PGAEHWTKEHCGAVCFVKDNPQKSYFIRLYGLQAGRLLWEQELYSQLVYSTPTPFFHTF AGDDCQAGLNFADEDEAQAFRALVQEKIQKRNQRQSGDRRQLPPPPTPANEERRGGLP PLPLHPGGDQGGPPVGPLSLGLATVDIQNPDITSSRYRGLPAPGPSPADKKRSGKKKISK ADIGAPSGFKHVSHVGWDPQNGFDVNNLDPDLRSLFSRAGISEAQLTDAETSKLIYDFIE DQGGLEAVRQEMRRQEPLPPPPPPSRGGNQLPRPPIVGGNKGRSGPLPPVPLGIAPPPPTP RGPPPPGRGGPPPPPPPATGRSGPLPPPPPGAGGPPMPPPPPPPPPPPSSGNGPAPPPLPPAL VPAGGLAPGGGRGALLDQIRQGIQLNKTPGAPES S ALQPPPQS SEGLVGALMHVMQKRS RAIHS SDEGEDQ AGDEDEDDEWDD SEQ ID NO: 101. WASL

[0368] MS S VQQQPPPPRRVTNVGSLLLTPQENESLFTFLGKKCVTMS S AVVQLYAADRNCMWS KKCSGVACLVKDNPQRSYFLRIFDIKDGKLLWEQELYNNFVYNSPRGYFHTFAGDTCQ VALNFANEEEAKKFRKAVTDLLGRRQRKSEKRRDPPNGPNLPMATVDIKNPEITTNRFY GPQVNNISHTKEKKKGKAKKKRLTKADIGTPSNFQHIGHVGWDPNTGFDLNNLDPELK NLFDMCGISEAQLKDRETSKVIYDFIEKTGGVEAVKNELRRQAPPPPPPSRGGPPPPPPPP HNSGPPPPPARGRGAPPPPPSRAPTAAPPPPPPSRPSVAVPPPPPNRMYPPPPPALPSSAPS GPPPPPP S VLGVGP VAPPPPPPPPPPPGPPPPPGLP SDGDHQ VPTTAGNKAALLDQIREGA QLKKVEQNSRPVSCSGRDALLDQIRQGIQLKSVADGQESTPPTPAPTSGIVGALMEVMQ KRSKAIHS SDEDEDEDDEEDFEDDDEWED SEQ ID NO: 102. WASF1

[0369] MPLVKRNIDPRHLCHTALPRGIKNELECVTNISLANIIRQLSSLSKYAEDIFGELFNEAHSF SFRVNSLQERVDRLSVSVTQLDPKEEELSLQDITMRKAFRSSTIQDQQLFDRKTLPIPLQE TYDVCEQPPPLNILTPYRDDGKEGLKFYTNPSYFFDLWKEKMLQDTEDKRKEKRKQKQ KNLDRPHEPEKVPRAPHDRRREWQKLAQGPELAEDDANLLHKHIEVANGPASHFETRP QTYVDHMDGS YSL S ALPF SQMSELLTRAEERVL VRPHEPPPPPPMHGAGD AKPIPTCIS S ATGLIENRPQSPATGRTPVFVSPTPPPPPPPLPSALSTSSLRASMTSTPPPPVPPPPPPPATAL QAPAVPPPPAPLQIAPGVLHPAPPPIAPPLVQPSPPVARAAPVCETVPVHPLPQGEVQGLP PPPPPPPLPPPGIRP S SP VTVT ALAHPP SGLHPTP ST APGPHVPLMPPSPP SQVIP ASEPKRH PSTLPVISDARSVLLEAIRKGIQLRKVEEQREQEAKHERIENDVATILSRRIAVEYSDSEDD SEFDEVDWLE

[0370] SEQ ID NO: 103. WASF2

[0371] MPLVTRNIEPRHLCRQTLPSVRSELECVTNITLANVIRQLGSLSKYAEDIFGELFTQANTF ASRVS SLAERVDRLQVKVTQLDPKEEEVSLQGINTRKAFRS STIQDQKLFDRNSLPVPVL ETYNTCDTPPPLNNLTPYRDDGKEALKFYTDPSYFFDLWKEKMLQDTKDIMKEKRKHR KEKKDNPNRGNVNPRKIKTRKEEWEKMKMGQEFVESKEKLGTSGYPPTLVYQNGSIGC VENVD AS S YPPPPQ SD S AS SPSP SF SEDNLPPPP AEFS YP VDNQRGSGLAGPKRS S VVSP S HPPPAPPLGSPPGPKPGFAPPPAPPPPPPPMIGIPPPPPPVGFGSPGTPPPPSPPSFPPHPDFAA PPPPPPPPAADYPTLPPPPLSQPTGGAPPPPPPPPPPGPPPPPFTGADGQPAIPPPLSDTTKPK SSLPAVSDARSDLLSAIRQGFQLRRVEEQREQEKRDVVGNDVATILSRRIAVEYSDSEDD SSEFDEDDWSD

[0372] SEQ ID NO: 104. WASF3

[0373] MPLVKRNIEPRHLCRGALPEGITSELECVTNSTLAAIIRQLSSLSKHAEDIFGELFNEANNF YIRANSLQDRIDRLAVKVTQLDSTVEEVSLQDINMKKAFKSSTVQDQQVVSKNSIPNPV ADIYNQSDKPPPLNILTPYRDDKKDGLKFYTDPSYFFDLWKEKMLQDTEDKRKEKRRQ KEQKRIDGTTREVKKVRKARNRRQEWNMMAYDKELRPDNRLSQSVYHGASSEGSLSP DTRSHASDVTDYSYPATPNHSLHPQPVTPSYAAGDVPPHGPASQAAEHEYRPPSASARH MALNRPQQPPPPPPPQAPEGSQASAPMAPADYGMLPAQIIEYYNPSGPPPPPPPPVIPSAQ TAF VSPLQMPMQPPFPAS AS STHAAPPHPPSTGLLVTAPPPPGPPPPPPGPPGPGS SLS SSP MHGPPVAEAKRQEPAQPPISDARSDLLAAIRMGIQLKKVQEQREQEAKREPVGNDVATI LSRRIAVEYSDSDDDSEFDENDWSD

[0374] SEQ ID NO: 105. SPIRE1

[0375] MAQAAGPAGGGEPRTEAVGGEGPREPGAAGGAAGGSRDALSLEEILRLYNQPINEEQA WAVCYQCCGSLRAAARRRQPRHRVRSAAQIRVWRDGAVTLAPAADDAGEPPPVAGKL GYSQCMETEVIESLGIIIYKALDYGLKENEERELSPPLEQLIDHMANTVEADGSNDEGYE AAEEGLGDEDEKRKISAIRSYRDVMKLCAAHLPTESDAPNHYQAVCRALFAETMELHT FLTKIKSAKENLKKIQEMEKSDESSTDLEELKNADWARFWVQVMRDLRNGVKLKKVQ ERQYNPLPIEYQLTPYEMLMDDIRCKRYTLRKVMVNGDIPPRLKKSAHEIILDFIRSRPPL NPVSARKLKPTPPRPRSLHERILEEIKAERKLRPVSPEEIRRSRLAMRPLSMSYSFDLSDVT TPESTKNLVESSMVNGGLTSQTKENGLSTSQQVPAQRKKLLRAPTLAELDSSESEEETLH KSTSS S S VSPSFPEEPVLEAVSTRKKPPKFLPIS STPQPERRQPPQRRHSIEKETPTNVRQFL PPSRQSSRSLEEFCYPVECLALTVEEVMHIRQVLVKAELEKYQQYKDIYTALKKGKLCF CCRTRRFSFFTWSYTCQFCKRPVCSQCCKKMRLPSKPYSTLPIFSLGPSALQRGESSMRS

[0376] EKPSTAHHRPLRSIARFSSKSKSMDKSDEELQFPKELMEDWSTMEVCVDCKKFISEIISSS RRSLVL ANKRARLKRKTQ SF YMS SPGP SE YCP SERTISEI

[0377] SEQ ID NO: 106. SPIRE2

[0378] MARAGSCGGAAAGAGRPEPWELSLEEVLKAYEQPLNEEQAWAVCFQGCRGLRGSPGR RLRDTGDLLLRGDGS VGAREPEAAEP ATMVVPLAS SE AQTVQ SLGF AIYRALDWGLDE SEERELSPQLERLIDLMANNDSEDSGCGAADEGYGGPEEEEEAEGVPRSVRTFAQAMRL CAARLTDPRGAQAHYQAVCRALFVETLELRAFLARVREAKEMLQKLREDEPHLETPRA ELDSLGHTDWARLWVQLMRELRRGVKLKKVQEQEFNPLPTEFQLTPFEMLMQDIRARN YKLRKVMVDGDIPPRVKKDAHELILDFIRSRPPLKQVSERRLRPLPPKQRSLHEKILEEIK QERRLRPVRGEGWAARGFGSLPCILNACSGDAKSTSCINLSVTDAGGSAQRPRPRVLLK APTLAEMEEMNTSEEEESPCGEVTLKRDRSFSEHDLAQLRSEVASGLQSATHPPGGTEPP RPRAGSAHVWRPGSRDQGTCPASVSDPSHPLLSNRGSSGDRPEASMTPDAKHLWLEFSH PVESLALTVEEVMDVRRVLVKAEMEKFLQNKELFS SLKKGKICCCCRAKFPLF SWPPSC LFCKRAVCTSCSIKMKMPSKKFGHIPVYTLGFESPQRVSAAKTAPIQRRDIFQSLQGPQW QSVEEAFPHIYSHGCVLKDVCSECTSFVADVVRSSRKSVDVLNTTPRRSRQTQSLYIPNT RTLDFK

[0379] SEQ ID NO: 107. Formin-1

[0380] MEGTHCTLQLHKPITELCYISFCLPKGEVRGFSYKGTVTLDRSNKGFHNCYQVREESDIIS LSQEPDEHPGDIFFKQTPTKDILTELYKLTTERERLLTNLLSSDHILGITMGNQEGKLQELS VSLAPEDDCFQSAGDWQGELPVGPLNKRSTHGNKKPRRSSGRRESFGALPQKRTKRKG RGGRESAPLMGKDKICSSHSLPLSRTRPNLWVLEEKGNLLPNGALACSLQRRESCPPDIP KTPDTDLGFGSFETAFKDTGLGREVLPPDCSSTEAGGDGIRRPPSGLEHQQTGLSESHQD PEKHPEAEKDEMEKPAKRTCKQKPVSKVVAKVQDLSSQVQRVVKTHSKGKETIAIRPA AHAEFVPKADLLTLPGAEAGAHGSRRQGKERQGDRSSQSPAGETASISSVSASAEGAVN KVPLKVIESEKLDEAPEGKRLGFPVHTSVPHTRPETRNKRRAGLPLGGHKSLFLDLPHKV GPDSSQPRGDKKKPSPPAPAALGKVFNNSASQSSTHKQTSPVPSPLSPRLPSPQQHHRILR LPALPGEREAALNDSPCRKSRVFSGCVSADTLEPPSSAKVTETKGASPAFLRAGQPRLVP GETLEKSLGPGKTTAEPQHQSPPGISSEGFPWDGFNEQTPKDLPNRDGGAWVLGYRAGP ACPFLLHEEREKSNRSELYLDLHPDHSLTEQDDRTPGRLQAVWPPPKTKDTEEKVGLKY TEAEYQAAILHLKREHKEEIENLQAQFELRAFHIRGEHAMITARLEETIENLKHELEHRW RGGCEERKDVCISTDDDCPPKTFRNVCVQTDRETFLKPCESESKTTRSNQLVPKKLNISS LSQLSPPNDHKDIHAALQPMEGMASNQQKALPPPPASIPPPPPLPSGLGSLSPAPPMPPVS AGPPLPPPPPPPPPLPPP S S AGPPPPPPPPPLPNSP APPNPGGPPP APPPPGLAPPPPPGLFFGL GSSSSQCPRKPAIEPSCPMKPLYWTRIQISDRSQNATPTLWDSLEEPDIRDPSEFEYLFSKD TTQQKKKPLSETYEKKNKVKKIIKLLDGKRSQTVGILISSLHLEMKDIQQAIFNVDDSVV DLETLAALYENRAQEDELVKIRKYYETSKEEELKLLDKPEQFLHELAQIPNFAERAQCIIF RSVFSEGITSLHRKVEIITRASKDLLHVKSVKDILALILAFGNYMNGGNRTRGQADGYSL EILPKLKDVKSRDNGINLVDYVVKYYLRYYDQEAGTEKSVFPLPEPQDFFLASQVKFED LIKDLRKLKRQLEASEKQMVVVCKESPKEYLQPFKDKLEEFFQKAKKEHKMEESHLEN AQKSFETTVRYFGMKPKSGEKEITPSYVFMVWYEFCSDFKTIWKRESKNISKERLKMAQ ESVSKLTSEKKVETKKINPTASLKERLRQKEASVTTN SEQ ID NO: 108. Formin-2

[0381] MGNQDGKLKRSAGDALHEGGGGAEDALGPRDVEATKKGSGGKKALGKHGKGGGGG GGGGESGKKKSKSDSRASVFSNLRIRKNLSKGKGAGGSREDVLDSQALQTGELDSAHSL LTKTPDLSLSADEAGLSDTECADPFEVTGPGGPGPAEARVGGRPIAEDVETAAGAQDGQ RTSSGSDTDIYSFHSATEQEDLLSDIQQAIRLQQQQQQQLQLQLQQQQQQQQLQGAEEP AAPPTAVSPQPGAFLGLDRFLLGPSGGAGEAPGSPDTEQALSALSDLPESLAAEPREPQQ PP SPGGLP VSEAP SLP AAQP AAKD SP S ST AFPFPEAGPGEEAAGAP VRGAGDTDEEGEED AFEDAPRGSPGEEWAPEVGEDAPQRLGEEPEEEAQGPDAPAAASLPGSPAPSQRCFKPY PLITPC YIKTTTRQLS SPNHSP SQ SPNQ SPRIKRRPEP SLSRGSRTAL AS VAAP AKKHRADG GLAAGLSRSADWTEELGARTPRVGGSAHLLERGVASDSGGGVSPALAAKASGAPAAA DGFQNVFTGRTLLEKLFSQQENGPPEEAEKFCSRIIAMGLLLPFSDCFREPCNQNAQTNA ASFDQDQLYTWAAVSQPTHSLDYSEGQFPRRVPSMGPPSKPPDEEHRLEDAETESQSAV SETPQKRSDAVQKEVVDMKSEGQATVIQQLEQTIEDLRTKIAELERQYPALDTEVASGH QGLENGVTASGDVCLEALRLEEKEVRHHRILEAKSIQTSPTEEGGVLTLPPVDGLPGRPP CPPGAESGPQTKFCSEISLIVSPRRISVQLDSHQPTQSISQPPPPPSLLWSAGQGQPGSQPPH SISTEFQTSHEHSVSSAFKNSCNIPSPPPLPCTESSSSMPGLGMVPPPPPPLPGMTVPTLPST AIPQPPPLQGTEMLPPPPPPLPGAGIPPPPPLPGAGILPLPPLPGAGIPPPPPLPGAAIPPPPPL PGAGIPLPPPLPGAGIPPPPPLPGAGIPPPPPLPGAGIPPPPPLPGAGIPPPPPLPGAGIPPPPPL PGAGIPPPPPLPGAGIPPPPPLPGAGIPPPPPLPGAGIPPPPPLPGAGIPPPPPLPGAGIPPPPPL PGAGIPPPPPLPGVGIPPPPPLPGAGIPPPPPLPGAGIPPPPPLPGAGIPPPPPLPRVGIPPPPPL PGAGIPPPPPLPGAGIPPPPPLPGVGIPPPPPLPGVGIPPPPPLPGAGIPPPPPLPGMGIPPAPA PPLPPPGTGIPPPPLLPVSGPPLLPQVGSSTLPTPQVCGFLPPPLPSGLFGLGMNQDKGSRK QPIEPCRPMKPLYWTRIQLHSKRDSSTSLIWEKIEEPSIDCHEFEELFSKTAVKERKKPISD TISKTKAKQVVKLLSNKRSQAVGILMSSLHLDMKDIQHAVVNLDNSVVDLETLQALYE NRAQ SDELEKIEKHGRS SKDKENAKSLDKPEQFL YEL SLIPNF SERVFCILFQ STF SESIC SI RRKLELLQKLCETLKNGPGVMQVLGLVLAFGNYMNGGNKTRGQADGFGLDILPKLKD VKSSDNSRSLLSYIVSYYLRNFDEDAGKEQCLFPLPEPQDLFQASQMKFEDFQKDLRKL KKDLKACEVEAGKVYQVSSKEHMQPFKENMEQFIIQAKIDQEAEENSLTETHKCFLETT AYFFMKPKLGEKEVSPNAFFSIWHEFSSDFKDFWKKENKLLLQERVKEAEEVCRQKKG

[0382] KSLYKIKPRHDSGIKAKISMKT

[0383] SEQ ID NO: 109. SCAR3

[0384] MKVRSAGGDGDALCVTEEDLAGDDEDMPTFPCTQKGRPGPRCSRCQKNLSLHTSVRIL YLFLALLLVAVA VLASLVFRKVD SL SEDISLTQ SIYDKKLVLMQKNLQGLDPKALNNC S FCHEAGQLGPEIRKLQEELEGIQKLLLAQEVQLDQTLQAQEVLSTTSRQISQEMGSCSFSI HQVNQSLGLFLAQVRGWQATTAGLDLSLKDLTQECYDVKAAVHQINFTVGQTSEWIH GIQRKTDEETLTLQKIVTDWQNYTRLFSGLRTTSTKTGEAVKNIQATLGASSQRISQNSE SMHDL VLQ VMGLQLQLDNIS SFLDDHEENMHDLQ YHTHYAQNRT VERFE SLEGRMAS HEIEIGTIFTNINATDNHVHSMLKYLDDVRLSCTLGFHTHAEELYYLNKSVSIMLGTTDL LRERFSLLSARLDLNVRNLSMIVEEMKAVDTQHGEILRNVTILRGAPGPPGPRGFKGDM GVKGPVGGRGPKGDPGSLGPLGPQGPQGQPGEAGPVGERGPVGPRGFPGLKGSKGSFG TGGPRGQPGPKGDIGPPGPEGPPGSPGPSGPQGKPGIAGKTGSPGQRGAMGPKGEPGIQG PPGLPGPPGPPGSQSFY SEQ ID NO: 110. SCAR5

[0385] MENKAMYLHTVSDCDTSSICEDSFDGRSLSKLNLCEDGPCHKRRASICCTQLGSLSALK HAVLGLYLLVFLILVGIFILAVSRPRSSPDDLKALTRNVNRLNESFRDLQLRLLQAPLQA DLTEQVWKVQDALQNQSDSLLALAGAVQRLEGALWGLQAQAVQTEQAVALLRDRTG QQSDTAQLELYQLQVESNSSQLLLRRHAGLLDGLARRVGILGEELADVGGVLRGLNHSL SYDVALHRTRLQDLRVLVSNASEDTRRLRLAHVGMELQLKQELAMLNAVTEDLRLKD WEHSIALRNISLAKGPPGPKGDQGDEGKEGRPGIPGLPGLRGLPGERGTPGLPGPKGDDG KLGATGPMGMRGFKGDRGPKGEKGEKGDRAGDASGVEAPMMIRLVNGSGPHEGRVE VYHDRRWGTVCDDGWDKKDGDVVCRMLGFRGVEEVYRTARFGQGTGRIWMDDVAC KGTEETIFRCSFSKWGVTNCGHAEDASVTCNRH

[0386] SEQ ID NO: 111. Myosin light chain kinase

[0387] MGD VKL VAS SHISKT SLS VDP SRVD SMPLTEAP AFILPPRNLCIKEGAT AKFEGRVRGYP EPQVTWHRNGQPITSGGRFLLDCGIRGTFSLVIHAVHEEDRGKYTCEATNGSGARQVTV ELTVEGSF AKQLGQP VVSKTLGDRF S AP AVETRP SIWGECPPKF ATKLGRVVVKEGQMG RFSCKITGRPQPQVTWLKGNVPLQPSARVSVSEKNGMQVLEIHGVNQDDVGVYTCLVV NGSGKASMS AELSIQGLD S ANRSF VRETKATNSD VRKE VTNVISKE SKLD SLEAAAK SK NC S SPQRGGSPPWAANSQPQPPRESKLESCKD SPRTAPQTP VLQKT S S SITLQ AARVQPEP RAPGLGVLSPSGEERKRPAPPRPATFPTRQPGLGSQDVVSKAANRRIPMEGQRDSAFPKF ESKPQSQEVKENQTVKFRCEVSGIPKPEVAWFLEGTPVRRQEGSIEVYEDAGSHYLCLL KARTRDSGTYSCTASNAQGQLSCSWTLQVERLAVMEVAPSFSSVLKDCAVIEGQDFVL QCSVRGTPVPRITWLLNGQPIQYARSTCEAGVAELHIQDALPEDHGTYTCLAENALGQV SCSAWVTVHEKKSSRKSEYLLPVAPSKPTAPIFLQGLSDLKVMDGSQVTMTVQVSGNPP PEVIWLHNGNEIQESEDFHFEQRGTQHSLCIQEVFPEDTGTYTCEAWNSAGEVRTQAVL TVQEPHDGTQPWFISKPRSVTASLGQSVLISCAIAGDPFPTVHWLRDGKALCKDTGHFE VLQNEDVFTLVLKKVQPWHAGQYEILLKNRVGECSCQVSLMLQNSSARALPRGREPAS CEDLCGGGVGADGGGSDRYGSLRPGWPARGQGWLEEEDGEDVRGVLKRRVETRQHTE EAIRQQEVEQLDFRDLLGKKVSTKTLSEDDLKEIPAEQMDFRANLQRQVKPKTVSEEER KVHSPQQVDFRSVLAKKGTSKTPVPEKVPPPKPATPDFRSVLGGKKKLPAENGSSSAET LNAKAVESSKPLSNAQPSGPLKPVGNAKPAETLKPMGNAKPAETLKPMGNAKPDENLK SASKEELKKDVKNDVNCKRGHAGTTDNEKRSESQGTAPAFKQKLQDVHVAEGKKLLL QCQVS SDPPATIIWTLNGKTLKTTKFIILSQEGSLC S VSIEKALPEDRGLYKC VAKND AGQ AECSCQVTVDDAPASENTKAPEMKSRRPKSSLPPVLGTESDATVKKKPAPKTPPKAAMP PQIIQFPEDQKVRAGESVELFGKVTGTQPITCTWMKFRKQIQESEHMKVENSENGSKLTI LAARQEHCGCYTLLVENKLGSRQAQVNLTVVDKPDPPAGTPCASDIRSSSLTLSWYGSS YDGGSAVQSYSIEIWDSANKTWKELATCRSTSFNVQDLLPDHEYKFRVRAINVYGTSEP SQESELTTVGEKPEEPKDEVEVSDDDEKEPEVDYRTVTINTEQKVSDFYDIEERLGSGKF GQVFRLVEKKTRKVWAGKFFKAYSAKEKENIRQEISIMNCLHHPKLVQCVDAFEEKANI VMVLEIVSGGELFERIIDEDFELTERECIKYMRQISEGVEYIHKQGIVHLDLKPENIMCVN KTGTRIKLIDFGLARRLENAGSLKVLFGTPEFVAPEVINYEPIGYATDMWSIGVICYILVS GLSPFMGDNDNETLANVTSATWDFDDEAFDEISDDAKDFISNLLKKDMKNRLDCTQCL QHPWLMKDTKNMEAKKLSKDRMKKYMARRKWQKTGNAVRAIGRLSSMAMISGLSG RKSSTGSPTSPLNAEKLESEEDVSQAFLEAVAEEKPHVKPYFSKTIRDLEVVEGSAARFD CKIEGYPDPEVVWFKDDQSIRESRHFQIDYDEDGNCSLIISDVCGDDDAKYTCKAVNSL

[0388] GEATCTAELIVETMEEGEGEGEEEEE

[0389] SEQ ID NO: 112. Myosin light chain kinase 3

[0390] MSGTSKESLGHGGLPGLGKTCLTTMDTKLNMLNEKVDQLLHFQEDVTEKLQSMCRDM GHLERGLHRLEASRAPGPGGADGVPHIDTQAGWPEVLELVRAMQQDAAQHGARLEAL FRMVAAVDRAIALVGATFQKSKVADFLMQGRVPWRRGSPGDSPEENKERVEEEGGKP KHVLSTSGVQSDAREPGEESQKADVLEGTAERLPPIRASGLGADPAQAVVSPGQGDGVP GPAQAFPGHLPLPTKVEAKAPETPSENLRTGLELAPAPGRVNVVSPSLEVAPGAGQGAS SSRPDPEPLEEGTRLTPGPGPQCPGPPGLPAQARATHSGGETPPRISIHIQEMDTPGEMLM TGRGSLGPTLTTEAPAAAQPGKQGPPGTGRCLQAPGTEPGEQTPEGARELSPLQES S SPG GVKAEEEQRAGAEPGTRPSLARSDDNDHEVGALGLQQGKSPGAGNPEPEQDCAARAPV RAEAVRRMPPGAEAGSVVLDDSPAPPAPFEHRVVSVKETSISAGYEVCQHEVLGGGRFG QVHRCTEKSTGLPLAAKIIKVKSAKDREDVKNEINIMNQLSHVNLIQLYDAFESKHSCTL VMEYVDGGELFDRITDEKYHLTELDVVLFTRQICEGVHYLHQHYILHLDLKPENILCVN QTGHQIKIIDFGLARRYKPREKLKVNFGTPEFLAPEVVNYEFVSFPTDMWSVGVITYMLL SGLSPFLGETDAETMNFIVNCSWDFDADTFEGLSEEAKDFVSRLLVKEKSCRMSATQCL KHEWLNNLPAKASRSKTRLKSQLLLQKYIAQRKWKKHFYVVTAANRLRKFPTSP SEQ ID NO: 114. Unconventional myosin-lg

[0391] MEDEEGPEYGKPDFVLLDQVTMEDFMRNLQLRFEKGRIYTYIGEVLVSVNPYQELPLYG PEAIARYQGRELYERPPHLYAVANAAYKAMKHRSRDTCIVISGESGAGKTEASKHIMQY IAAVTNPSQRAEVERVKDVLLKSTCVLEAFGNARTNRNHNSSRFGKYMDINFDFKGDPI GGHIHSYLLEKSRVLKQHVGERNFHAFYQLLRGSEDKQLHELHLERNPAVYNFTHQGA GLNMTVHS ALD SDEQ SHQ AVTE AMRVIGF SPEEVES VHRILAAILHLGNIEF VETEEGGL QKEGLAVAEEALVDHVAELTATPRDLVLRSLLARTVASGGRELIEKGHTAAEASYARD ACAKAVYQRLFEWVVNRINSVMEPRGRDPRRDGKDTVIGVLDIYGFEVFPVNSFEQFCI NYCNEKLQQLFIQLILKQEQEEYEREGITWQSVEYFNNATIVDLVERPHRGILAVLDEAC SSAGTITDRIFLQTLDMHHRHHLHYTSRQLCPTDKTMEFGRDFRIKHYAGDVTYSVEGFI DKNRDFLFQDFKRLLYNSTDPTLRAMWPDGQQDITEVTKRPLTAGTLFKNSMVALVEN LASKEPFYVRCIKPNEDKVAGKLDENHCRHQVAYLGLLENVRVRRAGFASRQPYSRFL LRYKMTCEYTWPNHLLGSDKAAVSALLEQHGLQGDVAFGHSKLFIRSPRTLVTLEQSR ARLIPIIVLLLQKAWRGTLARWRCRRLRAIYTIMRWFRRHKVRAHLAELQRRFQAARQP PLYGRDLVWPLPPAVLQPFQDTCHALFCRWRARQLVKNIPPSDMPQIKAKVAAMGALQ GLRQDWGCRRAW ARD YL S S ATDNPT AS SLF AQRLKTLQDKDGFGA VLF S SHVRK VNR FHKIRNRALLLTDQHLYKLDPDRQYRVMRAVPLEAVTGLSVTSGGDQLVVLHARGQD DLVVCLHRSRPPLDNRVGELVGVLAAHCQGEGRTLEVRVSDCIPLSHRGVRRLISVEPRP EQPEPDFRC ARGSFTLLWP SR SEQ ID NO: 115. My osin-1

[0392] MSSDSEMAIFGEAAPFLRKSERERIEAQNKPFDAKTSVFVVDPKESFVKATVQSREGGK VTAKTEAGATVTVKDDQVFPMNPPKYDKIEDMAMMTHLHEPAVLYNLKERYAAWMI YTYSGLFCVTVNPYKWLPVYNAEVVTAYRGKKRQEAPPHIFSISDNAYQFMLTDRENQ SILITGESGAGKTVNTKRVIQYFATIAVTGEKKKEEVTSGKMQGTLEDQIISANPLLEAFG NAKTVRNDNSSRFGKFIRIHFGTTGKLASADIETYLLEKSRVTFQLKAERSYHIFYQIMSN KKPDLIEMLLITTNPYDYAFVSQGEITVPSIDDQEELMATDSAIEILGFTSDERVSIYKLTG AVMHYGNMKFKQKQREEQAEPDGTEVADKAAYLQNLNSADLLKALCYPRVKVGNEY VTKGQTVQQVYNAVGALAKAVYDKMFLWMVTRINQQLDTKQPRQYFIGVLDIAGFEIF DFNSLEQLCINFTNEKLQQFFNHHMFVLEQEEYKKEGIEWTFIDFGMDLAACIELIEKPM GIFSILEEECMFPKATDTSFKNKLYEQHLGKSNNFQKPKPAKGKPEAHFSLIHYAGTVDY NIAGWLDKNKDPLNETVVGLYQKSAMKTLALLFVGATGAEAEAGGGKKGGKKKGSSF QTVSALFRENLNKLMTNLRSTHPHFVRCIIPNETKTPGAMEHELVLHQLRCNGVLEGIRI CRKGFPSRILYADFKQRYKVLNASAIPEGQFIDSKKASEKLLGSIDIDHTQYKFGHTKVFF KAGLLGLLEEMRDEKLAQLITRTQAMCRGFLARVEYQKMVERRESIFCIQYNVRAFMN VKHWPWMKLYFKIKPLLKSAETEKEMANMKEEFEKTKEELAKTEAKRKELEEKMVTL MQEKNDLQLQVQAEADSLADAEERCDQLIKTKIQLEAKIKEVTERAEDEEEINAELTAK KRKLEDECSELKKDIDDLELTLAKVEKEKHATENKVKNLTEEMAGLDETIAKLTKEKK ALQEAHQQTLDDLQAEEDKVNTLTKAKIKLEQQVDDLEGSLEQEKKIRMDLERAKRKL EGDLKLAQESTMDIENDKQQLDEKLKKKEFEMSGLQSKIEDEQALGMQLQKKIKELQA RIEELEEEIEAERASRAKAEKQRSDL SRELEEISERLEEAGGAT S AQ I EM N K K RE AEFQKM RRDLEEATLQHEATAATLRKKHADSVAELGEQIDNLQRVKQKLEKEKSEMKMEIDDLA SNMETVSK AKGNLEKMCRALEDQL SEIKTKEEEQQRLINDLT AQRARLQTE SGEYSRQL DEKDTLVSQLSRGKQAFTQQIEELKRQLEEEIKAKSALAHALQSSRHDCDLLREQYEEE QEAKAELQRAMSKANSEVAQWRTKYETDAIQRTEELEEAKKKLAQRLQDAEEHVEAV NAKCASLEKTKQRLQNEVEDLMIDVERTNAACAALDKKQRNFDKILAEWKQKCEETH AELEASQKESRSLSTELFKIKNAYEESLDQLETLKRENKNLQQEISDLTEQIAEGGKRIHE LEKIKKQVEQEKSELQAALEEAEASLEHEEGKILRIQLELNQVKSEVDRKIAEKDEEIDQ MKRNHIRIVESMQSTLDAEIRSRNDAIRLKKKMEGDLNEMEIQLNHANRMAAEALRNY RNTQAILKDTQLHLDDALRSQEDLKEQLAMVERRANLLQAEIEELRATLEQTERSRKIA EQELLDASERVQLLHTQNTSLINTKKKLETDISQIQGEMEDIIQEARNAEEKAKKAITDA AMMAEELKKEQDTSAHLERMKKNLEQTVKDLQHRLDEAEQLALKGGKKQIQKLEARV RELEGEVESEQKRNVEAVKGLRKHERKVKELTYQTEEDRKNILRLQDLVDKLQAKVKS YKRQ AEE AEEQ SNVNL SKFRRIQHELEE AEERADIAE S Q VNKLR VK SRE VHTKIISEE SEQ ID NO: 116. Myosin-2

[0393] MSSDSELAVFGEAAPFLRKSERERIEAQNRPFDAKTSVFVAEPKESFVKGTIQSREGGKV TVKTEGGATLTVKDDQVFPMNPPKYDKIEDMAMMTHLHEPAVLYNLKERYAAWMIYT YSGLFCVTVNPYKWLPVYKPEVVTAYRGKKRQEAPPHIFSISDNAYQFMLTDRENQSILI TGESGAGKTVNTKRVIQYFATIAVTGEKKKEEITSGKIQGTLEDQIISANPLLEAFGNAKT VRNDNSSRFGKFIRIHFGTTGKLASADIETYLLEKSRVVFQLK AF.RSYHIFYQITSNKKPF. LIEMLLITTNPYDYPFVSQGEISVASIDDQEELMATDSAIDILGFTNEEKVSIYKLTGAVM HYGNLKFKQKQREEQAEPDGTEVADKAAYLQSLNSADLLKALCYPRVKVGNEYVTKG QTVEQVSNAVGALAKAVYEKMFLWMVARINQQLDTKQPRQYFIGVLDIAGFEIFDFNS LEQLCINFTNEKLQQFFNHHMFVLEQEEYKKEGIEWTFIDFGMDLAACIELIEKPMGIFSI LEEECMFPKATDTSFKNKLYDQHLGKSANFQKPKVVKGKAEAHFALIHYAGVVDYNIT GWLEKNKDPLNETVVGLYQKSAMKTLAQLFSGAQTAEGEGAGGGAKKGGKKKGSSF QTVSALFRENLNKLMTNLRSTHPHFVRCIIPNETKTPGAMEHELVLHQLRCNGVLEGIRI CRKGFPSRILYADFKQRYKVLNASAIPEGQFIDSKKASEKLLASIDIDHTQYKFGHTKVFF KAGLLGLLEEMRDDKLAQLITRTQARCRGFLARVEYQRMVERREAIFCIQYNIRSFMNV KHWPWMKLFFKIKPLLKSAETEKEMATMKEEFQKIKDELAKSEAKRKELEEKMVTLLK EKNDLQLQVQAEAEGLADAEERCDQLIKTKIQLEAKIKEVTERAEDEEEINAELTAKKR KLEDECSELKKDIDDLELTLAKVEKEKHATENKVKNLTEEMAGLDETIAKLTKEKKALQ EAHQQTLDDLQAEEDKVNTLTKAKIKLEQQVDDLEGSLEQEKKLRMDLERAKRKLEGD LKLAQESIMDIENEKQQLDEKLKKKEFEISNLQSKIEDEQALGIQLQKKIKELQARIEELE EEIEAERASRAKAEKQRSDLSRELEEISERLEEAGGATSAQIEMNKKREAEFQKMRRDLE EATLQHEATAATLRKKHADSVAELGEQIDNLQRVKQKLEKEKSEMKMEIDDLASNVET VSKAKGNLEKMCRTLEDQLSELKSKEEEQQRLINDLTAQRGRLQTESGEFSRQLDEKEA LVSQLSRGKQAFTQQIEELKRQLEEEIKAKNALAHALQSSRHDCDLLREQYEEEQESKA ELQRALSKANTEVAQWRTKYETDAIQRTEELEEAKKKLAQRLQAAEEHVEAVNAKCAS LEKTKQRLQNEVEDLMLDVERTNAACAALDKKQRNFDKILAEWKQKCEETHAELEAS QKEARSLGTELFKIKNAYEESLDQLETLKRENKNLQQEISDLTEQIAEGGKRIHELEKIKK QVEQEKCELQAALEEAEASLEHEEGKILRIQLELNQVKSEVDRKIAEKDEEIDQLKRNHI RIVESMQSTLDAEIRSRNDAIRLKKKMEGDLNEMEIQLNHANRMAAEALRNYRNTQGIL KDTQIHLDDALRSQEDLKEQLAMVERRANLLQAEIEELRATLEQTERSRKIAEQELLDAS ERVQLLHTQNTSLINTKKKLETDISQMQGEMEDILQEARNAEEKAKKAITDAAMMAEE LKKEQDTSAHLERMKKNMEQTVKDLQLRLDEAEQLALKGGKKQIQKLEARVRELEGE VESEQI<RNAEAVI<GLRKHERRVI<ELTYQTEEDRI<NILRLQDLVDI<LQAI<VI<SYI<RQAE EAEEQ SNTN LAK FRK LQHELEE AEER A D I AE SQ VNKLRVKSREVHTKVISEE SEQ ID NO: 117. Unconventional myosin-Va MAASELYTKFARVWIPDPEEVWKSAELLKDYKPGDKVLLLF1LEEGKDLEYHLDPKTKE LPHLRNPDILVGENDLTALSYLHEPAVLHNLRVRFIDSKLIYTYCGIVLVAINPYEQLPIY GEDIINAYSGQNMGDMDPHIFAVAEEAYKQMARDERNQSIIVSGESGAGKTVSAKYAM RYFATVSGSASEANVEEKVLASNPIMESIGNAKTTRNDNSSRFGKYIEIGFDKRYRIIGAN MRTYLLEKSRVVFQAEEERNYHIFYQLCASAKLPEFKMLRLGNADNFNYTKQGGSPVIE GVDDAKEMAHTRQACTLLGISESHQMGIFRILAGILHLGNVGFTSRDADSCTIPPKHEPL CIFCELMGVDYEEMCHWLCHRKLATATETYIKPISKLQATNARDALAKHIYAKLFNWIV DNVNQALHSAVKQHSFIGVLDIYGFETFEINSFEQFCINYANEKLQQQFNMHVFKLEQEE YMKEQIPWTLIDFYDNQPCINLIESKLGILDLLDEECKMPKGTDDTWAQKLYNTHLNKC ALFEKPRLSNKAFIIQHFADKVEYQCEGFLEKNKDTVFEEQIKVLKSSKFKMLPELFQDD EKAISPTSATSSGRTPLTRTPAKPTKGRPGQMAKEHKKTVGHQFRNSLHLLMETLNATT PHYVRCIKPNDFKFPFTFDEKRAVQQLRACGVLETIRISAAGFPSRWTYQEFFSRYRVLM KQKDVLSDRKQTCKNVLEKLILDKDKYQFGKTKIFFRAGQVAYLEKLRADKLRAACIRI QKTIRGWLLRKKYLRMRKAAITMQRYVRGYQARCYAKFLRRTKAATIIQKYWRMYVV RRRYI<IRRAATIVLQSYLRGFLARNRYRKILREHKAVIIQI<RVRGWLARTHYI<RSMHAII

[0394] YLQCCFRRMMAKRELKKLKIEARSVERYKKLHIGMENKIMQLQRKVDEQNKDYKCLV EKLTNLEGIYNSETEKLRSDLERLQLSEEEAKVATGRVLSLQEEIAKLRKDLEQTRSEKK CIEEHADRYKQETEQLVSNLKEENTLLKQEKEALNHRIVQQAKEMTETMEKKLVEETK QLELDLNDERLRYQNLLNEF SRLEERYDDLKEEMTLMVHVPKPGHKRTD STHS SNESEY IFSSEIAEMEDIPSRTEEPSEKKVPLDMSLFLKLQKRVTELEQEKQVMQDELDRKEEQVL RSKAKEEERPQIRGAELEYESLKRQELESENKKLKNELNELRKALSEKSAPEVTAPGAPA YRVLMEQLTSVSEELDVRKEEVLILRSQLVSQKEAIQPKDDKNTMTDSTILLEDVQKMK DKGEIAQAYIGLKETNRSSALDYHELNEDGELWLVYEGLKQANRLLESQLQSQKRSHE NEAEALRGEIQSLKEENNRQQQLLAQNLQLPPEARIEASLQHEITRLTNENLDLMEQLEK QDKTVRKLKKQLKVFAKKIGELEVGQMENISPGQIIDEPIRPVNIPRKEKDFQGMLEYKK EDEQKLVKNLILELKPRGVAVNLIPGLPAYILFMCVRHADYLNDDQKVRSLLTSTINSIK KVLKKRGDDFETVSFWLSNTCRFLHCLKQYSGEEGFMKHNTSRQNEHCLTNFDLAEYR QVLSDLAIQIYQQLVRVLENILQPMIVSGMLEHETIQGVSGVKPTGLRKRTSSIADEGTYT LDSILRQLNSFHSVMCQHGMDPELIKQVVKQMFYIIGAITLNNLLLRKDMCSWSKGMQI RYNVSQLEEWLRDKNLMNSGAKETLEPLIQAAQLLQVKKKTDDDAEAICSMCNALTTA QIVKVLNLYTPVNEFEERVSVSFIRTIQMRLRDRKDSPQLLMDAKHIFPVTFPFNPSSLAL ETIQIPASLGLGFISRV

[0395] SEQ ID NO: 118. Unconventional myosin-Vb

[0396] MSVGELYSQCTRVWIPDPDEVWRSAELTKDYKEGDKSLQLRLEDETILEYPIDVQRNQL PFLRNPDILVGENDLTALSYLHEPAVLHNLKVRFLESNHIYTYCGIVLVAINPYEQLPIYG QDVIYTYSGQNMGDMDPHIFAVAEEAYKQMARDEKNQSIIVSGESGAGKTVSAKYAM RYF ATVGGS ASETNIEEKVLAS SPIME AIGNAKTTRNDNS SRFGKYIQIGFDKRYHIIGAN MRTYLLEKSRVVFQADDERNYHIFYQLCAAAGLPEFKELALTSAEDFFYTSQGGDTSIE GVDDAEDFEKTRQAFTLLGVKESHQMSIFKIIASILHLGSVAIQAERDGDSCSISPQDVYL SNFCRLLGVEHSQMEHWLCHRKLVTTSETYVKTMSLQQVINARNALAKHIYAQLFGWI VEHINKALHTSLKQHSFIGVLDIYGFETFEVNSFEQFCINYANEKLQQQFNSHVFKLEQEE YMKEQIPWTLIDFYDNQPCIDLIEAKLGILDLLDEECKVPKGTDQNWAQKLYDRHSSSQ HFQKPRMSNTAFIIVHFADKVEYLSDGFLEKNRDTVYEEQINILKASKFPLVADLFHDDK DPVPATTPGKGS S SKIS VRS ARPPMKVSNKEHKKTVGHQFRTSLHLLMETLNATTPHYV RCII<PNDEKLPFHFDPKRAVQQLRACGVLETIRISAAGYPSRWAYHDFFNRYRVLVI<I<R ELANTDKKAICRSVLENLIKDPDKFQFGRTKIFFRAGQVAYLEKLRADKFRTATIMIQKT VRGWLQKVKYHRLKGATLTLQRYCRGHLARRLAEHLRRIRAAVVLQKHYRMQRARQ AYQRVRRAAVVIQAFTRAMFVRRTYRQVLMEHKATTIQKHVRGWMARRHFQRLRDA AIVIQCAFRMLKARRELKALRIEARSAEHLKRLNVGMENKVVQLQRKIDEQNKEFKTLS EQLSVTTSTYTMEVERLKKELVHYQQSPGEDTSLRLQEEVESLRTELQRAHSERKILEDA HSREKDELRKRVADLEQENALLKDEKEQLNNQILCQSKDEFAQNSVKENLMKKELEEE RSRYQNLVKEYSQLEQRYDNLRDEMTIIKQTPGHRRNPSNQSSLESDSNYPSISTSEIGDT EDALQQVEEIGLEKAAMDMTVFLKLQKRVRELEQERKKLQVQLEKREQQDSKKVQAE PPQTDIDLDPNADLAYNSLKRQELESENKKLKNDLNELRKAVADQATQNNSSHGSPDS YSLLLNQLKLAHEELEVRKEEVLILRTQIVSADQRRLAGRNAEPNINARSSWPNSEKHVD QEDAIEAYHGVCQTNSKTEDWGYLNEDGELGLAYQGLKQVARLLEAQLQAQSLEHEE EVEHLKAQLEALKEEMDKQQQTFCQTLLLSPEAQVEFGVQQEISRLTNENLDLKELVEK LEKNERKLKKQLKIYMKKAQDLEAAQALAQSERKRHELNRQVTVQRKEKDFQGMLEY HKEDEALLIRNLVTDLKPQMLSGTVPCLPAYILYMCIRHADYTNDDLKVHSLLTSTINGI KKVLKKHNDDFEMTSFWLSNTCRLLHCLKQYSGDEGFMTQNTAKQNEHCLKNFDLTE YRQVLSDLSIQIYQQLIKIAEGVLQPMIVSAMLENESIQGLSGVKPTGYRKRSSSMADGD NSYCLEAIIRQMNAFHTVMCDQGLDPEIILQVFKQLFYMINAVTLNNLLLRKDVCSWST GMQLRYNISQLEEWLRGRNLHQSGAVQTMEPLIQAAQLLQLKKKTQEDAEAICSLCTSL STQQIVKILNLYTPLNEFEERVTVAFIRTIQAQLQERNDPQQLLLDAKHMFPVLFPFNPSS LTMD SIH IP ACLNLEFLNEV

[0397] SEQ ID NO: 119. Unconventional myosin- VI

[0398] MEDGKPVWAPHPTDGFQMGNIVDIGPDSLTIEPLNQKGKTFLALINQVFPAEEDSKKDV EDNC SLMYLNEATLLHNIKVRYSKDRIYTYVANILIAVNPYFDIPKIYS SEAIKS YQGKSL GTRPPHVFAIADKAFRDMKVLKMSQSIIVSGESGAGKTENTKFVLRYLTESYGTGQDID DRIVEANPLLEAFGNAKTVRNNNS SRFGKF VEIHFNEKS S VVGGF VSHYLLEKSRIC VQG KEERNYHIFYRLCAGASEDIREKLHLSSPDNFRYLNRGCTRYFANKETDKQILQNRKSPE YLKAGSMKDPLLDDHGDFIRMCTAMKKIGLDDEEKLDLFRVVAGVLHLGNIDFEEAGS TSGGCNLKNKSAQSLEYCAELLGLDQDDLRVSLTTRVMLTTAGGTKGTVIKVPLKVEQ ANNARDALAKTVYSHLFDHVVNRVNQCFPFETSSYFIGVLDIAGFEYFEHNSFEQFCINY CNEKLQQFFNERILKEEQELYQKEGLGVNEVHYVDNQDCIDLIEAKLVGILDILDEENRL PQPSDQHFTSAVHQKHKDHFRLTIPRKSKLAVHRNIRDDEGFIIRHFAGAVCYETTQFVE KNNDALHMSLESLICESRDKFIRELFESSTNNNKDTKQKAGKLSFISVGNKFKTQLNLLL DKLRSTGASFIRCIKPNLKMTSHHFEGAQILSQLQCSGMVSVLDLMQGGYPSRASFHEL YNMYKKYMPDKLARLDPRLFCKALFKALGLNENDYKFGLTI<VFFRPGI<FAEFDQIMI<S DPDHLAELVKRVNHWLTCSRWKKVQWCSLSVIKLKNKIKYRAEACIKMQKTIRMWLC KRRHKPRJDGLVKVGTLKKRLDKFNEVVSVLKDGKPEMNKQIKNLEISIDTLMAKIKST MMTQEQIQKEYDALVKSSEELLSALQKKKQQEEEAERLRRIQEEMEKERKRREEDEKR RRKEEEERRMKLEMEAKRKQEEEERKKREDDEKRIQAEVEAQLARQKEEESQQQAVLE QERRDRELALRIAQSEAELISDEAQADLALRRSLDSYPVSKNDGTRPKMTPEQMAKEMS EFLSRGPAVLATKAAAGTKKYDLSKWKYAELRDTINTSCDIELLAACREEFHRRLKVYH AWKSKNKKRNTETEQRAPKSVTDYDFAPFLNNSPQQNPAAQIPARQREIEMNRQQRFFR IPFIRPADQYKDPQSKKKGWWYAHFDGPWIARQMELHPDKPPILLVAGKDDMEMCELN LEETGLTRKRGAEILPRQFEEIWERCGGIQYLQNAIESRQARPTYATAMLQSLLK JAK STAT signaling SEQ ID NO: 120. JAK1

[0399] MQYLNIKEDCNAMAFCAKMRSSKKTEVNLEAPEPGVEVIFYLSDREPLRLGSGEYTAEE LCIRAAQACRISPLCHNLFALYDENTKLWYAPNRTITVDDKMSLRLHYRMRFYFTNWH GTNDNEQSVWRHSPKKQKNGYEKKKIPDATPLLDASSLEYLFAQGQYDLVKCLAPIRD PKTEQDGHDIENECLGMAVLAISHYAMMKKMQLPELPKDISYKRYIPETLNKSIRQRNL LTRMRINNVFKDFLKEFNNKTICDSSVSTHDLKVKYLATLETLTKHYGAEIFETSMLLISS ENEMNWFHSNDGGNVLYYEVMVTGNLGIQWRHKPNVVSVEKEKNKLKRKKLENKHK KDEEKNKIREEWNNFSYFPEITHIVIKESVVSINKQDNKKMELKLSSHEEALSFVSLVDG YFRLTADAHHYLCTDVAPPLIVHNIQNGCHGPICTEYAINKLRQEGSEEGMYVLRWSCT DFDNILMTVTCFEKSEQVQGAQKQFKNFQIEVQKGRYSLHGSDRSFPSLGDLMSHLKKQ

[0400] ILRTDNISFMLKRCCQPKPREISNLLVATKKAQEWQPVYPMSQLSFDRILKKDLVQGEHL GRGTRTHIYSGTLMDYKDDEGTSEEKKIKVILKVLDPSHRDISLAFFEAASMMRQVSHK HIVYLYGVCVRDVENIMVEEFVEGGPLDLFMHRKSDVLTTPWKFKVAKQLASALSYLE DKDLVHGNVCTKNLLLAREGIDSECGPFIKLSDPGIPITVLSRQECIERIPWIAPECVEDSK NLSVAADKWSFGTTLWEICYNGEIPLKDKTLIEKERFYESRCRPVTPSCKELADLMTRC MNYDPNQRPFFRAIMRDINKLEEQNPDIVSEKKPATEVDPTHFEKRFLKRIRDLGEGHFG KVELCRYDPEGDNTGEQVAVKSLKPESGGNHIADLKKEIEILRNLYHENIVKYKGICTED GGNGIKLIMEFLPSGSLKEYLPKNKNKINLKQQLKYAVQICKGMDYLGSRQYVHRDLA ARNVL VESEHQ VKIGDFGLTKAIETDKEYYT VKDDRD SP VF WYAPECLMQ SKF YIASD V

[0401] WSFGVTLHELLTYCDSDSSPMALFLKMIGPTHGQMTVTRLVNTLKEGKRLPCPPNCPDE VYQLMRKCWEFQPSNRTSFQNLIEGFEALLK

[0402] SEQ ID NO: 12E JAK2

[0403] MGMACLTMTEMEGTSTSSIYQNGDISGNANSMKQIDPVLQVYLYHSLGKSEADYLTFPS GEYVAEEICIAASKACGITPVYHNMFALMSETERIWYPPNHVFHIDESTRHNVLYRIRFY FPRWYCSGSNRAYRHGISRGAEAPLLDDFVMSYLFAQWRHDFVHGWIKVPVTHETQEE C LGMAVLDMMRIAI<ENDQTPLAIYNSISYI<TFLPI<CIRAI<IQDYHILTRI<RIRYRFRRFIQ QFSQCKATARNLKLKYLINLETLQSAFYTEKFEVKEPGSGPSGEEIFATIUTGNGGIQWSR GKHKESETLTEQDLQLYCDFPNIIDVSIKQANQEGSNESRVVTIHKQDGKNLEIELSSLRE ALSFVSLIDGYYRLTADAHHYLCKEVAPPAVLENIQSNCHGPISMDFAISKLKKAGNQT

[0404] GLYVLRCSPKDFNKYFLTFAVERENVIEYKHCLITKNENEEYNLSGTKKNFSSLKDLLNC YQMETVRSDNIIFQFTKCCPPKPKDKSNLLVFRTNGVSDVPTSPTLQRPTHMNQMVFHKI RNEDLIFNESLGQGTFTKIFKGVRREVGDYGQLHETEVLLKVLDKAHRNYSESFFEAAS MMSKLSHKHLVLNYGVCVCGDENILVQEFVKFGSLDTYLKKNKNCINIL WKLEV AKQL AWAMHFLEENTLIHGNVCAKNILLIREEDRKTGNPPFIKLSDPGISITVLPKDILQERIPWV PPECIENPKNLNLATDKWSFGTTLWEICSGGDKPLSALDSQRKLQFYEDRHQLPAPKWA ELANLINNCMDYEPDFRPSFRAIIRDLNSLFTPDYELLTENDMLPNMRIGALGFSGAFED RDPTQFEERHLKFLQQLGKGNFGSVEMCRYDPLQDNTGEVVAVKKLQHSTEEHLRDFE REIEILKSLQHDNIVKYKGVCYSAGRRNLKLIMEYLPYGSLRDYLQKHKERIDHIKLLQY TSQICKGMEYLGTKRYIHRDLATRNILVENENRVKIGDFGLTKVLPQDKEYYKVKEPGE SPIFWYAPESLTESKFSVASDVWSFGVVLYELFTYIEKSKSPPAEFMRMIGNDKQGQMIV FHLIELLKNNGRLPRPDGCPDEIYMIMTECWNNNVNQRPSFRDLALRVDQIRDNMAG SEQ ID NO: 122. JAK3

[0405] MAPPSEETPLIPQRSCSLLSTEAGALHVLLPARGPGPPQRLSFSFGDHLAEDLCVQAAKA SGILPVYHSLFALATEDLSCWFPPSHIFSVEDASTQVLLYRIRFYFPNWFGLEKCHRFGLR KDLASAILDLPVLEHLFAQHRSDLVSGRLPVGLSLKEQGECLSLAVLDLARMAREQAQR PGELLKTVSYKACLPPSLRDLIQGLSFVTRRRIRRTVRRALRRVAACQADRHSLMAKYI MDLERLDPAGAAETFHVGLPGALGGHDGLGLLRVAGDGGIAWTQGEQEVLQPFCDFPE IVDISIKQAPRVGPAGEHRLVTVTRTDNQILEAEFPGLPEALSFVALVDGYFRLTTDSQHF FCKEVAPPRLLEEVAEQCHGPITLDFAINKLKTGGSRPGSYVLRRSPQDFDSFLLTVCVQ NPLGPDYKGCLIRRSPTGTFLLVGLSRPHSSLRELLATCWDGGLHVDGVAVTLTSCCIPR

[0406] PKEKSNLIVVQRGHSPPTSSLVQPQSQYQLSQMTFHKIPADSLEWHENLGHGSFTKIYRG CRHEVVDGEARKTEVLLKVMDAKHKNCMESFLEAASLMSQVSYRHLVLLHGVCMAG DSTMVQEFVHLGAIDMYLRKRGHLVPASWKLQVVKQLAYALNYLEDKGLPHGNVSAR KVLLAREGADGSPPFIKLSDPGVSPAVLSLEMLTDRIPWVAPECLREAQTLSLEADKWG FGATVWEVFSGVTMPISALDPAKKLQFYEDRQQLPAPKWTELALLIQQCMAYEPVQRP SFRAVIRDLNSLISSDYELLSDPTPGALAPRDGLWNGAQLYACQDPTIFEERHLKYISQLG

[0407] KGNFGSVELCRYDPLGDNTGALVAVKQLQHSGPDQQRDFQREIQILKALHSDFIVKYRG VSYGPGRQSLRLVMEYLPSGCLRDFLQRHRARLDASRLLLYSSQICKGMEYLGSRRCVH RDLAARNILVESEAHVKIADFGLAKLLPLDKDYYVVREPGQSPIFWYAPESLSDNIFSRQ SDVWSFGVVLYELFTYCDKSCSPSAEFLRMMGCERDVPALCRLLELLEEGQRLPAPPAC PAEVHELMKLCWAPSPQDRPSFSALGPQLDMLWSGSRGCETHAFTAHPEGKHHSLSFS SEQ ID NO: 123. Tyk2

[0408] MPLRHWGMARGSKP VGDGAQPMAAMGGLKVLLHWAGPGGGEPWVTF SE S SLT AEE V CIHIAHKVGITPPCFNLFALFDAQAQVWLPPNHILEIPRDASLMLYFRIRFYFRNWHGMN PREPAVYRCGPPGTEASSDQTAQGMQLLDPASFEYLFEQGKHEFVNDVASLWELSTEEE IHHFKNESLGMAFLHLCHLALRHGIPLEEVAKKTSFKDCIPRSFRRHIRQHSALTRLRLRN VFRRFLRDFQPGRLSQQMVMVKYLATLERLAPRFGTERVPVCHLRLLAQAEGEPCYIRD SGVAPTDPGPESAAGPPTHEVLVTGTGGIQWWPVEEEVNKEEGSSGSSGRNPQASLFGK KAKAHKAVGQPADRPREPLWAYFCDFRDITHVVLKEHCVSIHRQDNKCLELSLPSRAA ALSFVSLVDGYFRLTADSSHYLCHEVAPPRLVMSIRDGIHGPLLEPFVQAKLRPEDGLYL IHWSTSHPYRLILTVAQRSQAPDGMQSLRLRKFPIEQQDGAFVLEGWGRSFPSVRELGA ALQGCLLRAGDDCFSLRRCCLPQPGETSNLIIMRGARASPRTLNLSQLSFHRVDQKEITQ LSHLGQGTRTNVYEGRLRVEGSGDPEEGKMDDEDPLVPGRDRGQELRVVLKVLDPSHH DIALAFYETASLMSQVSHTHLAFVHGVCVRGPENIMVTEYVEHGPLDVWLRRERGHVP MAWKMVVAQQL AS AL S YLENKNLVHGNVC GRNILL A RLGLAEGT SPFIKLSDPGVGLG ALSREERVERIPWLAPECLPGGANSLSTAMDKWGFGATLLEICFDGEAPLQSRSPSEKEH F YQRQHRLPEP SCPQLATLT SQCLT YEPTQRP SFRTILRDLTRLQPHNL AD VLT VNPD SPA SDPTVFHKRYLKKIRDLGEGHFGKVSLYCYDPTNDGTGEMVAVKALKADCGPQHRSG WKQEIDILRTLYHEHIIKYKGCCEDQGEKSLQLVMEYVPLGSLRDYLPRHSIGLAQLLLF AQQICEGMAYLHAQHYIHRDLAARNVLLDNDRLVKIGDFGLAKAVPEGHEYYRVRED GDSPVFWYAPECLKEYKFYYASDVWSFGVTLYELLTHCDSSQSPPTKFLELIGIAQGQM TVLRLTELLERGERLPRPDKCPCEVYHLMKNCWETEASFRPTFENLIPILKTVHEKYQGQ

[0409] APSVFSVC

[0410] SEQ ID NO: 124. STAT1

[0411] MSQWYELQQLDSKFLEQVHQLYDDSFPMEIRQYLAQWLEKQDWEHAANDVSFATIRF HDLLSQLDDQYSRFSLENNFLLQHNIRKSKRNLQDNFQEDPIQMSMIIYSCLKEERKILEN AQRFNQAQSGNIQSTVMLDKQKELDSKVRNVKDKVMCIEHEIKSLEDLQDEYDFKCKT LQNREHETNGVAKSDQKQEQLLLKKMYLMLDNKRKEVVHKIIELLNVTELTQNALIND ELVEWKRRQQSACIGGPPNACLDQLQNWFTIVAESLQQVRQQLKKLEELEQKYTYEHD PITKNKQVLWDRTFSLFQQLIQSSFVVERQPCMPTHPQRPLVLKTGVQFTVKLRLLVKL QELNYNLKVKVLFDKDVNERNTVKGFRKFNILGTHTKVMNMEESTNGSLAAEFRHLQL KEQKNAGTRTNEGPLIVTEELHSLSFETQLCQPGLVIDLETTSLPVVVISNVSQLPSGWAS ILWYNMLVAEPRNLSFFLTPPCARWAQLSEVLSWQFSSVTKRGLNVDQLNMLGEKLLG PNASPDGLIPWTRFCKENINDKNFPFWLWIESILELIKKHLLPLWNDGCIMGFISKERERA LLKDQQPGTFLLRFSESSREGAITFTWVERSQNGGEPDFHAVEPYTKKELSAVTFPDIIRN YKVMAAENIPENPLKYLYPNIDKDHAFGKYYSRPKEAPEPMELDGPKGTGYIKTELISVS

[0412] EVHPSRLQTTDNLLPMSPEEFDEVSRIVGSVEFDSMMNTV

[0413] SEQ ID NO: 125. STAT2

[0414] MAQWEMLQNLDSPFQDQLHQLYSHSLLPVDIRQYLAVWIEDQNWQEAALGSDDSKAT

[0415] MLFFHFLDQLNYECGRCSQDPESLLLQHNLRKFCRDIQPFSQDPTQLAEMIFNLLLEEKRI LIQAQRAQLEQGEPVLETPVESQQHEIESRILDLRAMMEKLVKSISQLKDQQDVFCFRYK IQAKGKTPSLDPHQTKEQKILQETLNELDKRRKEVLDASKALLGRLTTLIELLLPKLEEW KAQQQKACIRAPIDHGLEQLETWFTAGAKLLFHLRQLLKELKGLSCLVSYQDDPLTKGV DLRNAQVTELLQRLLHRAFVVETQPCMPQTPHRPLILKTGSKFTVRTRLLVRLQEGNES LTVEVSIDRNPPQLQGFRKFNILTSNQKTLTPEKGQSQGLIWDFGYLTLVEQRSGGSGKG SNKGPLGVTEELHIISFTVKYTYQGLKQELKTDTLPVVIISNMNQLSIAWASVLWFNLLSP NLQNQQFFSNPPKAPWSLLGPALSWQFSSYVGRGLNSDQLSMLRNKLFGQNCRTEDPL LSWADFTKRESPPGKLPFWTWLDKILELVHDHLKDLWNDGRIMGFVSRSQERRLLKKT

[0416] MSGTFLLRFSESSEGGITCSWVEHQDDDKVLIYSVQPYTKEVLQSLPLTEIIRHYQLLTEE NIPENPLRFLYPRIPRDEAFGCYYQEKVNLQERRKYLKHRLIVVSNRQVDELQQPLELKP EPELESLELELGLVPEPELSLDLEPLLKAGLDLGPELESVLESTLEPVIEPTLCMVSQTVPE PDQGPVSQPVPEPDLPCDLRHLNTEPMEIFRNCVKIEEIMPNGDPLLAGQNTVDEVYVSR P SHF YTDGPLMP SDF

[0417] SEQ ID NO: 126. STAT3

[0418] MAQWNQLQQLDTRYLEQLHQLYSDSFPMELRQFLAPWIESQDWAYAASKESHATLVF HNLLGEIDQQYSRFLQESNVLYQHNLRRIKQFLQSRYLEKPMEIARIVARCLWEESRLLQ TAATAAQQGGQANHPTAAVVTEKQQMLEQHLQDVRKRVQDLEQKMKVVENLQDDFD

[0419] FNYKTLKSQGDMQDLNGNNQSVTRQKMQQLEQMLTALDQMRRSIVSELAGLLSAMEY VQKTLTDEELADWKRRQQIACIGGPPNICLDRLENWITSLAESQLQTRQQIKKLEELQQK VSYKGDPIVQHRPMLEERIVELFRNLMKSAFVVERQPCMPMHPDRPLVIKTGVQFTTKV RLLVKFPELNYQLKIKVCIDKDSGDVAALRGSRKFNILGTNTKVMNMEESNNGSLSAEF KHLTLREQRCGNGGRANCDASLIVTEELHLITFETEVYHQGLKIDLETHSLPVVVISNICQ MPNAWASILWYNMLTNNPKNVNFFTKPPIGTWDQVAEVLSWQFSSTTKRGLSIEQLTTL AEKLLGPGVNYSGCQITWAKFCKENMAGKGFSFWVWLDNIIDLVKKYILALWNEGYIM GFISKERERAILSTKPPGTFLLRFSESSKEGGVTFTWVEKDISGKTQIQSVEPYTKQQLNN MSFAEIIMGYKIMDATNILVSPLVYLYPDIPKEEAFGKYCRPESQEHPEADPGSAAPYLK TKFICVTPTTCSNTIDLPMSPRTLDSLMQFGNNGEGAEPSAGGQFESLTFDMELTSECATS PM

[0420] SEQ ID NO: 127. STAT4

[0421] MSQWNQVQQLEIKFLEQVDQFYDDNFPMEIRHLLAQWIENQDWEAASNNETMATILLQ NLLIQLDEQLGRVSKEKNLLLIHNLKRIRKVLQGKFHGNPMHVAVVISNCLREERRILAA ANMPVQGPLEKSLQSSSVSERQRNVEHKVAAIKNSVQMTEQDTKYLEDLQDEFDYRYK TIQTMDQSDKNSAMVNQEVLTLQEMLNSLDFKRKEALSKMTQIIHETDLLMNTMLIEEL QDWKRRQQIACIGGPLHNGLDQLQNCFTLLAESLFQLRRQLEKLEEQSTKMTYEGDPIP MQRTHMLERVTFLIYNLFKNSFVVERQPCMPTHPQRPLVLKTLIQFTVKLRLLIKLPELN YQVKVKASIDKNVSTLSNRRFVLCGTNVKAMSIEESSNGSLSVEFRHLQPKEMKSSAGG KGNEGCHMVTEELHSITFETQICLYGLTIDLETSSLPVVMISNVSQLPNAWASIIWYNVST NDSQNLVFFNNPPPATLSQLLEVMSWQFSSYVGRGLNSDQLHMLAEKLTVQSSYSDGH LTWAKFCKEHLPGKSFTFWTWLEAILDLIKKHILPLWIDGYVMGFVSKEKERLLLKDKM

[0422] PGTFLLRFSESHLGGITFTWVDHSESGEVRFHSVEPYNKGRLSALPFADILRDYKVIMAE NIPENPLKYLYPDIPKDKAFGKHYSSQPCEVSRPTERGDKGYVPSVFIPISTIRSDSTEPHS P SDLLPMSP S VYAVLRENL SPTTIET AMKSP YS AE

[0423] SEQ ID NO: 128. STAT5A

[0424] MAGWIQAQQLQGDALRQMQVLYGQHFPIEVRHYLAQWIESQPWDAIDLDNPQDRAQA TQLLEGLVQELQKKAEHQVGEDGFLLKIKLGHYATQLQKTYDRCPLELVRCIRHILYNE QRLVREANNCSSPAGILVDAMSQKHLQINQTFEELRLVTQDTENELKKLQQTQEYFIIQY QESLRIQAQFAQLAQLSPQERLSRETALQQKQVSLEAWLQREAQTLQQYRVELAEKHQ KTLQLLRKQQTIILDDELIQWKRRQQLAGNGGPPEGSLDVLQSWCEKLAEIIWQNRQQIR RAEHLCQQLPIPGPVEEMLAEVNATITDIISALVTSTFIIEKQPPQVLKTQTKFAATVRLLV GGKLNVHMNPPQVKATIISEQQAKSLLKNENTRNECSGEILNNCCVMEYHQATGTLSAH FRNMSLKRIKRADRRGAESVTEEKFTVLFESQFSVGSNELVFQVKTLSLPVVVIVHGSQD HNATATVLWDNAFAEPGRVPFAVPDKVLWPQLCEALNMKFKAEVQSNRGLTKENLVF LAQKLFNNS S SHLED YSGLS VSWSQFNRENLPGWNYTFWQWFDGVMEVLKKHHKPH WNDGAILGFVNKQQAHDLLINKPDGTFLLRFSDSEIGGITIAWKFDSPERNLWNLKPFTT RDF SIRSLADRLGDL S YLIYVFPDRPKDE VF SKYYTP VL AKAVDGYVKPQIKQ VVPEF VN AS AD AGGS S ATYMDQ AP SP AVCPQ AP YNMYPQNPDHVLDQDGEFDLDETMD VARHVE ELLRRPMD SLD SRL SPP AGLFT S ARGSL S SEQ ID NO: 129. STAT5B

[0425] MAVWIQAQQLQGEALHQMQALYGQHFPIEVRHYLSQWIESQAWDSVDLDNPQENIKA TQLLEGLVQELQKKAEHQVGEDGFLLKIKLGHYATQLQNTYDRCPMELVRCIRHILYNE QRLVREANNGSSPAGSLADAMSQKHLQINQTFEELRLVTQDTENELKKLQQTQEYFIIQ YQESLRIQAQFGPLAQLSPQERLSRETALQQKQVSLEAWLQREAQTLQQYRVELAEKHQ KTLQLLRKQQTIILDDELIQWKRRQQLAGNGGPPEGSLDVLQSWCEKLAEIIWQNRQQIR RAEHLCQQLPIPGPVEEMLAEVNATITDIISALVTSTFIIEKQPPQVLKTQTKFAATVRLLV GGKLNVHMNPPQVKATIISEQQAKSLLKNENTRNDYSGEILNNCCVMEYHQATGTLSA HFRNMSLKRIKRSDRRGAESVTEEKFTILFESQFSVGGNELVFQVKTLSLPVWIVHGSQ DNNATATVLWDNAFAEPGRVPFAVPDKVLWPQLCEALNMKFKAEVQSNRGLTKENLV FLAQKLFNNS S SHLEDYSGLS VSWSQFNRENLPGRNYTFWQWFDGVMEVLKKHLKPH WNDGAILGFVNKQQAHDLLINKPDGTFLLRFSDSEIGGITIAWKFDSQERMFWNLMPFT TRDFSIRSLADRLGDLNYLIYVFPDRPKDEVYSKYYTPVPCESATAKAVDGYVKPQIKQ VVPEF VNAS AD AGGGS ATYMDQ AP SP A VCPQ AHYNMYPQNPD S VLDTDGDFDLEDTM DVARRVEELLGRPMDSQWIPHAQS

[0426] Motility signaling

[0427] SEQ ID NO: 130. Rafi

[0428] MEHIQGAWKTISNGFGFKDAVFDGSSCISPTIVQQFGYQRRASDDGKLTDPSKTSNTIRV FLPNKQRTVVNVRNGMSLHDCLMKALKVRGLQPECCAVFRLLHEHKGKKARLDWNT DAASLIGEELQVDFLDHVPLTTHNFARKTFLKLAFCDICQKFLLNGFRCQTCGYKFHEH CSTKVPTMCVDWSNIRQLLLFPNSTIGDSGVPALPSLTMRRMRESVSRMPVSSQHRYSTP HAFTFNTSSPSSEGSLSQRQRSTSTPNVHMVSTTLPVDSRMIEDAIRSHSESASPSALSSSP NNLSPTGWSQPKTPVPAQRERAPVSGTQEKNKIRPRGQRDSSYYWEIEASEVMLSTRIGS GSFGTVYKGKWHGDVAVKILKVVDPTPEQFQAFRNEVAVLRKTRHVNILLFMGYMTK DNLAIVTQWCEGSSLYKHLHVQETKFQMFQLIDIARQTAQGMDYLHAKNIIHRDMKSN NIFLHEGLTVKIGDFGLATVKSRWSGSQQVEQPTGSVLWMAPEVIRMQDNNPFSFQSDV YSYGIVLYELMTGELPYSHINNRDQIIFMVGRGYASPDLSKLYKNCPKAMKRLVADCVK KVKEERPLFPQILSSIELLQHSLPKINRSASEPSLHRAAHTEDINACTLTTSPRLPVF SEQ ID NO: 131. cAMP MKTQRDGHSLGRWSLVLLLLGLVMPLAIIAQVLSYKEAVLRAIDGINQRSSDANLYRLL DLDPRPTMDGDPDTPKPVSFTVKETVCPRTTQQSPEDCDFKKDGLVKRCMGTVTLNQA RGSFDISCDKDNKRFALLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES

[0429] SEQ ID NO: 132. cdc42

[0430] MQTIKCVVVGDGAVGKTCLLISYTTNKFPSEYVPTVFDNYAVTVMIGGEPYTLGLFDTA GQEDYDRLRPLSYPQTDVFLVCFSVVSPSSFENVKEKWVPEITHHCPKTPFLLVGTQIDL RDDPSTIEKLAKNKQKPITPETAEKLARDLKAVKYVECSALTQKGLKNVFDEAILAALEP PEPKKSRRCVLL

[0431] SEQ ID NO: 133. Slug

[0432] MPRSFLVKKHFNASKKPNYSELDTHTVIISPYLYESYSMPVIPQPEILSSGAYSPITVWTT AAPFHAQLPNGLSPLSGYSSSLGRVSPPPPSDTSSKDHSGSESPISDEEERLQSKLSDPHAI EAEKFQCNLCNKTYSTFSGLAKHKQLHCDAQSRKSFSCKYCDKEYVSLGALKMHIRTH TLPCVCKICGKAFSRPWLLQGHIRTHTGEKPFSCPHCNRAFADRSNLRAHLQTHSDVKK YQCKNC SKTF SRMSLLHKHEESGCC VAH

[0433] SEQ ID NO: 134. Snail

[0434] MPRSFL VRKP SDPNRKPNYSELQD SNPEFTFQQP YDQ AHLL AAIPPPEILNPT ASLPMLIW DSVLAPQAQPIAWASLRLQESPRVAELTSLSDEDSGKGSQPPSPPSPAPSSFSSTSVSSLEA EAYAAFPGLGQVPKQLAQLSEAKDLQARKAFNCKYCNKEYLSLGALKMHIRSHTLPCV CGTCGKAFSRPWLLQGHVRTHTGEKPFSCPHCSRAFADRSNLRAHLQTHSDVKKYQCQ ACARTF SRMSLLHKHQESGC SGCPR

[0435] SEQ ID NO: 135. Twist

[0436] MMQDVSSSPVSPADDSLSNSEEEPDRQQPPSGKRGGRKRRSSRRSAGGGAGPGGAAGG GVGGGDEPGSPAQGKRGKKSAGCGGGGGAGGGGGSSSGGGSPQSYEELQTQRVMANV RERQRTQ SLNEAF AALRKIIPTLP SDKLSKIQTLKLAARYIDFL YQ VLQ SDELD SKMASC S YVAHERLS YAF S VWRMEGAWSMS ASH

[0437] SEQ ID NO: 136. MAPK11

[0438] MSGPRAGFYRQELNKTVWEVPQRLQGLRPVGSGAYGSVCSAYDARLRQKVAVKKLSR PFQSLIHARRTYRELRLLKHLKHENVIGLLDVFTPATSIEDFSEVYLVTTLMGADLNNIVK CQALSDEHVQFLVYQLLRGLKYIHSAGIIHRDLKPSNVAVNEDCELRILDFGLARQADEE MTGYVATRWYRAPEIMLNWMHYNQTVDIWSVGCIMAELLQGKALFPGSDYIDQLKRI MEVVGTP SPEVLAKIS SEHART YIQ SLPPMPQKDL S SIFRGANPL AIDLLGRMLVLD SDQ RVSAAEALAHAYFSQYHDPEDEPEAEPYDESVEAKERTLEEWKELTYQEVLSFKPPEPP KPPGSLEIEQ

[0439] SEQ ID NO: 137. MAPK12

[0440] MSSPPPARSGFYRQEVTKTAWEVRAVYRDLQPVGSGAYGAVCSAVDGRTGAKVAIKK LYRPFQSELFAKRAYRELRLLKHMRHENVIGLLDVFTPDETLDDFTDFYLVMPFMGTDL GKLMKHEKLGEDRIQFLVYQMLKGLRYIHAAGIIHRDLKPGNLAVNEDCELKILDFGLA RQADSEMTGYVVTRWYRAPEVILNWMRYTQTVDIWSVGCIMAEMITGKTLFKGSDHL DQLKEIMKVTGTPPAEFVQRLQSDEAKNYMKGLPELEKKDFASILTNASPLAVNLLEKM

[0441] LVLDAEQRVTAGEALAHPYFESLHDTEDEPQVQKYDDSFDDVDRTLDEWKRVTYKEV LSFKPPRQLGARVSKETPL

[0442] SEQ ID NO: 138. MAPK13

[0443] MSLIRKKGFYKQDVNKTAWELPKTYVSPTHVGSGAYGSVCSAIDKRSGEKVAIKKLSRP FQSEIFAKRAYRELLLLKHMQHENVIGLLDVFTPASSLRNFYDFYLVMPFMQTDLQKIM GMEFSEEKIQYLVYQMLKGLKYIHSAGVVHRDLKPGNLAVNEDCELKILDFGLARHAD AEMTGYVVTRWYRAPEVILSWMHYNQTVDIWSVGCIMAEMLTGKTLFKGKDYLDQLT QILKVTGVPGTEFVQKLNDKAAKSYIQSLPQTPRKDFTQLFPRASPQAADLLEKMLELD

[0444] VDKRLTAAQALTHPFFEPFRDPEEETEAQQPFDDSLEHEKLTVDEWKQHIYKEIVNFSPI ARKDSRRRSGMKL

[0445] SEQ ID NO: 139. MAPK14

[0446] MSQERPTFYRQELNKTIWEVPERYQNLSPVGSGAYGSVCAAFDTKTGLRVAVKKLSRPF QSIIHAKRTYRELRLLKHMKHENVIGLLDVFTPARSLEEFNDVYLVTHLMGADLNNIVK CQKLTDDHVQFLIYQILRGLKYIHSADIIHRDLKPSNLAVNEDCELKILDFGLARHTDDE MTGYVATRWYRAPEIMLNWMHYNQTVDIWSVGCIMAELLTGRTLFPGTDHIDQLKLIL RLVGTPGAELLKKISSESARNYIQSLTQMPKMNFANVFIGANPLAVDLLEKMLVLDSDK

[0447] RITAAQALAHAYFAQYHDPDDEPVADPYDQSFESRDLLIDEWKSLTYDEVISFVPPPLDQ EEMES

[0448] SEQ ID NO: 140. c-jun

[0449] MTAKMETTFYDDALNASFLPSESGPYGYSNPKILKQSMTLNLADPVGSLKPHLRAKNSD LLTSPDVGLLKLASPELERLIIQSSNGHITTTPTPTQFLCPKNVTDEQEGFAEGFVRALAEL HSQNTLPSVTSAAQPVNGAGMVAPAVASVAGGSGSGGFSASLHSEPPVYANLSNFNPG ALSSGGGAPSYGAAGLAFPAQPQQQQQPPHHLPQQMPVQHPRLQALKEEPQTVPEMPG ETPPLSPIDMESQERIKAERKRMRNRIAASKCRKRKLERIARLEEKVKTLKAQNSELAST ANMLREQVAQLKQKVMNHVNSGCQLMLTQQLQTF SEQ ID NO: 141. c-fos

[0450] MMFSGFNADYEASSSRCSSASPAGDSLSYYHSPADSFSSMGSPVNAQDFCTDLAVSSAN FIPTVTAISTSPDLQWLVQPALVSSVAPSQTRAPHPFGVPAPSAGAYSRAGVVKTMTGGR AQSIGRRGKVEQLSPEEEEKRRIRRERNKMAAAKCRNRRRELTDTLQAETDQLEDEKSA LQTEIANLLKEKEKLEFILAAHRPACKIPDDLGFPEEMSVASLDLTGGLPEVATPESEEAF TLPLLNDPEPKPSVEPVKSISSMELKTEPFDDFLFPASSRPSGSETARSVPDMDLSGSFYA ADWEPLHSGSLGMGPMATELEPLCTPVVTCTPSCTAYTSSFVFTYPEADSFPSCAAAHR KGSSSNEPSSDSLSSPTLLAL SEQ ID NO: 142. c-Myc

[0451] MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAPSEDIWKKFELLPTP PLSP SRRSGLC SP S YVAVTPF SLRGDNDGGGGSF ST ADQLEMVTELLGGDMVNQ SFICDP DDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPNPARGHSVCSTSSLYLQ DLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDSLLSSTESSPQGSPEPLVLHE ETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGSPSAGGHSKPPHSPLVLKRCHVS THQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNRKCTSPRSSDTEENVKRRTHNVL ERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKATAYILSVQAEEQKLISEEDLLRKR REQLKHKLEQLRNSCA SEQ ID NO: 143. Pim-1

[0452] MLLSKINSLAHLRAAPCNDLHATKLAPGKEKEPLESQYQVGPLLGSGGFGSVYSGIRVS DNLPVAIKHVEKDRISDWGELPNGTRVPMEVVLLKKVSSGFSGVIRLLDWFERPDSFVLI LERPEPVQDLFDFITERGALQEELARSFFWQVLEAVRHCHNCGVLHRDIKDENILIDLNR GELKLIDFGSGALLKDTVYTDFDGTRVYSPPEWIRYHRYHGRSAAVWSLGILLYDMVC GDIPFEHDEEIIRGQ VFFRQRVS SECQHLIRWCL ALRP SDRPTFEEIQNHP WMQD VLLPQE TAEIHLHSLSPGPSK SEQ ID NO: 144. Gsk3b

[0453] MSGRPRTTSFAESCKPVQQPSAFGSMKVSRDKDGSKVTTVVATPGQGPDRPQEVSYTD TKVIGNGSFGVVYQAKLCDSGELVAIKKVLQDKRFKNRELQIMRKLDHCNIVRLRYFFY SSGEKKDEVYLNLVLDYVPETVYRVARHYSRAKQTLPVIYVKLYMYQLFRSLAYIHSFG ICHRDIKPQNLLLDPDTAVLKLCDFGS AKQL VRGEPNVS YIC SRYYRAPELIFGATD YTS S IDVWSAGCVLAELLLGQPIFPGDSGVDQLVEIIKVLGTPTREQIREMNPNYTEFKFPQIKA HPWTKVFRPRTPPEAIALCSRLLEYTPTARLTPLEACAHSFFDELRDPNVKLPNGRDTPA LFNFTTQELSSNPPLATILIPPHARIQAAASTPTNATAASDANTGDRGQTNNAASASASNS T

[0454] SEQ ID NO: 145. Akt

[0455] MSDVAIVKEGWLHKRGEYIKTWRPRYFLLKNDGTFIGYKERPQDVDQREAPLNNFSVA QCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWTTAIQTVADGLKKQEEEEM DFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEFEYLKLLGKGTFGKVILVKEKATGRYY AMKILKKEVIVAKDEVAHTLTENRVLQNSRHPFLTALKYSFQTHDRLCFVMEYANGGE LFFHLSRERVFSEDRARFYGAEIVSALDYLHSEKNVVYRDLKLENLMLDKDGHIKITDF GLCKEGIKDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMYEMMCGRLPF YNQDHEKLFELILMEEIRFPRTLGPEAKSLLSGLLKKDPKQRLGGGSEDAKEIMQHRFFA GIVWQHVYEKKLSPPFKPQVTSETDTRYFDEEFTAQMITITPPDQDDSMECVDSERRPHF PQFSYSASGTA

[0456] SEQ ID NO: 146. MCK

[0457] MPFGNTHNKFKLNYKPEEEYPDLSKHNNHMAKVLTLELYKKLRDKETPSGFTVDDVIQ TGVDNPGHPFIMTVGCVAGDEESYEVFKELFDPIISDRHGGYKPTDKHKTDLNHENLKG GDDLDPNYVL S SRVRTGRSIKGYTLPPHC SRGERRAVEKLS VE ALNSLTGEFKGK YYPL KSMTEKEQQQLIDDHFLFDKPVSPLLLASGMARDWPDARGIWHNDNKSFLVWVNEED HLRVISMEKGGNMKEVFRRFCVGLQKIEEIFKKAGHPFMWNQHLGYVLTCPSNLGTGL RGGVHVKL AHL SKHPKFEEILTRLRLQKRGTGGVDTAA VGS VFD VSNADRLGS SEVEQ VQLVVDGVKLMVEMEKKLEKGQSIDDMIPAQK

[0458] SEQ ID NO: 147. IRF1

[0459] MPITRMRMRPWLEMQINSNQIPGLIWINKEEMIFQIPWKHAAKHGWDINKDACLFRSW AIHTGRYK AGEKEPDPKTWK ANFRC AMNSLPDIEEVKDQ SRNKGS S AVRVYRMLPPLT KNQRKERKSKSSRDAKSKAKRKSCGDSSPDTFSDGLSSSTLPDDHSSYTVPGYMQDLEV EQ AL TP AL SPC AVS STLPDWHIP VEVVPD STSDL YNFQ VSPMP ST SEATTDEDEEGKLPE DIMKLLEQSEWQPTNVDGKGYLLNEPGVQPTSVYGDFSCKEEPEIDSPGGDIGLSLQRVF TDLKNMD ATWLD SLLTP VRLP SIQ AIPC AP

[0460] SEQ ID NO: 148. TRADD MAAGQNGHEEWVGSAYLFVESSLDKVVLSDAYAHPQQKVAVYRALQAALAESGGSP DVLQMLKIHRSDPQLIVQLRFCGRQPCGRFLRAYREGALRAALQRSLAAALAQHSVPLQ LELRAGAERLDALLADEERCLSCILAQQPDRLRDEELAELEDALRNLKCGSGARGGDGE VASAPLQPPVPSLSEVKPPPPPPPAQTFLFQGQPVVNRPLSLKDQQTFARSVGLKWRKVG RSLQRGCRALRDPALDSLAYEYEREGLYEQAFQLLRRFVQAEGRRATLQRLVEALEENE LT SLAEDLLGLTDPNGGL A

[0461] SEQ ID NO: 149. TRAF2

[0462] MAAASVTPPGSLELLQPGFSKTLLGTKLEAKYLCSACRNVLRRPFQAQCGHRYCSFCLA SILSSGPQNCAACVHEGIYEEGISILESSSAFPDNAARREVESLPAVCPSDGCTWKGTLKE YESCHEGRCPLMLTECPACKGLVRLGEKERHLEHECPERSLSCRHCRAPCCGADVKAH

[0463] HEVCPKFPLTCDGCGKKKIPREKFQDHVKTCGKCRVPCRFHAIGCLETVEGEKQQEHEV QWLREHLAMLLSSVLEAKPLLGDQSHAGSELLQRCESLEKKTATFENIVCVLNREVERV AMTAEAC SRQHRLDQDKIEALS SKVQQLERSIGLKDLAMADLEQKVLEMEASTYDGVF IWKISDFARKRQEAV AGRIP AIFSPAFYTSRYGYKMCLRIYLNGDGTGRGTHLSLFFVVM KGPNDALLRWPFNQKVTLMLLDQNNREHVIDAFRPDVTSSSFQRPVNDMNIASGCPLFC PVSKMEAKNSYVRDDAIFIKAIVDLTGL

[0464] SEQ ID NO: 150. FADD

[0465] MDPFLVLLHS VS S SLS S SELTELKFLCLGRVGKRKLERVQSGLDLF SMLLEQNDLEPGHT ELLRELLASLRRHDLLRRVDDFEAGAAAGAAPGEEDLCAAFNVICDNVGKDWRRLARQ LKVSDTKIDSIEDRYPRNLTERVRESLRIWKNTEKENATVAHLVGALRSCQMNLVADLV QEVQQARDLQNRSGAMSPMSWNSDASTSEAS

[0466] SEQ ID NO: 151. RIPK1

[0467] MQPDMSLNVIKMKSSDFLESAELDSGGFGKVSLCFHRTQGLMIMKTVYKGPNCIEHNE ALLEEAKMMNRLRHSRVVKLLGVIIEEGKYSLVMEYMEKGNLMHVLKAEMSTPLSVK GRIILEIIEGMCYLHGKGVIHKDLKPENILVDNDFHIKIADLGLASFKMWSKLNNEEHNEL

[0468] REVDGTAKKNGGTLYYMAPEHLNDVNAKPTEKSDVYSFAVVLWAIFANKEPYENAICE QQLIMCIKSGNRPDVDDITEYCPREIISLMKLCWEANPEARPTFPGIEEKFRPFYLSQLEES VEEDVKSLKKEYSNENAVVKRMQSLQLDCVAVPSSRSNSATEQPGSLHSSQGLGMGPV EESWFAPSLEHPQEENEPSLQSKLQDEANYHLYGSRMDRQTKQQPRQNVAYNREEERR RRVSHDPFAQQRPYENFQNTEGKGTAYSSAASHGNAVHQPSGLTSQPQVLYQNNGLYS SHGFGTRPLDPGTAGPRVWYRPIPSHMPSLHNIPVPETNYLGNTPTMPFSSLPPTDESIKY TIYNSTGIQIGAYNYMEIGGTSSSLLDSTNTNFKEEPAAKYQAIFDNTTSLTDKHLDPIRE NLGKHWKNCARKLGFTQSQIDEIDHDYERDGLKEKVYQMLQKWVMREGIKGATVGKL AQ ALHQC SRIDLL S SLI YVSQN SEQ ID NO: 152. ASK1

[0469] MSTEADEGITFSVPPFAPSGFCTIPEGGICRRGGAAAVGEGEEHQLPPPPPGSFWNVESAA APGIGCPAATS SS S ATRGRGS S VGGGSRRTTVAYVINEASQGQLVVAESEALQSLREACE TVGATLETLHFGKLDFGETTVLDRFYNADIAVVEMSDAFRQPSLFYHLGVRESFSMANN IILYCDTNSDSLQSLKEIICQKNTMCTGNYTFVPYMITPHNKVYCCDSSFMKGLTELMQP NFELLLGPICLPLVDRFIQLLKVAQASSSQYFRESILNDIRKARNLYTGKELAAELARIRQ RVDNIEVLTADIVINLLLSYRDIQDYDSIVKLVETLEKLPTFDLASHHHVKFHYAFALNR RNLPGDRAKALDIMIPMVQSEGQVASDMYCLVGRIYKDMFLDSNFTDTESRDHGASWF KK AFESEPTLQ SGINYAVLLLAAGHQFES SFELRKVGVKL S SLLGKKGNLEKLQ S YWE V GFFLGASVLANDHMRVIQASEKLFKLKTPAWYLKSIVETILIYKHFVKLTTEQPVAKQEL VDFWMDFLVEATKTDVTVVRFPVLILEPTKIYQPSYLSINNEVEEKTISIWHVLPDDKKGI HEWNFSASSVRGVSISKFEERCCFLYVLHNSDDFQIYFCTELHCKKFFEMVNTITEEKGR STEEGDCESDLLEYDYEYDENGDRVVLGKGTYGIVYAGRDLSNQVRIAIKEIPERDSRYS QPLHEEIALHKHLKHKNIVQYLGSFSENGFIKIFMEQVPGGSLSALLRSKWGPLKDNEQT IGFYTKQILEGLKYLHDNQIVHRDIKGDNVLINTYSGVLKISDFGTSKRLAGINPCTETFT GTLQYMAPEIIDKGPRGYGKAADIWSLGCTIIEMATGKPPFYELGEPQAAMFKVGMFKV HPEIPESMSAEAKAFILKCFEPDPDKRACANDLLVDEFLKVSSKKKKTQPKLSALSAGSN EYLRSISLPVPVLVEDTSSSSEYGSVSPDTELKVDPFSFKTRAKSCGERDVKGIRTLFLGIP DENFEDHSAPPSPEEKDSGFFMLRKDSERRATLHRILTEDQDKIVRNLMESLAQGAEEPK LKWEHITTLIASLREFVRSTDRKIIATTLSKLKLELDFDSHGISQVQVVLFGFQDAVNKVL RNHNIKPHWMFALDSIIRKAVQTAITILVPELRPHFSLASESDTADQEDLDVEDDHEEQPS NQTVRRPQAVIEDAVATSGVSTLSSTVSHDSQSAHRSLNVQLGRMKIETNRLLEELVRK EKELQALLHRAIEEKDQEIKHLKLKSQPIEIPELPVFHLNSSGTNTEDSELTDWLRVNGAD EDTISRFLAEDYTLLDVLYYVTRDDLKCLRLRGGMLCTLWKAIIDFRNKQT SEQ ID NO: 153. MEKK1

[0470] MAAAAGNRAS S SGFPGARATSPEAGGGGGALKAS S APAAAAGLLREAGSGGRERADW RRRQLRKVRSVELDQLPEQPLFLAASPPASSTSPSPEPADAAGSGTGFQPVAVPPPHGAA SRGGAHLTE S VAAPD SGAS SP AAAEPGEKRAP AAEP SP AA AP AGREMENKETLKGLHK MDDRPEERMIREKLKATCMPAWKHEWLERRNRRGPVVVKPIPVKGDGSEMNF1LAAES PGEVQASAASPASKGRRSPSPGNSPSGRTVKSESPGVRRKRVSPVPFQSGRITPPRRAPSP DGF SP YSPEETNRRVNKVMRARL YLLQQIGPNSFLIGGD SPDNKYRVFIGPQNC SC ARGT FCIHLLFVMLRVFQLEPSDPMLWRKTLKNFEVESLFQKYHSRRSSRIKAPSRNTIQKFVS RMSNSHTLSSSSTSTSSSENSIKDEEEQMCPICLLGMLDEESLTVCEDGCRNKLHHHCMSI WAEECRRNREPLICPLCRSKWRSHDFYSHELSSPVDSPSSLRAAQQQTVQQQPLAGSRR NQESNFNLTHYGTQQIPPAYKDLAEPWIQVFGMELVGCLFSRNWNVREMALRRLSHDV SGALLLANGESTGNSGGSSGSSPSGGATSGSSQTSISGDVVEACCSVLSMVCADPVYKV YVAALKTLRAMLVYTPCHSLAERIKLQRLLQPVVDTILVKCADANSRTSQLSISTLLELC KGQAGELAVGREILKAGSIGIGGVDYVLNCILGNQTESNNWQELLGRLCLIDRLLLEFPA EFYPHIVSTDVSQAEPVEIRYKKLLSLLTFALQSIDNSHSMVGKLSRRIYLSSARMVTTVP HVFSKLLEMLSVSSSTHFTRMRRRLMAIADEVEIAEAIQLGVEDTLDGQQDSFLQASVPN NYLETTENSSPECTVHLEKTGKGLCATKLSASSEDISERLASISVGPSSSTTTTTTTTEQPK PMVQTKGRPHSQCLNSSPLSHHSQLMFPALSTPSSSTPSVPAGTATDVSKHRLQGFIPCRI P S ASPQTQRKF SLQFHRNCPENKDSDKL SP VFTQ SRPLP S SNIHRPKP SRPTPGNTSKQGD PSKNSMTLDLNS S SKCDDSFGC S SNS SNAVIPSDETVFTPVEEKCRLDVNTELNS SIEDLL EASMPSSDTTVTFKSEVAVLSPEKAENDDTYKDDVNHNQKCKEKMEAEEEEALAIAMA MSASQDALPIVPQLQVENGEDIIIIQQDTPETLPGHTKAKQPYREDTEWLKGQQIGLGAFS SCYQAQDVGTGTLMAVKQVTYVRNTSSEQEEVVEALREEIRMMSHLNHPNIIRMLGAT CEKSNYNLFIEWMAGGSVAHLLSKYGAFKESVVINYTEQLLRGLSYLHENQIIHRDVKG ANLLIDSTGQRLRIADFGAAARLASKGTGAGEFQGQLLGTIAFMAPEVLRGQQYGRSCD VWSVGCAIIEMACAKPPWNAEKHSNHLALIFKIASATTAPSIPSHLSPGLRDVALRCLEL QPQDRPPSRELLKHPVFRTTW SEQ ID NO: 154. MKK7

[0471] MAASSLEQKLSRLEAKLKQENREARRRIDLNLDISPQRPRPTLQLPLANDGGSRSPSSESS PQHPTPPARPRHMLGLPSTLFTPRSMESIEIDQKLQEIMKQTGYLTIGGQRYQAEINDLEN LGEMGSGTCGQVWKMRFRKTGHVIAVKQMRRSGNKEENKRILMDLDVVLKSHDCPYI VQCFGTFITNTDVFIAMELMGTCAEKLKKRMQGPIPERILGKMTVAIVKALYYLKEKHG VIHRDVKPSNILLDERGQIKLCDFGISGRLVDSKAKTRSAGCAAYMAPERIDPPDPTKPD YDIRADVWSLGISLVELATGQFPYKNCKTDFEVLTKVLQEEPPLLPGHMGFSGDFQSFV KDCLTKDHRKRPKYNKLLEHSFIKRYETLEVDVASWFKDVMAKTESPRTSGVLSQPHLP FFR

[0472] SEQ ID NO: 155. JNK1

[0473] MSRSKRDNNFYSVEIGDSTFTVLKRYQNLKPIGSGAQGIVCAAYDAILERNVAIKKLSRP FQNQTHAKRAYRELVLMKCVNHKNIIGLLNVFTPQKSLEEFQDVYIVMELMDANLCQV IQMELDHERMSYLLYQMLCGIKHLHSAGIIHRDLKPSNIVVKSDCTLKILDFGLARTAGT SFMMTPYVVTRYYRAPEVILGMGYKENVDLWSVGCIMGEMVCHKILFPGRDYIDQWN KVIEQLGTPCPEFMKKLQPTVRTYVENRPKYAGYSFEKLFPDVLFPADSEHNKLKASQA RDLLSKMLVIDASKRISVDEALQHPYINVWYDPSEAEAPPPKIPDKQLDEREHTIEEWKE LIYKEVMDLEERTKNGVIRGQP SPLGAAVINGSQHP S S SS S VND VS SMSTDPTL ASDTD S SLEAAAGPLGCCR

[0474] SEQ ID NO: 156. NEMO

[0475] MNRHLWKSQLCEMVQPSGGPAADQDVLGEESPLGKPAMLHLPSEQGAPETLQRCLEEN QELRDAIRQSNQILRERCEELLHFQASQREEKEFLMCKFQEARKLVERLGLEKLDLKRQ

[0476] KEQALREVEHLKRCQQQMAEDKASVKAQVTSLLGELQESQSRLEAATKECQALEGRAR AASEQARQLESEREALQQQHSVQVDQLRMQGQSVEAALRMERQAASEEKRKLAQLQV AYHQLFQEYDNHIKSSVVGSERKRGMQLEDLKQQLQQAEEALVAKQEVIDKLKEEAEQ HKIVMETVPVLKAQADIYKADFQAERQAREKLAEKKELLQEQLEQLQREYSKLKASCQ ESARIEDMRKRHVEVSQAPLPPAPAYLSSPLALPSQRRSPPEEPPDFCCPKCQYQAPDMD TLQIHVMECIE

[0477] SEQ ID NO: 157. IKKA

[0478] MERPPGLRPGAGGPWEMRERLGTGGFGNVCLYQHRELDLKIAIKSCRLELSTKNRERW CHEIQIMKKLNHANVVKACDVPEELNILIHDVPLLAMEYCSGGDLRKLLNKPENCCGLK

[0479] ESQILSLLSDIGSGIRYLHENKIIHRDLKPENIVLQDVGGKIIHKIIDLGYAKDVDQGSLCTS FVGTLQYLAPELFENKPYTATVDYWSFGTMVFECIAGYRPFLHHLQPFTWHEKIKKKDP KCIF ACEEMSGE VRF S SHLPQPNSLC SLVVEPMENWLQLMLNWDPQQRGGP VDLTLKQ PRCFVLMDHILNLKIVHILNMTSAKIISFLLPPDESLHSLQSRIERETGINTGSQELLSETGIS LDPRKPASQCVLDGVRGCDSYMVYLFDKSKTVYEGPFASRSLSDCVNYIVQDSKIQLPII QLRKVWAEAVHYVSGLKEDYSRLFQGQRAAMLSLLRYNANLTKMKNTLISASQQLKA KLEFFHKSIQLDLERYSEQMTYGISSEKMLKAWKEMEEKAIHYAEVGVIGYLEDQIMSL HAEIMELQK SP YGRRQGDLMESLEQRAIDL YKQLKHRP SDHS YSD STEMVKIIVHT VQ S QDRVLKELFGHLSKLLGCKQKIIDLLPKVEVALSNIKEADNTVMFMQGKRQKEIWHLLK IACTQSSARSLVGSSLEGAVTPQTSAWLPPTSAEHDHSLSCVVTPQDGETSAQMIEENLN CLGHLSTIIHEANEEQGNSMMNLDWSWLTE SEQ ID NO: 158. IKKB MSWSPSLTTQTCGAWEMKERLGTGGFGNVIRWHNQETGEQIAIKQCRQELSPRNRERW CLEIQIMRRLTHPNVVAARDVPEGMQNLAPNDLPLLAMEYCQGGDLRKYLNQFENCCG LREGAILTLLSDIASALRYLHENRIIHRDLKPENIVLQQGEQRLIHKIIDLGYAKELDQGSL CTSFVGTLQYLAPELLEQQKYTVTVDYWSFGTLAFECITGFRPFLPNWQPVQWHSKVR QKSEVDIVVSEDLNGTVKF S S SLPYPNNLNS VLAERLEKWLQLMLMWHPRQRGTDPTY GPNGCFKALDDILNLKLVHILNMVTGTIHTYPVTEDESLQSLKARIQQDTGIPEEDQELL QEAGLALIPDKPATQCISDGKLNEGHTLDMDLVFLFDNSKITYETQISPRPQPESVSCILQ EPKRNLAFFQLRKVWGQVWHSIQTLKEDCNRLQQGQRAAMMNLLRNNSCLSKMKNS MASMSQQLKAKLDFFKT SIQIDLEKYSEQTEFGIT SDKLLL AWREMEQ AVELC GRENEV KLLVERMMALQTDIVDLQRSPMGRKQGGTLDDLEEQARELYRRLREKPRDQRTEGDSQ EMVRLLLQAIQSFEKKVRVIYTQLSKTVVCKQKALELLPKVEEVVSLMNEDEKTVVRLQ EKRQKELWNLLKIACSKVRGPVSGSPDSMNASRLSQPGQLMSQPSTASNSLPEPAKKSE ELVAEAHNLCTLLENAIQDTVREQDQSFTALDWSWLQTEEEEHSCLEQAS SEQ ID NO: 159. IKBA MFQAAERPQEWAMEGPRDGLKKERLLDDRHDSGLDSMKDEEYEQMVKELQEIRLEPQ EVPRGSEPWKQQLTEDGDSFLHLAIIHEEKALTMEVIRQVKGDLAFLNFQNNLQQTPLH LAVITNQPEIAEALLGAGCDPELRDFRGNTPLHLACEQGCLASVGVLTQSCTTPHLHSIL KATNYNGHTCLHLASIHGYLGIVELLVSLGADVNAQEPCNGRTALHLAVDLQNPDLVS LLLKCGADVNRVTYQGYSPYQLTWGRPSTRIQQQLGQLTLENLQMLPESEDEESYDTES EFTEFTEDELPYDDCVFGGQRLTL SEQ ID NO: 160. NF-kB

[0480] MAEDDPYLGRPEQMFHLDPSLTHTIFNPEVFQPQMALPTDGPYLQILEQPKQRGFRFRY VCEGPSHGGLPGASSEKNKKSYPQVKICNYVGPAKVIVQLVTNGKNIHLHAHSLVGKH CEDGICTVTAGPKDMVVGFANLGILHVTKKKVFETLEARMTEACIRGYNPGLLVHPDLA YLQAEGGGDRQLGDREKELIRQAALQQTKEMDLSVVRLMFTAFLPDSTGSFTRRLEPV VSDAIYDSKAPNASNLKIVRMDRTAGCVTGGEEIYLLCDKVQKDDIQIRFYEEEENGGV WEGFGDFSPTDVHRQFAIVFKTPKYKDINITKPASVFVQLRRKSDLETSEPKPFLYYPEIK DKEEVQRKRQKLMPNFSDSFGGGSGAGAGGGGMFGSGGGGGGTGSTGPGYSFPHYGF PTYGGITFHPGTTKSNAGMKHGTMDTESKKDPEGCDKSDDKNTVNLFGKVIETTEQDQ EPSEATVGNGEVTLTYATGTKEESAGVQDNLFLEKAMQLAKRHANALFDYAVTGDVK MLLAVQRHLTAVQDENGDSVLHLAIIHLHSQLVRDLLEVTSGLISDDIINMRNDLYQTPL HLAVITKQEDVVEDLLRAGADLSLLDRLGNSVLHLAAKEGHDKVLSILLKHKKAALLL DHPNGDGLNAIHLAMMSNSLPCLLLLVAAGADVNAQEQKSGRTALHLAVEHDNISLAG CLLLEGDAHVDSTTYDGTTPLHIAAGRGSTRLAALLKAAGADPLVENFEPLYDLDDSWE NAGEDEGVVPGTTPLDMAT S WQ VFDILNGKP YEPEFT SDDLL AQGDMKQLAED VKLQL YKLLEIPDPDKNWATLAQKLGLGILNNAFRLSPAPSKTLMDNYEVSGGTVRELVEALRQ MGYTEAIEVIQAASSPVKTTSQAHSLPLSPASTRQQIDELRDSDSVCDSGVETSFRKLSFT ESLTSGASLLTLNKMPHDYGQEGPLEGKI

[0481] SEQ ID NO: 16E PKC-delta

[0482] MAPFLRIAFNSYELGSLQAEDEANQPFCAVKMKEALSTERGKTLVQKKPTMYPEWKST FDAHIYEGRVIQIVLMRAAEEPVSEVTVGVSVLAERCKKNNGKAEFWLDLQPQAKVLM SVQYFLEDVDCKQSMRSEDEAKFPTMNRRGAIKQAKIHYIKNHEFIATFFGQPTFCSVCK DFVWGLNKQGYKCRQCNAAIHKKCIDKIIGRCTGTAANSRDTIFQKERFNIDMPHRFKV HNYMSPTFCDHCGSLLWGLVKQGLKCEDCGMNVHHKCREKVANLCGINQKLLAEALN QVTQRASRRSDSASSEPVGIYQGFEKKTGVAGEDMQDNSGTYGKIWEGSSKCNINNFIF HKVLGKGSFGKVLLGELKGRGEYFAIKALKKDVVLIDDDVECTMVEKRVLTLAAENPF LTHLICTFQTKDHLFFVMEFLNGGDLMYHIQDKGRFELYRATFYAAEIMCGLQFLHSKG IIYRDLKLDNVLLDRDGHIKIADFGMCKENIFGESRASTFCGTPDYIAPEILQGLKYTFSV DWWSFGVLLYEMLIGQSPFHGDDEDELFESIRVDTPHYPRWITKESKDILEKLFEREPTK RLGVTGNIKIHPFFKTINWTLLEKRRLEPPFRPKVKSPRDYSNFDQEFLNEKARLSYSDK NLIDSMDQS AF AGF SF VNPKFEHLLED

[0483] SEQ ID NO: 162. PI3K

[0484] MPPRPSSGELWGIHLMPPRILVECLLPNGMIVTLECLREATLITIKHELFKEARKYPLHQL LQDESSYIFVSVTQEAEREEFFDETRRLCDLRLFQPFLKVIEPVGNREEKILNREIGFAIGM PVCEFDMVKDPEVQDFRRNILNVCKEAVDLRDLNSPHSRAMYVYPPNVESSPELPKHIY NKLDKGQIIVVIWVIVSPNNDKQKYTLKINHDCVPEQVIAEAIRKKTRSMLLSSEQLKLC VLEYQGKYILKVCGCDEYFLEKYPLSQYKYIRSCIMLGRMPNLMLMAKESLYSQLPMD CFTMPSYSRRISTATPYMNGETSTKSLWVINSALRIKILCATYVNVNIRDIDKIYVRTGIY HGGEPLCDNVNTQRVPCSNPRWNEWLNYDIYIPDLPRAARLCLSICSVKGRKGAKEEHC PLAWGNINLFDYTDTLVSGKMALNLWPVPHGLEDLLNPIGVTGSNPNKETPCLELEFDW F S S WKFPDMS VIEEHANW S VSREAGF S YSHAGL SNRL ARDNELRENDKEQLKAISTRD PLSEITEQEKDFLWSHRHYCVTIPEILPKLLLSVKWNSRDEVAQMYCLVKDWPPIKPEQA MELLDCNYPDPMVRGFAVRCLEKYLTDDKLSQYLIQLVQVLKYEQYLDNLLVRFLLKK ALTNQRIGHFFFWHLKSEMHNKTVSQRFGLLLESYCRACGMYLKHLNRQVEAMEKLIN LTDILKQEKKDETQKVQMKFLVEQMRRPDFMDALQGFLSPLNPAHQLGNLRLEECRIM SSAKRPLWLNWENPDIMSELLFQNNEIIFKNGDDLRQDMLTLQIIRIMENIWQNQGLDLR MLPYGCLSIGDCVGLIEVVRNSHTIMQIQCKGGLKGALQFNSHTLHQWLKDKNKGEIYD AAIDLFTRSCAGYCVATFILGIGDRHNSNIMVKDDGQLFHIDFGHFLDHKKKKFGYKRE RVPFVLTQDFLIVISKGAQECTKTREFERFQEMCYKAYLAIRQHANLFINLFSMMLGSG MPELQSFDDIAYIRKTLALDKTEQEALEYFMKQMNDAHHGGWTTKMDWIFHTIKQHAL N

[0485] SEQ ID NO: 163. ERK1

[0486] MAAAAAQGGGGGEPRRTEGVGPGVPGEVEMVKGQPFDVGPRYTQLQYIGEGAYGMV SSAYDHVRKTRVAIKKISPFEHQTYCQRTLREIQILLRFRHENVIGIRDILRASTLEAMRD VYIVQDLMETDLYKLLKSQQLSNDHICYFLYQILRGLKYIHSANVLHRDLKPSNLLINTT CDLKICDFGLARIADPEHDHTGFLTEYVATRWYRAPEIMLNSKGYTKSIDIWSVGCILAE MLSNRPIFPGKHYLDQLNHILGILGSPSQEDLNCIINMKARNYLQSLPSKTKVAWAKLFP KSDSKALDLLDRMLTFNPNKRITVEEALAHPYLEQYYDPTDEPVAEEPFTFAMELDDLP KERLKELIFQETARFQPGVLEAP SEQ ID NO: 164. ERK2

[0487] MAAAAAAGAGPEMVRGQVFDVGPRYTNLSYIGEGAYGMVCSAYDNVNKVRVAIKKIS PFEHQTYCQRTLREIKILLRFRHENIIGINDIIRAPTIEQMKDVYIVQDLMETDLYKLLKTQ HLSNDHICYFLYQILRGLKYIHSANVLHRDLKPSNLLLNTTCDLKICDFGLARVADPDHD HTGFLTEYVATRWYRAPEIMLNSKGYTKSIDIWSVGCILAEMLSNRPIFPGKHYLDQLNH ILGILGSPSQEDLNCIINLKARNYLLSLPHKNKVPWNRLFPNADSKALDLLDKMLTFNPH KRIEVEQALAHPYLEQYYDPSDEPIAEAPFKFDMELDDLPKEKLKELIFEETARFQPGYR S

[0488] SEQ ID NO: 165. Raf-1 MEHIQGAWKTISNGFGFKDAVFDGSSCISPTIVQQFGYQRRASDDGKLTDPSKTSNTIRV

[0489] FLPNKQRTVVNVRNGMSLHDCLMKALKVRGLQPECCAVFRLLHEHKGKKARLDWNT

[0490] DAASLIGEELQVDFLDHVPLTTHNFARKTFLKLAFCDICQKFLLNGFRCQTCGYKFHEH

[0491] CSTKVPTMCVDWSNIRQLLLFPNSTIGDSGVPALPSLTMRRMRESVSRMPVSSQHRYSTP

[0492] HAFTFNTSSPSSEGSLSQRQRSTSTPNVHMVSTTLPVDSRMIEDAIRSHSESASPSALSSSP

[0493] NNLSPTGWSQPKTPVPAQRERAPVSGTQEKNKIRPRGQRDSSYYWEIEASEVMLSTRIGS

[0494] GSFGTVYKGKWHGDVAVKILKVVDPTPEQFQAFRNEVAVLRKTRHVNILLFMGYMTK

[0495] DNLAIVTQWCEGSSLYKHLHVQETKFQMFQLIDIARQTAQGMDYLHAKNIIHRDMKSN

[0496] NIFLHEGLTVKIGDFGLATVKSRWSGSQQVEQPTGSVLWMAPEVIRMQDNNPFSFQSDV

[0497] YSYGIVLYELMTGELPYSHINNRDQIIFMVGRGYASPDLSKLYKNCPKAMKRLVADCVK

[0498] KVKEERPLFPQILSSIELLQHSLPKINRSASEPSLHRAAHTEDINACTLTTSPRLPVF

[0499] SEQ ID NO: 166. Pakl

[0500] MSNNGLDIQDKPPAPPMRNTSTMIGAGSKDAGTLNHGSKPLPPNPEEKKKKDRFYRSIL

[0501] PGDKTNKKKEKERPEISLPSDFEHTIHVGFDAVTGEFTGMPEQWARLLQTSNITKSEQKK

[0502] NPQAVLDVLEFYNSKKTSNSQKYMSFTDKSAEDYNSSNALNVKAVSETPAVPPVSEDE

[0503] DDDDDDATPPPVIAPRPEHTKSVYTRSVIEPLPVTPTRDVATSPISPTENNTTPPDALTRNT

[0504] EKQKKKPKMSDEEILEKLRSIVSVGDPKKKYTRFEKIGQGASGTVYTAMDVATGQEVAI

[0505] KQMNLQQQPKKELIINEILVMRENKNPNIVNYLDSYLVGDELWVVMEYLAGGSLTDVV

[0506] TETCMDEGQIAAVCRECLQALEFLHSNQVIHRDIKSDNILLGMDGSVKLTDFGFCAQITP

[0507] EQSKRSTMVGTPYWMAPEVVTRKAYGPKVDIWSLGIMAIEMIEGEPPYLNENPLRALYL

[0508] IATNGTPELQNPEKLSAIFRDFLNRCLEMDVEKRGSAKELLQHQFLKIAKPLSSLTPLIAA

[0509] AKEATKNNH

[0510] SEQ ID NO: 167. Rael

[0511] MQAIKCVVVGDGAVGKTCLLISYTTNAFPGEYIPTVFDNYSANVMVDGKPVNLGLWDT

[0512] AGQEDYDRLRPLSYPQTDVFLICFSLVSPASFENVRAKWYPEVRHHCPNTPIILVGTKLD

[0513] LRDDKDTIEKLKEKKLTPITYPQGLAMAKEIGAVKYLECSALTQRGLKTVFDEAIRAVLC

[0514] PPPVKKRKRKCLLL

[0515] SEQ ID NO: 168. Rac2

[0516] MQAIKCVVVGDGAVGKTCLLISYTTNAFPGEYIPTVFDNYSANVMVDSKPVNLGLWDT

[0517] AGQEDYDRLRPLSYPQTDVFLICFSLVSPASYENVRAKWFPEVRHHCPSTPIILVGTKLD LRDDKDTIEKLKEKKLAPITYPQGLALAKEIDSVKYLECSALTQRGLKTVFDEAIRAVLC

[0518] PQPTRQQKRACSLL

[0519] SEQ ID NO: 169. Rac3

[0520] MQAIKCVVVGDGAVGKTCLLISYTTNAFPGEYIPTVFDNYSANVMVDGKPVNLGLWDT

[0521] AGQEDYDRLRPLSYPQTDVFLICFSLVSPASFENVRAKWYPEVRHHCPHTPILLVGTKLD

[0522] LRDDKDTIERLRDKKLAPITYPQGLAMAREIGSVKYLECSALTQRGLKTVFDEAIRAVLC

[0523] PPPVKKPGKKCTVF

[0524] SEQ ID NO: 170. RhoA

[0525] MAAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENYVADIEVDGKQVELALW

[0526] DTAGQEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVKHFCPNVPIILVGNKK

[0527] DLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKTKDGVREVFEMATR

[0528] AALQARRGKKKSGCLVL

[0529] SEQ ID NO: 171. ROCK1

[0530] MSTGDSFETRFEKMDNLLRDPKSEVNSDCLLDGLDALVYDLDFPALRKNKNIDNFLSRY

[0531] KDTINKIRDLRMKAEDYEVVKVIGRGAFGEVQLVRHKSTRKVYAMKLLSKFEMIKRSD

[0532] SAFFWEERDIMAFANSPWVVQLFYAFQDDRYLYMVMEYMPGGDLVNLMSNYDVPEK

[0533] WARFYTAEVVLALDAIHSMGFIHRDVKPDNMLLDKSGHLKLADFGTCMKMNKEGMV

[0534] RCDTAVGTPDYISPEVLKSQGGDGYYGRECDWWSVGVFLYEMLVGDTPFYADSLVGT

[0535] YSKIMNHKNSLTFPDDNDISKEAKNLICAFLTDREVRLGRNGVEEIKRHLFFKNDQWAW

[0536] ETLRDTVAPVVPDLSSDIDTSNFDDLEEDKGEEETFPIPKAFVGNQLPFVGFTYYSNRRY

[0537] LSSANPNDNRTSSNADKSLQESLQKTIYKLEEQLHNEMQLKDEMEQKCRTSNIKLDKIM

[0538] KELDEEGNQRRNLESTVSQIEKEKMLLQHRINEYQRKAEQENEKRRNVENEVSTLKDQL

[0539] EDLKKVSQNSQLANEKLSQLQKQLEEANDLLRTESDTAVRLRKSHTEMSKSISQLESLN

[0540] RELQERNRILENSKSQTDKDYYQLQAILEAERRDRGHDSEMIGDLQARITSLQEEVKHLK

[0541] HNLEKVEGERKEAQDMLNHSEKEKNNLEIDLNYKLKSLQQRLEQEVNEHKVTKARLTD

[0542] KHQSIEEAKSVAMCEMEKKLKEEREAREKAENRVVQIEKQCSMLDVDLKQSQQKLEHL

[0543] TGNKERMEDEVKNLTLQLEQESNKRLLLQNELKTQAFEADNLKGLEKQMKQEINTLLE

[0544] AKRLLEFELAQLTKQYRGNEGQMRELQDQLEAEQYFSTLYKTQVKELKEEIEEKNRENL

[0545] KKIQELQNEKETL ATQLDL AETKAE SEQL ARGLLEEQ YFELTQE SKKAASRNRQEITDK

[0546] DHTVSRLEEANSMLTKDIEILRRENEELTEKMKKAEEEYKLEKEEEISNLKAAFEKNINT

[0547] ERTLKTQAVNKLAEIMNRKDFKIDRKKANTQDLRKKEKENRKLQLELNQEREKFNQMV VKHQKELNDMQAQLVEECAITRNELQMQLASKESDIEQLRAKLLDLSDSTSVASFPSAD ETDGNLPE SRIEGWLS VPNRGNIKRYGWKKQ YVVVS SKKILF YNDEQDKEQ SNP SMVL DIDKLFHVRPVTQGDVYRAETEEIPKIFQILYANEGECRKDVEMEPVQQAEKTNFQNHK GHEFIPTLYHFPANCDACAKPLWHVFKPPPALECRRCHVKCHRDHLDKKEDLICPCKVS YDVTSARDMLLLACSQDEQKKWVTHLVKKIPKNPPSGFVRASPRTLSTRSTANQSFRKV VKNTSGKTS

[0548] SEQ ID NO: 172. ROCK2

[0549] MSRPPPTGKMPGAPETAPGDGAGASRQRKLEALIRDPRSPINVESLLDGLNSLVLDLDFP ALRKNKNIDNFLNRYEKIVKKIRGLQMKAEDYDVVKVIGRGAFGEVQLVRHKASQKVY AMKLLSKFEMIKRSDSAFFWEERDIMAFANSPWVVQLFYAFQDDRYLYMVMEYMPGG DLVNLMSNYDVPEI<WAI<FYTAEVVLALDAIHSMGL1HRDVKPDNMLLD1<HGHLI<LAD FGTCMKMDETGMVHCDTAVGTPDYISPEVLKSQGGDGFYGRECDWWSVGVFLYEML VGDTPFYADSLVGTYSKIMDHKNSLCFPEDAEISKHAKNLICAFLTDREVRLGRNGVEEI RQHPFFKNDQWHWDNIRET AAP VVPEL S SDID S SNFDDIEDDKGD VETFPIPKAF VGNQL PFIGFT YYRENLLL SD SP SCRETD SIQ SRKNEE SQEIQKKLYTLEEHL SNEMQ AKEELEQK CKSVNTRLEKTAKELEEEITLRKSVESALRQLEREKALLQHKNAEYQRKADHEADKKR NLENDVNSLKDQLEDLKKRNQNSQISTEKVNQLQRQLDETNALLRTESDTAARLRKTQ AES SKQIQQLE SNNRDLQDKNCLLETAKLKLEKEFINLQ SALE SERRDRTHGSEIINDLQG RICGLEEDLKNGKILLAKVELEKRQLQERFTDLEKEKSNMEIDMTYQLKVIQQSLEQEEA EHKATKARLADKNKIYESIEEAKSEAMKEMEKKLLEERTLKQKVENLLLEAEKRCSLLD CDLKQSQQKINELLKQKDVLNEDVRNLTLKIEQETQKRCLTQNDLKMQTQQVNTLKMS EKQLKQENNHLMEMKMNLEKQNAELRKERQDADGQMKELQDQLEAEQYFSTLYKTQ VRELKEECEEKTKLGKELQQKKQELQDERDSLAAQLEITLTKADSEQLARSIAEEQYSDL EKEKIMKELEIKEMMARHKQELTEKDATIASLEETNRTLTSDVANLANEKEELNNKLKD VQEQLSRLKDEEISAAAIKAQFEKQLLTERTLKTQAVNKLAEIMNRKEPVKRGNDTDVR RKEKENRKLHMELKSEREKLTQQMIKYQKELNEMQAQIAEESQIRIELQMTLDSKDSDI EQLRSQLQALHIGLDS S SIGSGPGDAEADDGFPESRLEGWLSLPVRNNTKKFGWVKKYV IVSSKKILFYDSEQDKEQSNPYMVLDIDKLFHVRPVTQTDVYRADAKEIPRIFQILYANEG ESKKEQEFPVEPVGEKSNYICHKGHEFIPTLYHFPTNCEACMKPLWHMFKPPPALECRRC HIKCHKDHMDKKEEIIAPCKVYYDISTAKNLLLLANSTEEQQKWVSRLVKKIPKKPPAP DPF ARS SPRT SMKIQQNQ SIRRP SRQL APNKP S SEQ ID NO: 173. Alpha-tubulin

[0550] MRECISIHVGQ AGVQIGNAC WEL YCLEHGIQPDGQMP SDKTIGGGDD SFNTFF SETGAG KHVPRAVFVDLEPTVIDEVRTGTYRQLFHPEQLITGKEDAANNYARGHYTIGKEIIDLVL DRIRKLADQCTGLQGFLVFHSFGGGTGSGFTSLLMERLSVDYGKKSKLEFSIYPAPQVST AVVEPYNSILTTHTTLEHSDCAFMVDNEAIYDICRRNLDIERPTYTNLNRLIGQIVSSITAS LRFDGALNVDLTEFQTNLVPYPRIHFPLATYAPVISAEKAYHEQLSVAEITNACFEPANQ MVKCDPRHGKYMACCLLYRGDVVPKDVNAAIATIKTKRTIQFVDWCPTGFKVGINYQP PTVVPGGDLAKVQRAVCMLSNTTAIAEAWARLDHKFDLMYAKRAFVHWYVGEGMEE GEFSEAREDMAALEKDYEEVGVDSVEGEGEEEGEEY

[0551] SEQ ID NO: 174. Beta-tubulin

[0552] MREIVHIQIGQCGNQIGAKFWEMIGEEHGIDLAGSDRGASALQLERISVYYNEAYGRKY VPRAVLVDLEPGTMDSIRSSKLGALFQPDSFVHGNSGAGNNWAKGHYTEGAELIENVL EVVRHESESCDCLQGFQIVHSLGGGTGSGMGTLLMNKIREEYPDRIMNSFSVMPSPKVS DTVVEPYNAVLSIHQLIENADACFCIDNEALYDICFRTLKLTTPTYGDLNHLVSLTMSGIT TSLRFPGQLNADLRKLAVNMVPFPRLHFFMPGFAPLTAQGSQQYRALSVAELTQQMFD ARNTMAACDLRRGRYLTVACIFRGKMSTKEVDQQLLSVQTRNSSCFVEWIPNNVKVAV CDIPPRGLSMAATFIGNNTAIQEIFNRVSEHFSAMFKRKAFVHWYTSEGMDINEFGEAEN NIHDLVSEYQQFQDAKAVLEEDEEVTEEAEMEPEDKGH

[0553] SEQ ID NO: 175. Vimentin

[0554] MSTRSVSSSSYRRMFGGPGTASRPSSSRSYVTTSTRTYSLGSALRPSTSRSLYASSPGGVY ATRS S AVRLRS S VPGVRLLQD S VDF SLAD AINTEFKNTRTNEKVELQELNDRF ANYIDK VRFLEQQNKILLAELEQLKGQGKSRLGDLYEEEMRELRRQVDQLTNDKARVEVERDNL AEDIMRLREKLQEEMLQREEAENTLQSFRQDVDNASLARLDLERKVESLQEEIAFLKKL HEEEIQELQAQIQEQHVQIDVDVSKPDLTAALRDVRQQYESVAAKNLQEAEEWYKSKF ADLSEAANRNNDALRQAKQESTEYRRQVQSLTCEVDALKGTNESLERQMREMEENFA VEAANYQDTIGRLQDEIQNMKEEMARHLREYQDLLNVKMALDIEIATYRKLLEGEESRI SLPLPNFSSLNLRETNLDSLPLVDTHSKRTLLIKTVETRDGQVINETSQHHDDLE Ion channels

[0555] SEQ ID NO: 178. ATP1A1

[0556] MGKGVGRDKYEPAAVSEQGDKKGKKGKKDRDMDELKKEVSMDDHKLSLDELHRKY GTDLSRGLTSARAAEILARDGPNALTPPPTTPEWIKFCRQLFGGFSMLLWIGAILCFLAYS IQAATEEEPQNDNLYLGVVLSAVVIITGCFSYYQEAKSSKIMESFKNMVPQQALVIRNGE KMSINAEEVVVGDLVEVKGGDRIPADLRIISANGCKVDNSSLTGESEPQTRSPDFTNENP LETRNIAFFSTNCVEGTARGIVVYTGDRTVMGRIATLASGLEGGQTPIAAEIEHFIHIITGV AVFLGVSFFILSLILEYTWLEAVIFLIGIIVANVPEGLLATVTVCLTLTAKRMARKNCLVK NLEAVETLGST STIC SDKTGTLTQNRMT VAHMWFDNQIHEADTTENQ SGVSFDKT S AT WLALSRIAGLCNRAVFQANQENLPILKRAVAGDASESALLKCIELCCGSVKEMRERYAK IVEIPFNSTNKYQLSIHKNPNTSEPQHLLVMKGAPERILDRCSSILLHGKEQPLDEELKDA FQNAYLELGGLGERVLGFCHLFLPDEQFPEGFQFDTDDVNFPIDNLCFVGLISMIDPPRA AVPDAVGKCRSAGIKVIMVTGDHPITAKAIAKGVGIISEGNETVEDIAARLNIPVSQVNPR D AKAC VVHGSDLKDMT SEQLDDILKYHTEIVF ART SPQQKLIIVEGCQRQGAIVAVTGD GVNDSPALKKADIGVAMGIAGSDVSKQAADMILLDDNFASIVTGVEEGRLIFDNLKKSI AYTLTSNIPEITPFLIFIIANIPLPLGTVTILCIDLGTDMVPAISLAYEQAESDIMKRQPRNPK TDKLVNERLISMAYGQIGMIQALGGFFTYFVILAENGFLPIHLLGLRVDWDDRWINDVE DSYGQQWTYEQRKIVEFTCHTAFFVSIVVVQWADLVICKTRRNSVFQQGMKNKILIFGL

[0557] FEETALAAFLSYCPGMGVALRMYPLKPTWWFCAFPYSLLIFVYDEVRKLIIRRRPGGWV EKETYY

[0558] SEQ ID NO: 179. ATP1A2

[0559] MGRGAGREYSPAATTAENGGGKKKQKEKELDELKKEVAMDDHKLSLDELGRKYQVD LSKGLTNQRAQDVLARDGPNALTPPPTTPEWVKFCRQLFGGFSILLWIGAILCFLAYGIQ AAMEDEPSNDNLYLGVVLAAVVIVTGCFSYYQEAKSSKIMDSFKNMVPQQALVIREGE KMQINAEEVVVGDLVEVKGGDRVPADLRIIS SHGCKVDNS SLTGESEPQTRSPEFTHENP LETRNICFFSTNCVEGTARGIVIATGDRTVMGRIATLASGLEVGRTPIAMEIEHFIQLITGV AVFLGVSFFVLSLILGYSWLEAVIFLIGIIVANVPEGLLATVTVCLTLTAKRMARKNCLV KNLEAVETLGSTSTICSDKTGTLTQNRMTVAHMWFDNQIHEADTTEDQSGATFDKRSPT WTALSRIAGLCNRAVFKAGQENISVSKRDTAGDASESALLKCIELSCGSVRKMRDRNPK VAEIPFNSTNKYQLSIHEREDSPQSHVLVMKGAPERILDRCSTILVQGKEIPLDKEMQDAF QNAYMELGGLGERVLGFCQLNLPSGKFPRGFKFDTDELNFPTEKLCFVGLMSMIDPPRA AVPDAVGKCRSAGIKVIMVTGDHPITAKAIAKGVGIISEGNETVEDIAARLNIPMSQVNP REAKACVVHGSDLKDMTSEQLDEILKNHTEIVFARTSPQQKLIIVEGCQRQGAIVAVTGD GVNDSPALKKADIGIAMGISGSDVSKQAADMILLDDNFASIVTGVEEGRLIFDNLKKSIA YTLTSNIPEITPFLLFIIANIPLPLGTVTILCIDLGTDMVPAISLAYEAAESDIMKRQPRNSQT DKLVNERLISMAYGQIGMIQALGGFFTYFVILAENGFLPSRLLGIRLDWDDRTMNDLEDS YGQEWTYEQRKVVEFTCHTAFFASIVVVQWADLIICKTRRNSVFQQGMKNKILIFGLLE ETALAAFLSYCPGMGVALRMYPLKVTWWFCAFPYSLLIFIYDEVRKLILRRYPGGWVE KETYY

[0560] SEQ ID NO: 180. ATP1A3

[0561] MGDKKDDKDSPKKNKGKERRDLDDLKKEVAMTEHKMSVEEVCRKYNTDCVQGLTHS KAQEILARDGPNALTPPPTTPEWVKFCRQLFGGFSILLWIGAILCFLAYGIQAGTEDDPSG DNLYLGIVLAAVVIITGCFSYYQEAKSSKIMESFKNMVPQQALVIREGEKMQVNAEEVV VGDLVEIKGGDRVPADLRIIS AHGCKVDNS SLTGESEPQTRSPDCTHDNPLETRNITFF ST NCVEGTARGVVVATGDRTVMGRIATLASGLEVGKTPIAIEIEHFIQLITGVAVFLGVSFFI LSLILGYTWLEAVIFLIGIIVANVPEGLLATVTVCLTLTAKRMARKNCLVKNLEAVETLG STSTICSDKTGTLTQNRMTVAHMWFDNQIHEADTTEDQSGTSFDKSSHTWVALSHIAGL CNRAVFKGGQDNIPVLKRDVAGDASESALLKCIELSSGSVKLMRERNKKVAEIPFNSTN KYQLSIHETEDPNDNRYLLVMKGAPERILDRCSTILLQGKEQPLDEEMKEAFQNAYLEL GGLGERVLGFCHYYLPEEQFPKGFAFDCDDVNFTTDNLCFVGLMSMIDPPRAAVPDAV GKCRSAGIKVIMVTGDHPITAKAIAKGVGIISEGNETVEDIAARLNIPVSQVNPRDAKAC VIHGTDLKDFTSEQIDEILQNHTEIVFARTSPQQKLIIVEGCQRQGAIVAVTGDGVNDSPA LKKADIGVAMGIAGSDVSKQAADMILLDDNFASIVTGVEEGRLIFDNLKKSIAYTLTSNI PEITPFLLFIMANIPLPLGTITILCIDLGTDMVPAISLAYEAAESDIMKRQPRNPRTDKLVNE RLISMAYGQIGMIQALGGFFSYFVILAENGFLPGNLVGIRLNWDDRTVNDLEDSYGQQW TYEQRKVVEFTCHTAFFVSIVVVQWADLIICKTRRNSVFQQGMKNKILIFGLFEETALAA FLSYCPGMDVALRMYPLKPSWWFCAFPYSFLIFVYDEIRKLILRRNPGGWVEKETYY SEQ ID NO: 181. ATP1A4

[0562] MGLWGKKGTVAPHDQSPRRRPKKGLIKKKMVKREKQKRNMEELKKEVVMDDHKLTL EELSTKYSVDLTKGHSHQRAKEILTRGGPNTVTPPPTTPEWVKFCKQLFGGFSLLLWTG AILCFVAYSIQIYFNEEPTKDNLYLSIVLSVVVIVTGCFSYYQEAKSSKIMESFKNMVPQQ ALVIRGGEKMQINVQEVVLGDLVEIKGGDRVP ADLRLIS AQGCKVDNS SLTGESEPQ SR SPDFTHENPLETRNICFFSTNCVEGTARGIVIATGDSTVMGRIASLTSGLAVGQTPIAAEIE HFIHLITVVAVFLGVTFFALSLLLGYGWLEAUFLIGIIVANVPEGLLATVTVCLTLTAKRM ARKNCLVKNLEAVETLGSTSTICSDKTGTLTQNRMTVAHMWFDMTVYEADTTEEQTG KTFTKSSDTWFMLARIAGLCNRADFKANQEILPIAKRATTGDASESALLKFIEQSYSSVA EMREKNPKVAEIPFNSTNKYQMSIHLREDSSQTHVLMMKGAPERILEFCSTFLLNGQEYS MNDEMKE AFQNAYLELGGLGERVLGFCFLNLP S SF SKGFPFNTDEINFPMDNLCF VGLIS MIDPPRAAVPDAVSKCRSAGIKVIMVTGDHPITAKAIAKGVGIISEGTETAEEVAARLKIP ISKVDASAAKAIVVHGAELKDIQSKQLDQILQNHPEIVFARTSPQQKLIIVEGCQRLGAVV AVTGDGVNDSPALKKADIGIAMGISGSDVSKQAADMILLDDNFASIVTGVEEGRLIFDNL KKSIMYTLTSNIPEITPFLMFIILGIPLPLGTITILCIDLGTDMVPAISLAYESAESDIMKRLP RNPKTDNLVNHRLIGMAYGQIGMIQALAGFFTYFVILAENGFRPVDLLGIRLHWEDKYL NDLEDSYGQQWTYEQRKVVEFTCQTAFFVTIVVVQWADLIISKTRRNSLFQQGMRNKV LIFGILEETLLAAFLSYTPGMDVALRMYPLKITWWLCAIPYSILIFVYDEIRKLLIRQHPDG WVERETYY

[0563] SEQ ID NO: 182. ATP1B1

[0564] MARGKAKEEGSWKKFIWNSEKKEFLGRTGGSWFKILLFYVIFYGCLAGIFIGTIQVMLLT ISEFKPTYQDRVAPPGLTQIPQIQKTEISFRPNDPKSYEAYVLNIVRFLEKYKDSAQRDDM IFEDCGDVPSEPKERGDFNHERGERKVCRFKLEWLGNCSGLNDETYGYKEGKPCIIIKLN RVLGFKPKPPKNESLETYPVMKYNPNVLPVQCTGKRDEDKDKVGNVEYFGLGNSPGFP LQYYPYYGKLLQPKYLQPLLAVQFTNLTMDTEIRIECKAYGENIGYSEKDRFQGRFDVKI EVKS

[0565] SEQ ID NO: 183. ATP1B2

[0566] MVIQKEKKSCGQVVEEWKEFVWNPRTHQFMGRTGTSWAFILLFYLVFYGFLTAMFTLT MWVMLQTVSDHTPKYQDRLATPGLMIRPKTENLDVIVNVSDTESWDQHVQKLNKFLE PYNDSIQAQKNDVCRPGRYYEQPDNGVLNYPKRACQFNRTQLGNCSGIGDSTHYGYST GQPCVFIKMNRVINFYAGANQSMNVTCAGKRDEDAENLGNFVMFPANGNIDLMYFPY YGKKFHVNYTQPLVAVKFLNVTPNVEVNVECRINAANIATDDERDKFAGRVAFKLRIN KT

[0567] SEQ ID NO: 184. ATP1B3

[0568] MTKNEKKSLNQSLAEWKLFIYNPTTGEFLGRTAKSWGLILLFYLVFYGFLAALFSFTMW VMLQTLNDEVPKYRDQIP SPGLMVFPKP VT ALEYTF SRSDPT S YAGYIEDLKKFLKP YTL EEQKNLTVCPDGALFEQKGPVYVACQFPISLLQACSGMNDPDFGYSQGNPCILVKMNRI IGLKPEGVPRIDCVSKNEDIPNVAVYPHNGMIDLKYFPYYGKKLHVGYLQPLVAVQVSF APNNTGKEVTVECKIDGSANLKSQDDRDKFLGRVMFKITARA

[0569] SEQ ID NO: 185. ATP1B4 MRRQLRSRRAPSFPYSYRYRLDDPDEANQNYLADEEEEAEEEARVTVVPKSEEEEEEEE KEEEEEEEKEEEEGQGQPTGNAWWQKLQIMSEYLWDPERRMFLARTGQSWSLILLIYFF FYASLAAVITLCMYTLFLTISPYIPTFTERVKPPGVMIRPFAHSLNFNFNVSEPDTWQHYV ISLNGFLQGYNDSLQEEMNVDCPPGQYFIQDGNEDEDKKACQFKRSFLKNCSGLEDPTF GYSTGQPCILLKMNRIVGFRPELGDPVKVSCKVQRGDENDIRSISYYPESASFDLRYYPY YGKLTHVNYTSPLVAMHFTDVVKNQAVPVQCQLKGKGVINDVINDRFVGRVIFTLNIET SEQ ID NO: 186. SLC12A1

[0570] MSLNNSSNVFLDSVPSNTNRFQVSVINENHESSAAADDNTDPPHYEETSFGDEAQKRLRI SFRPGNQECYDNFLQSGETAKTDASFHAYDSHTNTYYLQTFGHNTMDAVPKIEYYRNT GSISGPKVNRPSLLEIHEQLAKNVAVTPSSADRVANGDGIPGDEQAENKEDDQAGVVKF GWVKGVLVRCMLNIWGVMLFIRLSWIVGEAGIGLGVLIILLSTMVTSITGLSTSAIATNG FVRGGGAYYLISRSLGPEFGGSIGLIFAFANAVAVAMYVVGFAETVVDLLKESDSMMV DPTNDIRIIGSITVVILLGISVAGMEWEAKAQVILLVILLIAIANFFIGTVIPSNNEKKSRGFF NYQASIFAENFGPRFTKGEGFFSVFAIFFPAATGILAGANISGDLEDPQDAIPRGTMLAIFI

[0571] TTVAYLGVAICVGACVVRDATGNMNDTIISGMNCNGSAACGLGYDFSRCRHEPCQYGL MNNFQVMSMVSGFGPLITAGIF S ATLS S ALASLVS APKVFQALCKDNIYKALQFFAKGY GKNNEPLRGYILTFLIAMAFILIAELNTIAPIISNFFLASYALINFSCFHASYAKSPGWRPAY GIYNMWVSLFGAVLCCAVMFVINWWAAVITYVIEFFLYVYVTCKKPDVNWGSSTQALS YVSALDNALELTTVEDHVKNFRPQCIVLTGGPMTRPALLDITHAFTKNSGLCICCEVFVG PRKLCVKEMNSGMAKKQAWLIKNKIKAFYAAVAADCFRDGVRSLLQASGLGRMKPNT LVIGYKKNWRKAPLTEIENYVGIIHDAFDFEIGVVIVRISQGFDISQVLQVQEELERLEQE

[0572] RLALEATIKDNECEEESGGIRGLFKKAGKLNITKTTPKKDGSINTSQSMHVGEFNQKLVE ASTQFKKKQEKGTIDVWWLFDDGGLTLLIPYILTLRKKWKDCKLRIYVGGKINRIEEEKI VMASLLSKFRIKFADIHIIGD1NIRPNKESWKVFEEMIEPYRLHESCKDLTTAEKLKRETP WKITDAELEAVKEKSYRQVRLNELLQEHSRAANLIVLSLPVARKGSISDLLYMAWLEIL TKNLPPVLLVRGNHKNVLTFYS

[0573] SEQ ID NO: 187. SLC12A2

[0574] MEPRPTAPSSGAPGLAGVGETPSAAALAAARVELPGTAVPSVPEDAAPASRDGGGVRD EGPAAAGDGLGRPLGPTPSQSRFQVDLVSENAGRAAAAAAAAAAAAAAAGAGAGAK QTPADGEASGESEPAKGSEEAKGRFRVNFVDPAASSSAEDSLSDAAGVGVDGPNVSFQN GGDTVLSEGSSLHSGGGGGSGHHQHYYYDTHTNTYYLRTFGHNTMDAVPRIDHYRHT AAQLGEKLLRPSLAELHDELEKEPFEDGFANGEESTPTRDAVVTYTAESKGVVKFGWIK GVLVRCMLNIWGVMLFIRLSWIVGQAGIGLSVLVIMMATVVTTITGLSTSAIATNGFVR GGGAYYLISRSLGPEFGGAIGLIFAFANAVAVAMYVVGFAETVVELLKEHSILMIDEINDI RIIGAITVVILLGISVAGMEWEAKAQIVLLVILLLAIGDFVIGTFIPLESKKPKGFFGYKSEI FNENFGPDFREEETFFSVFAIFFPAATGILAGANISGDLADPQSAIPKGTLLAILITTLVYVG IAVSVGSCVVRDATGNVNDTIVTELTNCTSAACKLNFDFSSCESSPCSYGLMNNFQVMS MVSGFTPLIS AGIF S ATLS S AL ASLVS APKIFQALCKDNIYPAFQMF AKGYGKNNEPLRG YILTFLIALGFILIAELNVIAPIISNFFLASYALINFSVFHASLAKSPGWRPAFKYYNMWISL LGAILCCTVMFVINWWAALLTYVIVLGLYIYVTYKKPDVNWGSSTQALTYLNALQHSIR LSGVEDHVKNFRPQCLVMTGAPNSRP ALLHL VHDFTKNVGLMICGHVHMGPRRQAMK EMS1DQAKYQRWL1KNKMKAFYAPVHADDLREGAQYLMQAAGLGRMKPNTLVLGFK KDWLQ ADMRD VDM YINLFHD AFDIQ YGVVVIRLKEGLDISHLQGQEELL S SQEK SPGTK DVVVSVEYSKKSDLDTSKPLSEKPITHKVEEEDGKTATQPLLKKESKGPIVPLNVADQKL LEASTQFQKKQGKNTIDVWWLFDDGGLTLLIPYLLTTKKKWKDCKIRVFIGGKINRIDH DRRAMATLLSKFRIDFSDIMVLGDINTKPKKENIIAFEEIIEPYRLHEDDKEQDIADKMKE DEPWRITDNELELYKTKTYRQIRLNELLKEHSSTANIIVMSLPVARKGAVSSALYMAWL EALSKDLPPILLVRGNHQSVLTFYS SEQ ID NO: 188. SLC9A1

[0575] MVLRSGICGLSPHRIFPSLLVVVALVGLLPVLRSHGLQLSPTASTIRSSEPPRERSIGDVTT APPEVTPESRPVNHSVTDHGMKPRKAFPVLGIDYTHVRTPFEISLWILLACLMKIGFHVIP TISSIVPESCLLIVVGLLVGGLIKGVGETPPFLQSDVFFLFLLPPIILDAGYFLPLRQFTENL GTILIFAVVGTLWNAFFLGGLMYAVCLVGGEQINNIGLLDNLLFGSIISAVDPVAVLAVF EEIHINELLHILVFGESLLNDAVTVVLYHLFEEFANYEHVGIVDIFLGFLSFFVVALGGVL VGVVYGVIAAFTSRFTSHIRVIEPLFVFLYSYMAYLSAELFHLSGIMALIASGVVMRPYV EANISHKSHTTIKYFLKMWSSVSETLIFIFLGVSTVAGSHHWNWTFVISTLLFCLIARVLG VLGLTWFINKFRIVKLTPKDQFIIAYGGLRGAIAFSLGYLLDKKHFPMCDLFLTAIITVIFF TVFVQGMTIRPLVDLLAVKKKQETKRSINEEIHTQFLDHLLTGIEDICGHYGHHHWKDK LNRFNKKYVKKCLIAGERSKEPQLIAFYHKMEMKQAIELVESGGMGKIPSAVSTVSMQN IHPKSLP SERILP AL SKDKEEEIRKILRNNLQKTRQRLRS YNRHTLVADP YEEAWNQMLL RRQKARQLEQKINNYLT VP AHKLD SPTMSRARIGSDPL AYEPKEDLP VITIDP ASPQ SPES VDLVNEELKGKVLGLSRDPAKVAEEDEDDDGGIMMRSKETSSPGTDDVFTPAPSDSPSS QRIQRCL SDPGPHPEPGEGEPFFPKGQ SEQ ID NO: 189. SLC9A2

[0576] MEPLGNWRSLRAPLPPMLLLLLLQVAGPVGALAETLLNAPRAMGTSSSPPSPASVVAPG TTLFEESRLPVFTLDYPHVQIPFEITLWILLASLAKIGFHLYHKLPTIVPESCLLIMVGLLLG GIIFGVDEKSPPAMKTDVFFLYLLPPIVLDAGYFMPTRPFFENIGTIFWYAVVGTLWNSIG IGVSLFGICQIEAFGLSDITLLQNLLFGSLISAVDPVAVLAVFENIHVNEQLYILVFGESLLN DAVTVVLYNLFKSFCQMKTIETIDVFAGIANFFVVGIGGVLIGIFLGFIAAFTTRFTHNIRV IEPLFVFLYSYLSYITAEMFHLSGIMAITACAMTMNKYVEENVSQKSYTTIKYFMKMLSS VSETLIFIFMGVSTVGKNHEWNWAFVCFTLAFCLMWRALGVFVLTQVINRFRTIPLTFK DQFIIAYGGLRGAICFALVFLLPAAVFPRKKLFITAAIVVIFFTVFILGITIRPLVEFLDVKRS NKKQQAVSEEIYCRLFDHVKTGIEDVCGHWGHNFWRDKFKKFDDKYLRKLLIRENQPK S SIVSLYKKLEIKHAIEMAETGMISTVPTF ASLNDCREEKIRKVT S SETDEIRELL SRNL YQ IRQRTLSYNRHSLTADTSERQAKEILIRRRHSLRESIRKDSSLNREHRASTSTSRYLSLPKN TKLPEKLQKRRTISIADGNS SD SD AD AGTT VLNLQPRARRFLPEQF SKK SPQ S YKMEWK NEVDVDSGRDMPSTPPTPHSREKGTQTSGLLQQPLLSKDQSGSEREDSLTEGIPPKPPPRL VWRASEPGSRKARFGSEKP SEQ ID NO: 190. SLC9A3

[0577] MWGLGARGPDRGLLLALALGGLARAGGVEVEPGGAHGESGGFQVVTFEWAHVQDPY VIALWILVASLAKIGFHLSHKVTSVVPESALLIVLGLVLGGIVWAADHIASFTLTPTVFFF YLLPPIVLDAGYFMPNRLFFGNLGTILLYAVVGTVWNAATTGLSLYGVFLSGLMGDLQI GLLDFLLFGSLMAAVDPVAVLAVFEEVHVNEVLFIIVFGESLLNDAVTVVLYNVFESFV ALGGDNVTGVDCVKGIVSFFVVSLGGTLVGVVFAFLLSLVTRFTKHVRIIEPGFVFIISYL SYLTSEMLSLSAILAITFCGICCQKYVKANISEQSATTVRYTMKMLASSAETIIFMFLGISA VNPFIWTWNTAFVLLTLVFISVYRAIGVVLQTWLLNRYRMVQLEPIDQVVLSYGGLRGA VAFALVVLLDGDKVKEKNLFVSTTIIVVFFTVIFQGLTIKPLVQWLKVKRSEHREPRLNE KLHGRAFDHILSAIEDISGQIGHNYLRDKWSHFDRKFLSRVLMRRSAQKSRDRILNVFHE LNLKDAISYVAEGERRGSLAFIRSPSTDNVVNVDFTPRSSTVEASVSYLLRENVSAVCLD MQSLEQRRRSIRDAEDMVTHHTLQQYLYKPRQEYKHLYSRHELTPTEDEKQDREIFHRT MRKRLESFKSTKLGLNQNKKAAKLYKRERAQKRRNSSIPNGKLPMESPAQNFTIKEKDL ELSDTEEPPNYDEEMSGGIEFLASVTKDTASDSPAGIDNPVFSPDEALDRSLLARLPPWLS PGETVVPSQRARTQIPYSPGTFCRLMPFRLSSKSVDSFLQADGPEERPPAALPESTHM SEQ ID NO: 191. SLC9A4

[0578] MALQMFVTYSPWNCLLLLV ALEC SEAS SDLNESANSTAQYASNAWFAAASSEPEEGISV FELDYDYVQIPYEVTLWILLASLAKIGFHLYHRLPGLMPESCLLILVGALVGGIIFGTDHK SPPVMDSSIYFLYLLPPIVLEGGYFMPTRPFFENIGSILWWAVLGALINALGIGLSLYLICQ VKAFGLGDVNLLQNLLFGSLISAVDPVAVLAVFEEARVNEQLYMMIFGEALLNDGITVV LYNMLIAFTKMHKFEDIETVDILAGCARFIVVGLGGVLFGIVFGFISAFITRFTQNISAIEPL IVFMFSYLSYLAAETLYLSGILAITACAVTMKKYVEENVSQTSYTTIKYFMKMLSSVSET LIFIFMGVSTVGKNHEWNWAFICFTLAFCQIWRAISVFALFYISNQFRTFPFSIKDQCIIFYS GVRGAGSFSLAFLLPLSLFPRKKMFVTATLVVIYFTVFIQGITVGPLVRYLDVKKTNKKE SINEELHIRLMDHLKAGIEDVCGHWSHYQVRDKFKKFDHRYLRKILIRKNLPKSSIVSLY KKLEMKQAIEMVETGILS STAF SIPHQAQRIQGIKRLSPEDVESIRDILTSNMYQVRQRTL SYNKYNLKPQTSEKQAKEILIRRQNTLRESMRKGHSLPWGKPAGTKNIRYLSYPYGNPQ S AGRDTRAAGF SDDD S SDPGSP SITF SAC SRIGSLQKQEAQEIIPMKSLHRGRKAF SFGYQ RNTSQEEYLGGVRRVALRPKPLFHAVDEEGESGGESEGKASLVEVRSRWTADHGHGRD HHRSHSPLLQKK SEQ ID NO: 192. SLC9A5 MLRAALSLLALPLAGAAEEPTQKPESPGEPPPGLELFRWQWHEVEAPYLVALWILVASL AKIVFHLSRKVTSLVPESCLLILLGLVLGGIVLAVAKKAEYQLEPGTFFLFLLPPIVLDSGY FMPSRLFFDNLGAILTYAVVGTLWNAFTTGAALWGLQQAGLVAPRVQAGLLDFLLFGS LISAVDPVAVLAVFEEVHVNETLFIIVFGESLLNDAVTVVLYKVCNSFVEMGSANVQAT DYLKGVASLFVVSLGGAAVGLVFAFLLALTTRFTKRVRIIEPLLVFLLAYAAYLTAEMA SLSAILAVTMCGLGCKKYVEANISHKSRTTVKYTMKTLASCAETVIFMLLGISAVDSSK WAWDSGLVLGTLIFILFFRALGVVLQTWVLNQFRLVPLDKIDQVVMSYGGLRGAVAFA LVILLDRTKVPAKDYFVATTIVVVFFTVIVQGLTIKPLVKWLKVKRSEHHKPTLNQELHE HTFDHILAAVEDVVGHHGYHYWRDRWEQFDKKYLSQLLMRRSAYRIRDQIWDVYYRL NIRDAISFVDQGGHVLSSTGLTLPSMPSRNSVAETSVTNLLRESGSGACLDLQVIDTVRS GRDREDAVMHHLLCGGLYKPRRRYKASCSRHFISEDAQERQDKEVFQQNMKRRLESFK STKHNICFTKSKPRPRKTGRRKKDGVANAEATNGKHRGLGFQDTAAVILTVESEEEEEE SD S SETEKEDDEGIIF V ARAT SE VLQEGK V S GSLE VCP SPRIIPP SPTC AEKELP WK S GQGD LAVYVSSETTKIVPVDMQTGWNQSISSLESLASPPCNQAPILTCLPPHPRGTEEPQVPLHL P SDPRS SF AFPP SLAKAGRSRSE SS ADLPQQQELQPLMGHKDHTHL SPGTAT SHWCIQFN RGSRL

[0579] SEQ ID NO: 193. SLC9A6

[0580] MARRGWRRAPLRRGVGSSPRARRLMRPLWLLLAVGVFDWAGASDGGGGEARAMDEE IVSEKQAEESHRQDSANLLIFILLLTLTILTIWLFKHRRARFLHETGLAMIYGLLVGLVLR YGIHVPSDVNNVTLSCEVQSSPTTLLVTFDPEVFFNILLPPIIFYAGYSLKRRHFFRNLGSIL AYAFLGTAISCFVIGSIMYGCVTLMKVTGQLAGDFYFTDCLLFGAIVSATDPVTVLAIFH ELQVDVELYALLFGESVLNDAVAIVLSSSIVAYQPAGDNSHTFDVTAMFKSIGIFLGIFSG SFAMGAATGVVTALVTKFTKLREFQLLETGLFFLMSWSTFLLAEAWGFTGVVAVLFCGI TQAHYTYNNLSTESQHRTKQLFELLNFLAENFIFSYMGLTLFTFQNHVFNPTFVVGAFV AIFLGRAANIYPLSLLLNLGRRSKIGSNFQHMMMFAGLRGAMAFALAIRDTATYARQM MFSTTLLIVFFTVWVFGGGTTAMLSCLHIRVGVDSDQEHLGVPENERRTTKAESAWLFR MWYNFDHNYLKPLLTHSGPPLTTTLPACCGPIARCLTSPQAYENQEQLKDDDSDLILND GDISLTYGD ST VNTEP AT S S APRRFMGNS SED ALDREL AFGDHEL VIRGTRLVLPMDD SE PPLNLLDNTRHGPA

[0581] SEQ ID NO: 194. SLC9A7

[0582] MEPGDAARPGSGRATGAPPPRLLLLPLLLGWGLRVAAAASASSSGAAAEDSSAMEELA TEKEAEESHRQDSVSLLTFILLLTLTILTIWLFKHRRVRFLHETGLAMIYGLIVGVILRYGT PATSGRDKSLSCTQEDRAFSTLLVNVSGKFFEYTLKGEISPGKINSVEQNDMLRKVTFDP EVFFNILLPPIIFHAGYSLKKRHFFRNLGSILAYAFLGTAVSCFIIGNLMYGVVKLMKIMG QLSDKF YYTDCLFFGAIIS ATDPVT VLAIFNELHADVDLYALLFGES VLNDAVAIVLS S SI VAYQPAGLNTHAFDAAAFFKSVGIFLGIFSGSFTMGAVTGVNANVTKFTKLHCFPLLET ALFFLMSWSTFLLAEACGFTGVVAVLFCGITQAHYTYNNLSVESRSRTKQLFEVLHFLA ENFIFSYMGLALFTFQKHVFSPIFIIGAFVAIFLGRAAHIYPLSFFLNLGRRHKIGWNFQHM MMFSGLRGAMAFALAIRDTASYARQMMFTTTLLIVFFTVWIIGGGTTPMLSWLNIRVGV EEP SEEDQNEHHWQ YFRVGVDPDQDPPPNND SFQ VLQGDGPD S ARGNRTKQES AWIFR

[0583] LWYSFDHNYLKPILTHSGPPLTTTLPAWCGLLARCLTSPQVYDNQEPLREEDSDFILTEG DLTLTYGDSTVTANGSSSSHTASTSLEGSRRTKSSSEEVLERDLGMGDQKVSSRGTRLVF PLEDNA

[0584] SEQ ID NO: 195. SLC9A8 MGEKMAEEERFPNTTHEGFNVTLHTTLVVTTKLVLPTPGKPILPVQTGEQAQQEEQSSG

[0585] MTIFFSLLVLAICIILVHLLIRYRLHFLPESVAVVSLGILMGAVIKIIEFKKLANWKEEEMF

[0586] RPNMFFLLLLPPIIFESGYSLHKGNFFQNIGSITLFAVFGTAISAFVVGGGIYFLGQADVISK

[0587] LNMTDSFAFGSLISAVDPVATIAIFNALHVDPVLNMLVFGESILNDAVSIVLTNTAEGLTR

[0588] KNMSDVSGWQTFLQALDYFLKMFFGSAALGTLTGLISALVLKHIDLRKTPSLEFGMMIIF

[0589] AYLPYGLAEGISLSGIMAILFSGIVMSHYTHHNLSPVTQILMQQTLRTVAFLCETCVFAFL

[0590] GLSIFSFPHKFEISFVIWCIVLVLFGRAVNIFPLSYLLNFFRDHKITPKMMFIMWFSGLRGA

[0591] IPYALSLHLDLEPMEKRQLIGTTTIVIVLFTILLLGGSTMPLIRLMDIEDAKAHRRNKKDV

[0592] NLSKTEKMGNTVESEHLSELTEEEYEAHYIRRQDLKGFVWLDAKYLNPFFTRRLTQEDL

[0593] HHGRIQMKTLTNKWYEEVRQGPSGSEDDEQELL

[0594] SEQ ID NO: 196. SLC9A9

[0595] MERQSRVMSEKDEYQFQHQGAVELLVFNFLLILTILTIWLFKNHRFRFLHETGGAMVYG

[0596] LIMGLILRYATAPTDIESGTVYDCVKLTFSPSTLLVNITDQVYEYKYKREISQHNINPHQG

[0597] NAILEKMTFDPEIFFNVLLPPIIFHAGYSLKKRHFFQNLGSILTYAFLGTAISCIVIGLIMYG

[0598] FVKAMIHAGQLKNGDFHFTDCLFFGSLMSATDPVTVLAIFHELHVDPDLYTLLFGESVL

[0599] NDAVAIVLTYSISIYSPKENPNAFDAAAFFQSVGNFLGIFAGSFAMGSAYAIITALLTKFT

[0600] KLCEFPMLETGLFFLLSWSAFLSAEAAGLTGIVAVLFCGVTQAHYTYNNLSSDSKIRTKQ

[0601] LFEFMNFLAENVIFCYMGLALFTFQNHIFNALFILGAFLAIFVARACNIYPLSFLLNLGRK

[0602] QKIPWNFQHMMMFSGLRGAIAFALAIRNTESQPKQMMFTTTLLLVFFTVWVFGGGTTP

[0603] MLTWLQIRVGVDLDENLKEDPSSQHQEANNLDKNMTKAESARLFRMWYSFDHKYLKP

[0604] ILTHSGPPLTTTLPEWCGPISRLLTSPQAYGEQLKEDDVECIVNQDELAINYQEQASSPCS

[0605] PPARLGLDQKASPQTPGKENIYEGDLGLGGYELKLEQTLGQSQLN

Claims

WHAT IS CLAIMED IS:

1. A chimeric polypeptide comprising:(a) an extracellular domain comprising a cytokine-binding domain;(b) an extracellular hinge domain;(c) a transmembrane domain; and(d) an intracellular signaling domain.

2. The chimeric polypeptide of claim 1, wherein the chimeric polypeptide comprises a single polypeptide.

3. The chimeric polypeptide of claim 1, wherein the chimeric polypeptide comprises two polypeptides.

4. The chimeric polypeptide of claim 1, wherein the chimeric polypeptide comprises more than two polypeptides.

5. The chimeric polypeptide of claim 3 or 4, wherein the chimeric polypeptide further comprises a cleaving sequence, wherein the cleaving sequence couples the two or more polypeptides.

6. The chimeric polypeptide of claim 5, wherein the cleaving sequence comprises a P2A, T2A, E2A, F2A, or IRES cleaving sequence.

7. The chimeric polypeptide of any one of claims 1-6, wherein the cytokine-binding domain comprises an scFv.

8. The chimeric polypeptide of any one of claims 1-7, wherein the cytokine-binding domain comprises an IL5R-alpha, IL5R-beta, IFN-gammaRl, IFN-gammaR2, TNFR1, TNFR2 cytokine-binding domain, or any combination thereof.

9. The chimeric polypeptide of any one of claims 1-8, wherein the cytokine-binding domain specifically binds to IL5, IL8, IFN-gamma, TNF-alpha, or any combination thereof.

10. The chimeric polypeptide of claim 9, wherein the cytokine-binding domain specifically binds to IL5, wherein the chimeric polypeptide comprises SEQ ID NO: 93, SEQ ID NO: 16, SEQ ID NO: 84, SEQ ID NO: 17, SEQ ID NO: 85, SEQ ID NO: 73, SEQ ID NO: 78, SEQ ID NO: 12, SEQ ID NO: 15, SEQ ID NO: 21, and SEQ ID NO: 24.

11. The chimeric polypeptide of claim 9, wherein the cytokine-binding domain specifically binds to IL8, wherein the chimeric polypeptide comprises SEQ ID NO: 93, SEQ ID NO: 69, SEQ ID NO: 84, SEQ ID NO: 70, SEQ ID NO: 85, SEQ ID NO: 73, SEQ ID NO: 78, SEQ ID NO: 12, SEQ ID NO: 15, SEQ ID NO: 21, and SEQ ID NO: 24.

12. The chimeric polypeptide of claim 9, wherein the cytokine-binding domain specifically binds to IFN-gamma, wherein the chimeric polypeptide comprises SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, and SEQ ID NO: 33.

13. The chimeric polypeptide of claim 9, wherein the cytokine-binding domain specifically binds to TFN-alpha, wherein the chimeric polypeptide comprises SEQ ID NO: 93, SEQ ID NO: 197, SEQ ID NO: 84, SEQ ID NO: 198, SEQ ID NO: 85, SEQ ID NO: 73, SEQ ID NO: 78, SEQ ID NO: 12, SEQ ID NO: 15, SEQ ID NO: 21, and SEQ ID NO: 24.

14. The chimeric polypeptide of any one of claims 1-13, wherein the cytokine-binding domain is coupled to the intracellular signaling domain, wherein the intracellular signaling domain comprises an IL5R-alpha, ZL5R-beta, IL8R-alpha, IL8R-beta, IFN- gammaRl, IFN-gammaR2, TNFR1, TNFR2, CD8-alpha, CD28 intracellular signaling domain.

15. The chimeric polypeptide of any one of claims 1-14, wherein the hinge domain comprises an IL5R-alpha, IL5R-beta, IFN-gammaRl, IFN-gammaR2, TNFR1, TNFR2, CD8-alpha, CD4, CD28, IgGl, IgG4 hinge domain, one or any combination or all of the four hingedomains of IL8R-alpha, or one or any combination or all of the four hinge domains of IL8R-beta.

16. The chimeric polypeptide of any one of claims 1-15, wherein the transmembrane domain comprises an IL5R-alpha, IL5R-beta, IFN-gammaRl, IFN-gammaR2, TNFR1, TNFR2, CD8-alpha, CD28 transmembrane domain, one or any combination or all of the seven transmembrane domains of IL8R-alpha, or one or any combination or all of the seven transmembrane domains of IL8R-beta.

17. The chimeric polypeptide of any one of claims 1-16, wherein the intracellular signaling domain comprises an IL5R-alpha, IL5R-beta, IFN-gammaRl, IFN-gammaR2, TNFR1, TNFR2 intracellular signaling domain, one or any combination or all of the four intracellular domains of IL8R-alpha, or one or any combination or all of the four intracellular domains of IL8R-beta.

18. A nucleic acid encoding the chimeric polypeptide of any one of claims 1-17.

19. A recombinant vector comprising the nucleic acid of claim 18.

20. The recombinant vector of claim 19, further comprising a promoter operationally linked to the nucleic acid.

21. The recombinant vector of claim 20, wherein the promoter is a constitutive promoter.

22. The recombinant vector of claim 20, wherein the promoter is an inducible promoter.

23. The recombinant vector of claim 20, wherein the promoter comprises an EFl -alpha, CMV, CAG, PgK, or SV40 promoter.

24. The recombinant vector of any one of claims 19-23, wherein the recombinant vector is a viral vector.

25. The recombinant vector of claim 24, wherein the viral vector is a lentiviral vector.

26. An immune cell comprising the nucleic acid of claim 18 or the recombinant vector of any one of claims 19-25.

27. A pharmaceutical composition comprising: any of the chimeric polypeptides of claims 1-17, the nucleic acid of claim 18, the recombinant vector of claims 19-25, or the immune cell of claim 24; and a pharmaceutically acceptable carrier.

28. A method of treating a subject, the method comprising administering to the subject the pharmaceutical composition of claim 27, whereby treating a disease or a condition.

29. The method of claim 28, wherein the disease or condition comprises cancer, autoimmune disease, wound healing, heart disease, or allergy.

30. The method of claim 29, wherein the disease or condition comprises a solid cancer.

31. A method of modulating immune cell migration, the method comprising delivering any of the chimeric polypeptides of claims 1-17, the nucleic acid of claim 18, the recombinant vector of claims 19-25 into an immune cell.

32. The method of claim 31, wherein the modulating comprises increasing immune cell migration.

33. The method of claim 31 or 32, wherein the immune cell is a human immune cell.

34. The method of claim 33, wherein the immune cell comprises a T cell, B cell, Natural Killer (NK) cell, dendritic cell (DC), macrophage, monocyte, eosinophil, basophil,neutrophil, mast cell, lymphoid progenitor cell, myeloid progenitor cell, hematopoietic stem cell, red blood cell, platelet, or megakaryocyte.

35. The method of claim 34, wherein the immune cell comprises a T cell.

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