Activated hepatocyte growth factor

Abstract

The present disclosure relates to activated hepatocyte growth factor (HGF) and compositions and methods for producing the same, including, for example, cells and / or populations of cells engineered to co-express hepsin and HGF. In some embodiments, it includes cell(s) comprising one or more polynucleotide sequence) s) encoding a hepatocyte growth factor (HGF) polypeptide or biologically active variant thereof operably linked to an exogenous promoter; and one or more polynucleotide sequence) s) encoding a hepsin polypeptide or enzymatically active variant thereof operably linked to an exogenous promoter.

Description

Attorney Docket No.: 49082-0007WO1 ACTIVATED HEPATOCYTE GROWTH FACTOR CROSS-REFERENCE TO RELATED APPLICATIONS This application is an International Application which claims priority to U.S. Patent Application No. 63 / 678,075, filed August 1, 2024. The disclosure of the foregoing application is hereby incorporated by reference in its entirety. SEQUENCE LISTING This application contains a Sequence Listing that has been submitted electronically as an XML file named “49082-0007WO1_SL_ST26.XML.” The XML file, created on July 29, 2025, is 76,872 bytes in size. The material in the XML file is hereby incorporated by reference in its entirety. TECHNICAL FIELD The present disclosure relates to hepsin activated hepatocyte growth factor and compositions and methods for producing the same. BACKGROUND There is an ongoing need for improvements in production of protease activated proteins such as hepatocyte growth factor (HGF). The compositions and methods described herein have been developed toward this end. SUMMARY Hepatocyte Growth Factor (HGF) has been investigated for the treatment of corneal haze of scarring. See, US Pat. No. 10,449,234. Other eye injuries and diseases have been identified that could benefit from new treatments. However, current methods for production of activated HGF produce low yields and require, for example, bovine serum albumin for proteolytic cleavage of pro-HGF to its activated form and for growth of the cells. A need exists for improved production methods, including, for example, process for producing active HGF without the need for additional enzyme addition or processing, e.g., a serum-free process.Attorney Docket No.: 49082-0007WO1 Thus, provided herein, among other things, are compositions and methods for protease activated protein production, e.g., hepsin activated HGF production. Provided herein are cell(s) engineered to express a hepatocyte growth factor (HGF) polypeptide or biologically active variant thereof and a hepsin polypeptide or enzymatically active variant thereof. Also provided herein are cell(s) comprising one or more polynucleotide sequence(s) encoding a hepatocyte growth factor (HGF) polypeptide or biologically active variant thereof operably linked to an exogenous promoter; and one or more polynucleotide sequence(s) encoding a hepsin polypeptide or enzymatically active variant thereof operably linked to an exogenous promoter. Also provided herein are population(s) of cells engineered to express a hepatocyte growth factor (HGF) polypeptide or biologically active variant thereof and a hepsin polypeptide or enzymatically active variant thereof. Also provided herein are population(s) of cells comprising one or more polynucleotide sequence(s) encoding a hepatocyte growth factor (HGF) polypeptide or biologically active variant thereof operably linked to an exogenous promoter; and one or more polynucleotide sequence(s) encoding a hepsin polypeptide or enzymatically active variant thereof operably linked to an exogenous promoter. In some embodiments, the promoter is selected from the group consisting of human CMV, simian CMV, EF1a, SV40, human PGK1, mouse PGK1, Ubc, Human beta actin, CAG, TRE, UAS, CaMKIIa, H1, RSV, UB, and U6. In some embodiments, the promoter is a cytomegalovirus immediate-early enhancer / promoter. In some embodiments, the HGF polypeptide or biologically active variant thereof comprises a hepsin cleavage amino acid motif. In some embodiments, the hepsin cleavage amino acid motif is selected from the group consisting of KKNR, KKTR, KKLR, KQNR, KQTR, KQLR, PQNR, PQTR, PQLR, PKNR, PKTR and PKLR. In some embodiments, the HGF polypeptide or biologically active variant thereof comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 32. In some embodiments, the HGF polypeptide or biologically active variant thereof comprises a signal sequence. In some embodiments, the signal sequence is a native HGF signal sequence. In some embodiments, the signal sequence is not a native HGF signal sequence. In some embodiments, the signal sequence is selected from the group consisting of SEQ ID NOs:40–57. In some embodiments, the hepsin polypeptide or enzymatically active variant thereof comprises an amino sequence having at least 80% identity to SEQ ID NO: 29.Attorney Docket No.: 49082-0007WO1 In some embodiments, the hepsin polypeptide or enzymatically active variant thereof comprises a signal sequence. In some embodiments, the signal sequence is selected from the group consisting of SEQ ID NOS:40–57. In some embodiments, the cell is a mammalian cell. In some embodiments, the cell is a CHO cell. In some embodiments, the population of cells comprises or consists of mammalian cells. In some embodiments, the population of cells comprises or consists of CHO cells. In some embodiments, the cell comprises polynucleotide sequence(s) encoding a ratio of from 1:1 to 1:1000 copies of the hepsin polypeptide or enzymatically active variant thereof to the HGF polypeptide or biologically active variant thereof. In some embodiments, the ratio is at least 50 copies of the HGF polypeptide or enzymatically active variant thereof to 1 of the hepsin polypeptide or biologically active variant thereof. In some embodiments, the ratio is from 1:50 to 1:1000. In some embodiments, the ratio is from 1:50 to 1:100. In some embodiments, the ratio is from 1:79 to 1:127, optionally about 1:100. In some embodiments, the population of cells cell comprises polynucleotide sequence(s) encoding a ratio of from 1:1 to 1:1000 copies of the hepsin polypeptide or enzymatically active variant thereof to the HGF polypeptide or biologically active variant thereof. In some embodiments, the ratio is at least 50 copies of the HGF polypeptide or enzymatically active variant thereof to 1 of the hepsin polypeptide or biologically active variant thereof. In some embodiments, the ratio is about 1:100. Also provided herein are compositions comprising: a first nucleic acid expression construct comprising a polynucleotide sequence encoding hepatocyte growth factor (HGF) or a biologically active variant thereof; and a second nucleic acid expression construct comprising a polynucleotide sequence encoding hepsin or an enzymatically active variant thereof. In some embodiments, the HGF polypeptide or biologically active variant thereof comprises a hepsin cleavage amino acid motif. In some embodiments, the hepsin cleavage amino acid motif is selected from the group consisting of KKNR, KKTR, KKLR, KQNR, KQTR, KQLR, PQNR, PQTR, PQLR, PKNR, PKTR and PKLR. In some embodiments, the HGF polypeptide or biologically active variant thereof comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 32. In some embodiments, the HGF polypeptide or biologically active variant thereof comprises a signal sequence. In some embodiments, the signal sequence is a native HGF signal sequence. In some embodiments,Attorney Docket No.: 49082-0007WO1 the signal sequence is not a native HGF signal sequence. In some embodiments, the signal sequence is selected from the group consisting of SEQ ID NOs:40–57. In some embodiments, the hepsin polypeptide or enzymatically active variant thereof comprises an amino sequence having at least 80% identity to SEQ ID NO: 29. In some embodiments, the hepsin polypeptide or enzymatically active variant thereof comprises a signal sequence. In some embodiments, the signal sequence is selected from the group consisting of SEQ ID NOS: 40–57. In some embodiments, the first nucleic acid expression construct further comprises a nucleic acid sequence encoding a promoter operably linked to the polynucleotide encoding HGF. In some embodiments, the promoter is selected from the group consisting of human CMV, simian CMV, EF1a, SV40, human PGK1, mouse PGK1, Ubc, Human beta actin, CAG, TRE, UAS, CaMKIIa, H1, RSV, UB, and U6. In some embodiments, the promoter is a cytomegalovirus immediate-early enhancer / promoter. In some embodiments, the second nucleic acid expression construct further comprises a nucleic acid sequence encoding a promoter, operably linked to the polynucleotide encoding hepsin or an enzymatically active variant thereof. In some embodiments, the promoter is selected from the group consisting of human CMV, simian CMV, EF1a, SV40, human PGK1, mouse PGK1, Ubc, Human beta actin, CAG, TRE, UAS, CaMKIIa, H1, RSV, UB, and U6. In some embodiments, the promoter is a cytomegalovirus immediate-early enhancer / promoter. In some embodiments, the first and / or second nucleic acid expression construct(s) each, independently, further comprise one or more of: a nucleic acid sequence encoding an attachment site, a nucleic acid sequence encoding a selectable marker, a nucleic acid sequence encoding a posttranscriptional regulatory element, and a poly A signal sequence. Also provided herein are cell(s) comprising a composition described herein. Also provided herein are cell(s) transformed, transfected, or transduced with a composition described herein. Also provided herein are population(s) of cells transformed, transfected, or transduced with a composition described herein. In some embodiments, the cell or population of cells is(s) are transformed, transfected, or transduced with a plurality of first and second nucleic acid expression constructs in a ratio of from 1:1 to 1:1000 of the second (hepsin) construct to the first (HGF) construct. In some embodiments, the cell or population of cells is(s) are transformed, transfected, or transduced with a plurality of first and second nucleic acid expression constructs in a ratio of from 1:10 to 1:1000 of the second (hepsin) construct to the first (HGF)Attorney Docket No.: 49082-0007WO1 construct. In some embodiments, the cell or population of cells is transformed, transfected, or transduced with a plurality of first and second nucleic acid constructs in a ratio of about 1:100. In some embodiments, the cell(s) are mammalian cell. In some embodiments, the cell(s) are CHO cell(s). Also provided herein are methods of producing activated HGF, comprising: culturing a cell described herein or a population of cells described herein under conditions suitable for a) expression of the HGF or biologically active variant thereof and of the hepsin or enzymatically active portion thereof; and b) cleavage of the HGF or biologically active variant thereof with the hepsin or enzymatically active portion thereof, thereby producing activated HGF. In some embodiments the method further comprises: isolating and / or purifying the HGF or biologically active variant thereof. In some embodiments, the activated HGF comprises a first polypeptide comprising an amino acid sequence having at least 80% identity to SEQ ID NO:33 and a second polypeptide comprising an amino acid sequence having at least 80% identity to SEQ ID NO:34. In some embodiments, the first polypeptide and second polypeptide are linked by one or more disulfide bonds. In some embodiments, the activated HGF is capable of binding to c-MET and / or activating the MAPK pathway in epithelial cells. Also provided herein are activated HGF polypeptide(s) produced according to a method described herein. Also provided herein are methods of producing a cell line co-expressing HGF and hepsin, comprising: transforming, transducing, or transfecting a population of cells with a composition described herein. In some embodiments, the population of cells is transformed, transfected, or transduced with a plurality of first and second nucleic acid expression constructs in a ratio of 1:1 to 1:1000 of the second (hepsin) construct to the first (HGF) construct. In some embodiments, the population of cells is transformed, transfected, or transduced with a plurality of first and second nucleic acid expression constructs in a ratio of 1:10 to 1:1000 of the second (hepsin) construct to the first (HGF) construct. In some embodiments, the population of cells is transformed, transfected, or transduced with a plurality of first and second nucleic acid expression constructs in a ratio of about 1:100 of the second (hepsin) construct to the first (HGF) construct.Attorney Docket No.: 49082-0007WO1 In some embodiments, the cells are mammalian cell. In some embodiments, the cell(s) are CHO cells. In some embodiments the method further comprises: isolating a cell from the transformed, transduced, or transfected population of cells. In some embodiments, the isolated cell comprises polynucleotide sequence(s) encoding a ratio of from 1:1 to 1:1000 copies of the hepsin polypeptide or enzymatically active variant thereof to the HGF polypeptide or biologically active variant thereof. In some embodiments, the ratio is at least 50 copies of the HGF polypeptide or enzymatically active variant thereof to 1 of the hepsin polypeptide or biologically active variant thereof. In some embodiments, the ratio is from 1:50 to 1:1000. In some embodiments, the ratio is from 1:50 to 1:100. In some embodiments, the ratio is from 1:79 to 1:127, optionally about 1:100. Throughout this application, various embodiments may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the disclosure. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range. As used in the specification and claims, the singular forms “a”, “an” and “the” include plural references unless the context clearly dictates otherwise. For example, the term “a sample” includes a plurality of samples, including mixtures thereof. The terms “determining,” “measuring,” “evaluating,” “assessing,” “assaying,” and “analyzing” are often used interchangeably herein to refer to forms of measurement. The terms include determining if an element is present or not (for example, detection). These terms can include quantitative, qualitative or quantitative and qualitative determinations. Assessing can be relative or absolute. “Detecting the presence of” can include determining the amount of something present in addition to determining whether it is present or absent depending on the context. As used herein, the term “about” a number refers to that number plus or minus 10% of that number. The term “about” a range refers to that range minus 10% of its lowest value and plus 10% of its greatest value.Attorney Docket No.: 49082-0007WO1 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 belongs. Methods and materials are described herein for use in the present invention; other, suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. Other features and advantages of the invention will be apparent from the following detailed description and figures, and from the claims. DESCRIPTION OF DRAWINGS The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee. FIG. 1 is a map of plasmid pFCS-hHGF-WPRE-SIN. FIG. 2 shows sequences of the transgene insert sequence for plasmid pFCS-hHGF- WPRE-SIN (nucleotide: SEQ ID NO. 35; amino acid: SEQ ID NO: 1). FIG. 3 is a map of plasmid pFCS-newhHepsin-WPRE_SIN. FIG. 4 shows sequences of the transgene insert sequence for plasmid pFCS- newhHepsin-WPRE_SIN (nucleotide: SEQ ID NO: 36; amino acid: SEQ ID NO: 37). FIG. 5 is a map of plasmid pFCS-SPhHGF-WPRE-SIN. FIG. 6 shows sequences of the transgene insert sequence for plasmid pFCS-SPhHGF- WPRE-SIN (nucleotide: SEQ ID NO: 38; amino acid: SEQ ID NO: 39). FIG. 7 is a map of plasmid 207attB-GS-Hepsin-WPRE. FIG. 8 shows sequences of the transgene insert sequence for plasmid 207attB-GS- Hepsin-WPRE (nucleotide: SEQ ID NO: 36; amino acid: SEQ ID NO: 37). FIG. 9 is a map of plasmid 207attB-GS-hHGF-WPRE. FIG. 10 shows sequences of the transgene insert sequence for plasmid 207attB-GS- hHGF-WPRE (nucleotide: SEQ ID NO: 35; amino acid: SEQ ID NO: 1). FIGS. 11A–11D show cell growth (FIG. 11A and FIG. 11C) and viability (FIG. 11B and FIG. 11D) curves for pooled cell cultures transduced with pFCS-hHGF-WPRE-SIN (FIG. 11A and FIG. 11B) and pFCS-SPhHGF-WPRE-SIN (FIG. 11C and FIG. 11D).Attorney Docket No.: 49082-0007WO1 FIG. 12 shows a non-reduced SDS-PAGE of HGF produced by pooled cell cultures transduced with pFCS-hHGF-WPRE-SIN or pFCS-SPhHGF-WPRE-SIN. Lane 1: Molecular weight standard; Lane 2: blank; Lane 3: pFCS-hHGF-WPRE-SIN 4d; Lane 4: pFCS-hHGF- WPRE-SIN 8d; Lane 5: pFCS-hHGF-WPRE-SIN 12d; Lane 6: pFCS-SPhHGF-WPRE-SIN 4d; Lane 7: pFCS-SPhHGF-WPRE-SIN 8d; Lane 8: pFCS-SPhHGF-WPRE-SIN 10d; Lane 9: HGF standard (0.2 μg load); Lane 10: HGF standard (2 μg load); Lane 11: HGF standard (5 μg load). FIG. 13 shows a reduced SDS-PAGE of HGF produced by pooled cell cultures transduced with pFCS-hHGF-WPRE-SIN or pFCS-SPhHGF-WPRE-SIN. Lane 1: Molecular weight standard; Lane 2: blank; Lane 3: pFCS-hHGF-WPRE-SIN 4d; Lane 4: pFCS-hHGF- WPRE-SIN 8d; Lane 5: pFCS-hHGF-WPRE-SIN 12d; Lane 6: pFCS-SPhHGF-WPRE-SIN 4d; Lane 7: pFCS-SPhHGF-WPRE-SIN 8d; Lane 8: pFCS-SPhHGF-WPRE-SIN 10d; Lane 9: HGF standard (0.2 μg load); Lane 10: HGF standard (2 μg load); Lane 11: HGF standard (5 μg load). FIGS. 14A–14B show non-reduced (FIG. 14A) and reduced (FIG. 14B) SDS-PAGE gels of cell pool samples from pools containing and not containing hepsin, as well as a control HGF standard. Lane 1: Molecular weight standard; Lane 2: conditioned media: Lane 3 HGF std (2 μg load); Lane 4: pFCS-hHGF-WPRE-SIN 3x HGF transductions / 2x hepsin transduction, initial protein; Lane 5: pFCS-hHGF-WPRE-SIN 3x HGF transduction / 3x hepsin, initial protein; Lane 6: pFCS-hHGF-WPRE-SIN 3x, productivity culture media 9d; Lane 7: 3x HGF transduction / 1x hepsin production, productivity culture media 9d; Lane 8: pFCS-hHGF-WPRE-SIN 3x HGF transduction / 2x hepsin transduction, productivity culture media 9d; Lane 9: Ladder (0.2 μg load). FIG. 15 shows cell viability curves after glutamine selection for cells transduced with HGF and hepsin at HGF:hepsin ratios (from left to right), 100:1, 10:1, and 1:1. FIGS. 16A–16B show non-reduced (FIG. 16A) and reduced (FIG. 16B) SDS-PAGE gels of samples GPExTMLightning cell pool samples from the pool lacking hepsin and the pool produced using the 100:1 ratio of HGF:Hepsin, as well as a control HGF standard. Lane 1: ladder; Lane 2: HGF Standard (2 μg load); Lane 3: Conditioned Media (Fed Batch +Q); Lane 4: Line 1-1; Lane 5: Line 1-2; Lane 6: Line 1-3; Lane 7: Line 4-1 (100:1); Lane 8: Line 4-2 (100:1); Lane 9: Line 4-3 (100:1). FIGS. 17A–17B show viable cell density (FIG. 17A) and cell viability (FIG. 17B) through the culture for a fed-batch productivity study of Line 1 (squares) and Line 4 (diamonds).Attorney Docket No.: 49082-0007WO1 FIGS. 18A–18D show non-reduced (FIG.18A and FIG. 18B) and reduced (FIG. 18C and FIG. 18D) SDS-PAGE gels of samples from Line 1 and Line 4 cell pools harvested on various days throughout a production run. FIG. 18A and FIG. 18C: Lane 1: ladder; Lane 2: HGF standard (2 μg load); Lane 3: Conditioned Fed Batch Media (+Q); Lane 4: Fresh Fed Batch Media (-Q); Lane 5: Line 1 d8 (7.5 μl load); Lane 6: Line 2 d8 (7.5 μl load); Lane 7: Line 1 d10 (7.5 μl load); Lane 8: Line 2 d10 (7.5 μl load); Lane 9: Line 1 d8 (1.5 μl load); Lane 10: Line 2 d8 (1.5 μl load); Lane 11: Line 1 d10 (1.5 μ load); Lane 12: Line 2 d10 (1.5 μ load). FIG. 18B and FIG. 18D: Lane 1: ladder; Lane 2: HGF standard (2 μg load); Lane 3: Conditioned Fed Batch Media (+Q); Lane 4: Fresh Fed Batch Media (-Q); Lane 5: Line 1 d12 (1.3 μl load); Lane 6: Line 2 d12 (1.3 μl load); Lane 7: Line 1 d16 (1.3 μl load); Lane 8: Line 2 d16 (1.3 μl load); Lane 9: Line 1 d17 harvest (1.3 μl load); Lane 10: Line 2 d17 harvest (1.3 μl load); Lane 11: HGF standard (1 μg load). FIG. 19 shows non-reduced (left) and reduced (right) SDS-PAGE gels of material purified from cell pools for Lines 1 (middle lane in each) and 4 (right lane in each). Left lane: molecular weight standard. DETAILED DESCRIPTION Provided herein are, among other things, cell(s) engineered to co-express hepsin and HGF, e.g., activated HGF; HGF, e.g., activated HGF produced by such cells; and compositions and methods for producing the same. HEPSIN Hepsin (e.g., UNiProtKB P05981; SEQ ID NO:28) is a type II transmembrane serine protease that cleaves extracellular substrates, and contributes to the proteolytic processing of growth factors, such as HGF and MST1 / HGFL. See, e.g., Ganesan et al., “Proteolytic Activation of Pro-Macrophage-Stimulating Protein by Hepsin” Mol. Cancer Res. 9:1175–86 (2011); Herter et al., “Hepatocyte Growth Factor is a Preferred In Vitro substrate for Human hepsin, a Membrane-Anchored Serine Protease Implicated in Prostate and Ovarian Cancers,” Biochem. J. 390:125–36 (2005). It has cleavage activity after basic amino-acid residues (Arg, His, Lys), with Arg strongly preferred to Lys. Hepsin exhibits strong P1–P4 substrate specificity at the P1 position for arginine over lysine, and favors threonine, leucine or asparagine at the P2, glutamine or lysine at the P3, and proline or lysine at the P4 position (with P4 to P1 being oriented N-terminally to C-terminally relative to the cut site as follows:Attorney Docket No.: 49082-0007WO1 P4-P3-P2-P1↓). Ganesan et al., “Proteolytic Activation of Pro-Macrophage-Stimulating Protein by Hepsin” Mol. Cancer Res. 9:1175–86 (2011). The hepsin protein comprises a cytoplasmic domain (amino acid residues 1–23 of SEQ ID NO:28), a helical transmembrane domain (amino acid residues 24–44 of SEQ ID NO:28), and an extracellular domain (SEQ ID NO:29, corresponding to amino acid residues 45–417 of SEQ ID NO:28). The extracellular region of hepsin is synthesized as a single chain, though a proteolytic activation step cleaves the peptide bond between Arg162 and Ile163, separating the serine protease domain (the 255-residue C-terminal section, SEQ ID NO:30) from the rest of the protein. The two chains remain covalently bound to each other through a disulfide bond. The protease’s catalytic triad is comprised of the amino acids correspond to H203, D257, and S353 in SEQ ID NO:28. Somoza et al., “The Structure of the Extracellular Region of Human Hepsin Reveals a Serine Protease Domain and a Novel Scavenger Receptor Cysteine-Rich (SRCR) Domain,” Structure 11:1123–31 (2003). In some cases, hepsin comprises one or more of the post-translational modifications (with positions correspond to those of SEQ ID NO:28) as shown in the table below:Thus, in some cases, the amino acid sequence encoding the hepsin or enzymatically active variant thereof comprises cysteine residues at one or more of the following pairs of amino acids, numbered relative to SEQ ID NO:28: C77 and C140, C90 and C150, C119 and C138, C153 and C277, C188 and C204, C291 and C359, C322 and C338, C349 and C381. In some cases, the amino acid sequence encoding the hepsin or enzymatically active variant thereof comprises an asparagine residue at the position corresponding to 112 of SEQ ID NO:28.Attorney Docket No.: 49082-0007WO1 In some embodiments, the hepsin is a polypeptide comprising or consisting of SEQ ID NO:28 or an enzymatically active variant thereof. In some embodiments, the enzymatically active variant thereof comprises or consists of SEQ ID NO:29. In some embodiments, the enzymatically active variant thereof comprises or consists of SEQ ID NO:30. In some embodiments, the hepsin is polypeptide comprising or consisting of a polypeptide having at least 99%, e.g., at least 98%, 97%, 96%, 95%, 90%, 85%, or 80% sequence identity to SEQ ID NO:28. In some embodiments, the hepsin is polypeptide comprising or consisting of a polypeptide having at least 99%, e.g., at least 98%, 97%, 96%, 95%, 90%, 85%, or 80% sequence identity to an enzymatically active variant of SEQ ID NO:28 (e.g., SEQ ID NO:29 or SEQ ID NO:30). HEPATOCYTE GROWTH FACTOR Hepatocyte growth factor (HGF, e.g., UniProt ID No. P14210) is a cMet kinase agonist that, inter alia, stimulates epithelial cell proliferation, motility, morphogenesis, and angiogenesis in various organs via the tyrosine kinase signaling pathway, and plays major roles in embryonic organ development and adult organ regeneration and wound healing. HGF is an approximately 84 kDa protein comprising of two subunits: an α-subunit of an apparent molecular weight of 69 kDa and a β-subunit of an apparent molecular weight of 34 kDa linked by a single disulfide bond. HGF is produced as a pro-HGF molecule of 728 amino acids by mesenchymal stromal cells (e.g. fibroblasts and macrophages) wherein the first 1-31 amino acid residues correspond to a secretion signaling sequence (Matsumoto and Nakamura, in Encyclopedia of Endocrine Diseases, Elsevier, 2004, 436-442). The primary amino acid sequence of Pro- HGF is organized into a domain structure of four Kringle (K) domains (Sigurdardottir, et al., 2015, Chem. Sci., 6:6147-6157). As used herein, a “Kringle” domain refers to a polypeptide linear sequence folded into triple-looped, disulfide cross-linked domains (Simonneau, et al., 2015, Chem. Sci., 6:2110-2121). Kringle domains are present in a variety of polypeptides including apolipoprotein A, blood coagulation factor XII, plasminogen, and HGF. The name “Kringle” is derived from the Scandinavian pastry that these structures resemble. Kringle domains are believed to mediate binding interactions between proteins, membranes, and phospholipids. Upon cleavage of the secretion signaling sequence, proteolytic cleavage at a trypsin- like site (R494-V495) between the fourth Kringle and the C-terminal serine-proteinaseAttorney Docket No.: 49082-0007WO1 homology (SPH) domain of the pro-HGF yields the mature, activated HGF. A trypsin-like cleavage site is a peptide bond following a positively-charged amino acid (e.g. lysine or arginine). Upon activation, HGF binds to and activates its receptor, cMet (also known as the MET tyrosine kinase), which leads to tyrosine phosphorylation, further effecting the recruitment of a variety of downstream adaptor molecules and modulation of various intracellular pathways and biological activities collectively known as the invasive growth program (Nakamura, T., 1991, Prog. Growth Factor Res., 3:67-85; Bottaro, et al., 1991, Science 251:802-804). Six naturally occurring isoforms of HGF resulting from alternative splicing of the HGF mRNA gene transcripts are known in the literature, among which, the major isoforms are isoforms 1 (e.g., SEQ ID NO:2) and 2 (e.g., SEQ ID NO:3) (Bottaro, et al., 1991, Science, 251:802-804; Chan et al., 1991, Science, 254:1382-1385; Lokker and Godowski, 1992, EMBO J., 11:2503-2510; Cioce, et al., 1996, J. Biol. Chem., 271:13110-13115). As used herein, the term “isoform” refers to proteins that result from alternative splicing of the pre- mRNA encoding HGF. The isoform 1 mRNA transcript encodes the longest HGF gene sequence, which comprises 728 amino acid residues. The second major HGF isoform is encoded by the mRNA transcript of isoform 2, which lacks multiple 3’ exons but includes an alternate 3’ exon relative to the isoform 1 transcript. The HGF protein encoded by the isoform 2 mRNA transcript comprises 290 amino acid residues, and is truncated after the second Kringle domain as compared to the isoform 1 HGF protein (Miyazawa, et al., 1991, Eur. J. Biochem., 197:15-22). The isoform 3 mRNA transcript lacks an in-frame coding segment present in isoform 1, and encodes an HGF protein comprising 723 amino acid residues (e.g., SEQ ID NO:1) lacking the sequence “SFLPS” at positions 161-165 within the first Kringle domain of isoform 1 (Rubin et al., 1991, Proc. Natl. Acad. Sci., 88:415-419). The isoform 4 HGF protein (e.g., SEQ ID NO:4) is a smaller molecule comprising residues 1- 296, wherein the amino acid sequence extends only through the second Kringle domain (Chan et al., 1991, Science, 254:1382-1385). The isoform 5 HGF protein (e.g., SEQ ID NO:5) is similar to the isoform 2 HGF protein with the additional deletion of residues 161- 165 (SFLPS) as in isoform 3 HGF (Rubin et al., 1991, Proc. Natl. Acad. Sci., 88: 415-419). The isoform 6 HGF protein (also known as NK1) (e.g., SEQ ID NO:6) is the smallest of all HGF isoforms, comprising only 210 amino acids (Cioce, et al., 1996, J. Bio. Chem., 271:13110-13115). Both the full length and truncated isoforms of HGF have been shown to bind to cMet, albeit with different potency and interactions with heparin sulfate proteoglycan of theAttorney Docket No.: 49082-0007WO1 extracellular matrix, which help prolong the circulating half-lives of HGF in vivo (Masumoto and Yamamoto, 1991, Biochem. Biophys. Res. Commun., 174:90-95; Montesano, et al., 1998, Cell Growth Differ., 9:355-365; Sakata et al., 1997, J. Biol. Chem., 272:9457-9463; Stahl et al., 1997, Biochem. J., 326:763-772). While isoforms 1 and 3 require the proteolytic cleavage at R494-V495 to become biologically active, the truncated isoforms 2, 4, 5 and 6 do not possess the R494-V495 cleavage site and as such, this activation step is not required for their biological activities. However, the truncated isoforms 2, 4, 5 and 6 are known to be generally less potent than the full length isoforms 1 and 3 counterparts (Stahl et al., 1997; Montesano, et al., 1998; supra). In some embodiments, the HGF is a polypeptide comprising the amino acid sequence of any one of SEQ ID NOS: 1, 2, and 7-27, with or without the signal sequence (i.e., the amino acids corresponding to amino acids 1–31 of SEQ ID NO: 1, 2, and 7–27). In some embodiments, the HGF is a polypeptide comprising the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2, with or without the signal sequence (e.g., the amino acids corresponding to amino acids 1–31 of SEQ ID NO: 1 or SEQ ID NO: 2). In some embodiments, the HGF is a polypeptide comprising the amino acid sequence of SEQ ID NO: 1, with or without the signal sequence (e.g.,, the amino acids corresponding to amino acids 1–31 of SEQ ID NO: 1). In some embodiments, the HGF is a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, with or without the signal sequence (e.g., the amino acids corresponding to amino acids 1–31 of SEQ ID NO: 2). In some embodiments, the HGF is a polypeptide comprising an activated form the amino acid sequence of SEQ ID NO. 1 or SEQ ID NO. 2. The term “activated form” refers to, inter alia, an HGF polypeptide from which the signal sequence (e.g., amino acids 1–31 of SEQ ID NO: 1 or SEQ ID NO: 2) has been cleaved and that has been cleaved between R489 and V490 of SEQ ID NO:1 or between R494 and V495 of SEQ ID NO:2 to form the disulfide-linked alpha and beta chains of HGF. In some embodiments, the N-terminal amino acid (e.g., amino acid 32 of SEQ ID NO: 1 or SEQ ID NO: 2) is pyrrolidone carboxylic acid, e.g., resulting in cleavage of the signal sequence. In some embodiments, polypeptides of the invention exist in the HGF precursor (pro-HGF) form wherein the peptide bond between R489 and V490 of SEQ ID NO:1 or R494 and V495 of SEQ ID NO:2 is intact. In some embodiments, polypeptides of the invention can be converted to an active form by proteolytic cleavage of a trypsin-like cleavage site between the R489 and V490 amino acid residues of SEQ ID NO:1 or between the R494 and V495 amino acid residues of SEQ ID NO:2 pro-HGF protein in vivo. In some embodiments, polypeptides of the invention can be converted to anAttorney Docket No.: 49082-0007WO1 active form by proteolytic cleavage of a trypsin-like cleavage site between the R489 and V490 amino acid residues of SEQ ID NO:1 or between the R494 and V495 amino acid residues of SEQ ID NO:2 pro-HGF protein in vitro. In some embodiments, the HGF is a polypeptide comprising the amino acid sequence of amino acids 32-723 of SEQ ID NO:1 pro-HGF protein. In some embodiments, the HGF is a polypeptide comprising the amino acid sequence of amino acids 32-723 of SEQ ID NO:1 activated protein. In some embodiments, the HGF is a polypeptide comprising the amino acid sequence of amino acids 32-728 of SEQ ID NO:2 pro-HGF protein. In some embodiments, the HGF is a polypeptide comprising the amino acid sequence of amino acids 32-728 of SEQ ID NO:2 activated protein. In some embodiments, the HGF is pro-HGF protein comprising a hepsin cleavage amino acid motif selected from the group consisting of NR, TR, and LR. In some cases, when cleaved at the hepsin cleavage amino acid motif, the pro-HGF protein is converted to an active form. In some cases, the HGF is a pro-HGF protein comprising a single hepsin cleavage amino acid motif selected from the group consisting of NR, TR, and LR. In some cases, when cleaved at the hepsin cleavage amino acid motif, the pro-HGF protein is converted to an active form. In some embodiments, the HGF is pro-HGF protein comprising a hepsin cleavage amino acid motif selected from the group consisting of KKNR, KKTR, KKLR, KQNR, KQTR, KQLR, PQNR, PQTR, PQLR, PKNR, PKTR and PKLR. In some cases, when cleaved at the hepsin cleavage amino acid motif, the pro-HGF protein is converted to an active form. In some embodiments, the HGF is pro-HGF protein comprising a single hepsin cleavage amino acid motif selected from the group consisting of KKNR, KKTR, KKLR, KQNR, KQTR, KQLR, PQNR, PQTR, PQLR, PKNR, PKTR and PKLR. In some cases, when cleaved at the hepsin cleavage amino acid motif, the pro-HGF protein is converted to an active form. In some embodiments, the HGF is a pro-HGF protein comprising a RV dipeptide. In some cases, when cleaved at the RV dipeptide, the pro-HGF is converted to an active form. In some embodiments, the HGF is a pro-HGF protein comprising a single RV dipeptide. In some cases, when cleaved at the RV dipeptide, the pro-HGF is converted to an active form.Attorney Docket No.: 49082-0007WO1 In some embodiments, the HGF is a polypeptide variant of SEQ ID NO: 1 or SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, or SEQ ID NO: 6, with or without the signal sequence (e.g., the amino acids corresponding to amino acids 1–31 of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, or SEQ ID NO: 6). As used herein, the term “variant” is meant to indicate a polypeptide differing from another polypeptide by one or more amino acid substitutions, deletions or insertions resulting from mutations in the nucleic acid coding for the polypeptide. In some embodiments, the HGF is a polypeptide comprising an amino acid sequence having at least 80%, e.g., at least 90% sequence identity to SEQ ID NO:1 or SEQ ID NO: 2, with or without the signal sequence (e.g., the amino acids corresponding to amino acids 1–31 of SEQ ID NO: 1, 2). In some embodiments, the HGF is a polypeptide comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO:1, SEQ ID NO: 2, with or without the signal sequence (e.g., the amino acids corresponding to amino acids 1–31 of SEQ ID NO: 1, 2). In some embodiments, the HGF is a polypeptide comprising an amino acid sequence having at least 98% sequence identity to SEQ ID NO:1, SEQ ID NO: 2, with or without the signal sequence (e.g., the amino acids corresponding to amino acids 1–31 of SEQ ID NO: 1, 2). In some embodiments, the HGF is a polypeptide comprising an amino acid sequence having at least 99% sequence identity to SEQ ID NO:1, SEQ ID NO: 2, with or without the signal sequence (e.g., the amino acids corresponding to amino acids 1–31 of SEQ ID NO: 1, 2). In some embodiments, the HGF variant can (1) bind to c-MET and / or (2) activate the MAPK pathway in epithelial cells (e.g., human corneal epithelial cells). In some embodiments, the HGF is a polypeptide comprising a mutation in an amino acid sequence of wild-type HGF isoform 1 (e.g., SEQ ID NO: 2, with or without the signal sequence (e.g., the amino acids corresponding to amino acids 1–31 of SEQ ID NO: 2) at one or more of positions 62, 64, 77, 95, 125, 127, 130, 132, 137, 142, 148, 154, 170, 173 and 193. In some embodiments, analogous positions in wild-type HGF isoform 3 (e.g., SEQ ID NO: 1) are mutated. In some embodiments, the HGF is a polypeptide comprising a mutation in an amino acid sequence as shown below in Table 1 and Table 2. Any combination of mutations shown in Table 1 and Table 2 can be included in the HGF polypeptides disclosed herein. In some embodiments, the HGF is a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 7-27, with or without the signal sequence (e.g., the amino acids corresponding to amino acids 1–31 of SEQ ID NOs: 7–27). In some embodiments, the HGF is a polypeptide consisting of an amino acid sequence selected fromAttorney Docket No.: 49082-0007WO1 the group consisting of SEQ ID NOs: 7-27, with or without the signal sequence (e.g., the amino acids corresponding to amino acids 1–31 of SEQ ID NOs: 7–27). The HGF polypeptides disclosed herein can be monomers or dimers. In some embodiments, the HGF is a covalent dimer of a polypeptide, or a polypeptide variant thereof, of any one of any one of SEQ ID NOs: 1-27, with or without the signal sequence (e.g., the amino acids corresponding to amino acids 1–31 of SEQ ID NOs: 1–27). Covalent dimers of the HGF polypeptides can be obtained by expressing in a suitable expression system (for example, yeast or E. coli) any one of the HGF polypeptide sequences of SEQ ID NOs: 1-27 (with or without the signal sequence (e.g., the amino acids corresponding to amino acids 1– 31 of SEQ ID NOs: 1–27)), or a variant thereof, where a single amino acid residues, in particular, the N-terminus amino acid residue, is replaced with a cysteine residue. The expressed and, e.g., signal sequence cleaved, HGF polypeptide monomer can be induced to dimerize via the formation of a disulfide bond between the introduced cysteine residue (see, for example, Liu, et al., 2014, FEBS Letters, 588:4831-4837; Jones II, et al., 2011, Proc. Natl. Acad. Sci., 108:13035-13040; and USSN 15 / 365,514). In some cases, the HGF polypeptides described herein comprise one or more post- translational modifications, including for example, one or more of the following:Attorney Docket No.: 49082-0007WO1The HGF polypeptides disclosed herein can include a substitution with a naturally occurring amino acid or a non-naturally occurring amino acid including, but not limited to, hydroxyproline (Hyp), beta-alanine, citrulline (Cit), ornithine (Orn), norleucine (Nle), 3- nitrotyrosine, nitroarginine, and pyroglutamic acid (Pyr). In some cases, the HGF polypeptides are modified post-translationally, e.g., as shown in the table above. As used herein, the term “sequence identity” refers to the percentage of identical residues between two polypeptide or nucleic acid sequences. One skilled in the art can readily determine sequence identity to an amino acid of HGF or FGF, for example, by using the Basic Local Alignment Search Tool (BLAST) available online at https: / / blast.ncbi.nlm.nih.gov / Blast.cgi by inputting the amino acid (or nucleic acid) sequences of HGF or FGF and the polypeptide (or nucleic acid) in question. The program BLAST can, for example, be used to align two sequences (with default parameters) as described by Tatiana A. Tatusova and Thomas L. Madden (1999), "Blast 2 sequences - a new tool for comparing protein and nucleotide sequences", FEMS Microbiol Lett. 174:247-250. Various other algorithms and software that may be used to obtain alignments of amino acid or nucleotide sequences are well-known in the art. These include, but are not limited to, ALIGN, Megalign, BestFit, GCG Wisconsin Package, and variants thereof. In some embodiments, polypeptides of the invention have at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 97%, at least about 98%, or at least about 99% homology with HGF, as ascertained for example by using the program BLAST to align two sequences (default parameters). In some instances, the sequences are substantially identical over the full length of the sequences being compared, for example, (i) the coding region of a nucleotide sequence or (ii) an amino acid sequence. As used herein, “polypeptides” or “proteins” are polymers of amino acids having, for example, from 2 to about 1000 or more amino acid residues. In some embodiments, “polypeptides” have from 10 to about 130 amino acids, from 10 to about 220 amino acids, from 10 to about 500 amino acids, or from 10 to about 730 amino acids. Any naturally occurring or synthetic amino acid can form the polypeptide. Polypeptides can also include modifications such as glycosylations and other moieties. In some embodiments, polypeptides of the invention have the ability to selectively bind to target polypeptides based on, forAttorney Docket No.: 49082-0007WO1 example, amino acid sequence of the target, such as amino acid sequences of the N- or C- terminus.Attorney Docket No.: 49082-0007WO1li.w3 R G G 72 Ntm 3 1 D D Dso nrafir se 0 5 3 E K 2ITacn 1vFereff8 3 G Y H 2 DHi2 d 1nisyln A 2 5 4 5 1n 1 1 1 1 51 51 31 02 21 51 0 F VoO.A 1it eatpy.ut-dep b 51 12 61 81 91 02 61 82 41 71 89 Wmdlinecwiatnee si r mro 1 6 u 2si f1 1 1 1 1 1 1 1 1 1 9 Cqe : eo S O usI.di1 N seelDI rDID O 0 1 2 3 4 5 6Ib Q F Q 2 7 8 9 1 1 Q O 2 7 8 9 0a E G E N 1 1 1 1 1 E N 1 T S H S SAttorney Docket No.: 49082-0007WO1 D YW Wta9 ht7 G R n R R R R Rae8 K R R R E E R N V VV V yt-dli3 w 3 R D D D.stmo nrfT T T air s0 E aec3 v n FerAsGefnfiA 31 1 7 3 3 6 4 9 s oH dA 1 1 1 1 1 1 nitoitatnisyln atu noOum .edip 7 6 0 8 7 1 6 4 itatpy.b 1 1 2 1 1 2 1 1 R mricaaut-ddlen p omien iaRsiecwtabmeane si r mrof1 1 1 1 1 1 1 1 1 ffu 2 oor:sieo eqesS O uIreb b.N dise11 21 31 41 51 6 mmu21 uen nlDI rQ O 7 8 9 0 ::ba Q F G E SN2 1 1 1 2 12 22 32 42 p A EDb A T S HIAttorney Docket No.: 49082-0007WO1 R S A ER R 731 K R R R R R R R R R R R 531 S N N G G 231 K N N N N N N N R R N K 031IV V V V 61 51 2 7 1 21 N D D D D D D D D D D Dssno nitoitatu4 3 2 12 41 2at1 D A A ummd riicaapo n 2esi1 1 1 11 F Sabmaf foorereb O bm5 6 7 Q 7 8 9 0 1 2 3 4 5 6 7 muu 2 2 2 EN2 1 1 1 2 2 2 2 n SD2 2 2 2 I n: :pA b AAttorney Docket No.: 49082-0007WO1 COMPOSITIONS AND METHODS FOR PRODUCTION OF CELL LINES CO-EXPRESSING HEPSIN AND HGF Provided herein, among other things, are compositions and methods for the production of hepsin activated HGF, e.g., by way of cells engineered to co-express hepsin and pro-HGF. Nucleic Acid Expression Constructs Provided herein are nucleic acid expression construct(s) comprising nucleic acid sequences encoding hepsin (e.g., as described herein), and / or HGF (e.g., pro- HGF, e.g., as described herein), each with or without a signal sequence (endogenous or exogenous), e.g., for expression in a host cell. In some cases, the signal sequence is present at the 5’ end of the nucleic acid sequence (N-terminus of the encoded protein). In some cases, the signal sequence is present at the 3’ end of the nucleic acid sequence (C-terminus of the encoded protein). In some cases, the nucleic acid sequence encoding hepsin comprises a signal sequence. In some cases, the signal sequence is exogenous. In some cases, the nucleic acid sequence encoding HGF comprises a signal sequence. In some cases, the signal sequence is a native signal sequence (e.g., the native HGF signal peptide corresponding to amino acids 1–31 of, e.g., SEQ ID NO:1). In some cases, the signal sequence is not the native signal sequence. In some cases, the signal sequence is selected from the following:Attorney Docket No.: 49082-0007WO1In some embodiments, the nucleic acid sequence encoding hepsin and the nucleic acid sequence encoding HGF are on the same construct. In some cases, the nucleic acid sequence encoding hepsin and the nucleic acid sequence encoding HGF are on different constructs. In some cases, the nucleic acid expression construct comprises a promoter operably linked to the nucleic acid sequence encoding hepsin and / or HGF. In some cases, the promoter is selected from the group consisting of CMV (human or simian), EF1a, SV40, PGK1 (human or mouse), Ubc, Human beta actin, CAG, TRE, UAS, CaMKIIa, H1, RSV, UB, and U6. In some cases, the promoter is a cytomegalovirus immediate-early enhancer / promoter (human or simian). In some cases, the nucleic acid construct(s) are introduced into the host cell lines containing multiple docking sites, e.g., as described in WO2021247671 and / or below. Thus, in some cases, the nucleic acid construct(s) comprise nucleic acid sequence(s) (which may be termed “expression construct insertion elements”) that are compatible with the dock site insertion elements as described in WO2021247671 or below. In some cases, e.g., where the dock site does not comprise a promoter, the nucleic acid expression construct(s) comprise the following elements in operable association: first promoter sequence - selectable marker sequence - second promoter sequence - nucleic acid sequence encoding hepsin and / or HGF - poly A signal sequence. In some cases, e.g., where the dock site does not comprise a promoter, the nucleic acid expression construct(s) comprise the following elements in operable association, in 5’ to 3’ order:Attorney Docket No.: 49082-0007WO1 first promoter sequence - selectable marker sequence - second promoter sequence - nucleic acid sequence encoding hepsin and / or HGF - poly A signal sequence. In some cases, e.g., where the dock site comprises an exogenous promoter, the nucleic acid expression construct(s) comprise the following elements in operable association: selectable marker sequence - second promoter sequence - nucleic acid sequence encoding hepsin and / or HGF - poly A signal sequence. In some cases, e.g., where the dock site comprises an exogenous promoter, the nucleic acid expression construct(s) comprise the following elements in operable association, from 5’ to 3’ order: selectable marker sequence - second promoter sequence - nucleic acid sequence encoding hepsin and / or HGF - poly A signal sequence. In some cases, the construct(s) do not comprise a poly A signal sequence between the selectable marker sequence and second promoter sequence. In some cases, the selectable marker is adjacent to the second promoter. In some cases, the second promoter is adjacent to the nucleic acid sequence encoding hepsin and / or HGF. In this context, the term “adjacent” means that there is no intervening functional element or intron between the listed components. In some cases, the nucleic acid expression construct(s) further comprises at least one expression construct insertion element at a position or positions selected from the group consisting of 5’ to the first promoter, 3’ to the poly A signal sequence, between the first promoter and the poly A signal sequence, between the selectable marker and the second promoter sequence, and both 5’ to the first promoter and 3’ to the poly A signal sequence. Suitable constructs include the following non-limiting examples: 1) expression construct insertion element - first promoter sequence (optional depending on whether the dock site already comprises an exogenous promoter sequence) - selectable marker sequence - second (i.e., internal) promoter sequence - nucleic acid sequence encoding hepsin and / or HGF - poly A signal sequence;Attorney Docket No.: 49082-0007WO1 2) first promoter sequence (optional depending on whether the dock site already comprises an exogenous promoter sequence) - selectable marker sequence - second promoter sequence - nucleic acid sequence encoding hepsin and / or HGF - poly A signal sequence - expression construct insertion element; 3) expression construct insertion element - first promoter sequence (optional depending on whether the dock site already comprises an exogenous promoter sequence) - selectable marker sequence - second promoter sequence - nucleic acid sequence encoding hepsin and / or HGF - poly A signal sequence - expression construct insertion element; 4) first promoter sequence (optional depending on whether the dock site already comprises an exogenous promoter sequence)- selectable marker sequence - expression construct insertion element - second promoter sequence - nucleic acid sequence encoding hepsin and / or HGF - poly A signal sequence. In some cases, the constructs may include nucleic acid sequences encoding both hepsin and HGF. Suitable constructs for expressing both hepsin and HGF are shown in the following non-limiting examples: 1) expression construct insertion element - first promoter sequence (optional depending on whether the dock site already comprises an exogenous promoter sequence) - selectable marker sequence - second (i.e., internal) promoter sequence - nucleic acid sequence encoding one of hepsin or HGF - WPRE (optional) - poly A signal sequence - third promoter sequence or IRES - nucleic acid sequence encoding the other of hepsin or HGF - WPRE (optional) - poly A signal sequence 2) first promoter sequence (optional depending on whether the dock site already comprises an exogenous promoter sequence) - selectable marker sequence - second promoter sequence - nucleic acid sequence encoding one of hepsin or HGF - WPRE (optional) - poly A signal sequence - third promoter sequence - intron (optional) - nucleic acid sequence encoding the other of hepsin or HGF - WPRE (optional) - poly A signal sequence - expression construct insertion element 3) expression construct insertion element - first promoter sequence (optional depending on whether the dock site already comprises an exogenous promoterAttorney Docket No.: 49082-0007WO1 sequence) - selectable marker sequence - second promoter sequence - nucleic acid sequence encoding hepsin or HGF - WPRE (optional) - poly A signal sequence - third promoter sequence - intron (optional) - nucleic acid sequence encoding the other of hepsin or HGF - WPRE (optional) - poly A signal sequence - expression construct insertion element. 4) first promoter sequence (optional depending on whether the dock site already comprises an exogenous promoter sequence) - selectable marker sequence - expression construct insertion element - second promoter sequence - nucleic acid sequence encoding hepsin or HGF - WPRE - poly A signal sequence - third promoter sequence or IRES - nucleic acid sequence encoding the other of hepsin or HGF - WPRE - poly A signal sequence 5) expression construct insertion element - first promoter sequence (optional depending on whether the dock site already comprises an exogenous promoter sequence) - selectable marker sequence - second promoter sequence - nucleic acid sequence encoding hepsin or HGF - WPRE (optional) - poly A signal sequence - third promoter sequence - nucleic acid sequence encoding the other of hepsin or HGF- WPRE (optional) - poly A signal sequence - expression construct insertion element. 6) expression construct insertion element - first promoter sequence (optional depending on whether the dock site already comprises an exogenous promoter sequence) - selectable marker sequence - second promoter sequence - nucleic acid sequence encoding hepsin or HGF - WPRE (optional) - poly A signal sequence - third promoter sequence - intron- nucleic acid sequence encoding the other of hepsin or HGF - WPRE (optional) - poly A signal sequence - expression construct insertion element. In some cases, a mixture of different constructs is utilized. In some cases, the mixture of different constructs may comprise constructs as described above and constructs starting with the internal or second promoter (i.e., starting after and not including the selectable marker). The nucleic acid construct(s) described herein may be utilized with many different vectors and vectors systems. These vectors and vectors system may preferably be used to introduce the nucleic acid expression constructs into the hostAttorney Docket No.: 49082-0007WO1 cells described below. Suitable vectors and vectors systems include, but are not limited to, viral gene insertion technologies such as retroviral, lentiviral and AAV systems as well as non-viral gene insertion technologies such as transposase, recombinase, integrase or CRISPR gene insertion. Specific examples of technologies / enzymes that can be used with nucleic acid construct(s) described herein include piggyback transposase systems, sleeping beauty transposase systems, Mosl transposase systems, Tol2 transposase systems, Leapin transposase systems, Lambda recombinase systems, FLP / FRT systems, Cre / Lox systems, MMLV integrase systems, Rep 78 integrase systems and CRISPR systems which can include nucleases or nickases as well as guide sequences. In some preferred embodiments, the system is a nucleic acid integration system with the proviso that the system is not a retroviral or lentiviral systems utilizing a retroviral or lentiviral LTR. In some cases, the expression construct insertion element comprises an attachment site (att). In some particular cases, the attachment site is attB. These attachment sites are utilized by the PhiC31 integrase, which is a recombinase enzyme and which can be provided in the host cell via a vector in preferred embodiments. These sites facilitate integration of the nucleic acid constructs into a dock site comprising attP attachment site. In other cases, attR and attL attachment sites may be utilized. In some cases, the expression construct insertion element comprises an Flp Recombination Target (FRT) site. These sites are utilized by the enzyme flippase, which is a recombinase enzyme and which can be provided in the host cell via a vector in preferred embodiments. These sites serve facilitate integration of nucleic acid constructs into dock sites comprising corresponding FRT sites. In some cases, the expression construct insertion element comprises a LoxP site. These sites are utilized by the Cre recombinase which can be provided in the host cell via a vector in preferred embodiments. These sites facilitate integration of nucleic acid constructs into dock sites comprising corresponding LoxP sites. In some cases, the expression construct insertion element is an HDR (homology directed repair) expression construct insertion element. HDR expression construct insertion elements are nucleic acid sequences that provide an area of homology (a “homology arm”) that base pair with corresponding homology arms in the dock site. These systems are preferably used with endonucleases that introduce double stranded breaks at a targeted site or sites, preferably flanked by the homologyAttorney Docket No.: 49082-0007WO1 arms. In some cases, the HDR expression construct insertion element comprises AAVS1 safe harbor locus homology arms. In some cases, the expression construct is specifically integrated in a dock site comprising the AAVSl safe harbor locus. The integration is facilitated by the Rep 78 endonuclease (nickase) which may be introduced into the host cell via a vector. The Rep 78 protein nickase promotes site- specific integration of nucleic acid sequences bearing homology arms corresponding to the AAVSl safe harbor locus. In some cases, the HDR expression construct insertion element comprises one or more homology arms that are exogenous sequences of from 30 to 1000 base pairs in length. These expression some cases embodiments, the nucleic acid construct is inserted at dock sites that comprise homology arms that are homologous to and base pair with the homology arms in the nucleic acid construct. For utilization with CRISPR gene editing systems, a CRISPR gene editing system-compatible nuclease is introduced into the host cell. The CRISPR gene editing system-compatible nuclease may be a wild-type endonuclease that creates a double-stranded break at a position determined by the guide RNA (and within the docking site) or a mutated nuclease (e.g., a nickase) that creates a single stranded break at a staggered positions within the dock site defined by two guide RNAs. In some cases, as described in PCT / US2019 / 064423, for example, the first promoter which drives selectable marker is a weak promoter. In some cases, a weak promoter is a promoter, preferably a constitutive promoter, that has activity that equal to or less than the activity of the SIN LTR promoter in a host of interest (e.g., a CHO cell) when operably linked to a selectable maker sequence. In some cases, a weak promoter is a promoter, preferably a constitutive promoter, that has activity that equal to or less than the activity of the human Ubiquitin C (UBC) promoter in a host of interest (e.g., a CHO cell) when operably linked to a selectable maker sequence. Suitable methods for assessing promoter strength are known in the art. See, e.g., Dandindorj et al. (2014) A Comparative Analysis of Constitutive Promoters Located in Adeno-Associated Viral Vectors, PLoS One 9(8): el06472; Zhang and Baum (2005) Evaluation of Viral and Mammalian Promoters for Use in Gene Delivery to Salivary Glands Mol. Ther. 12(3):528-536; Qin et al. (2010) Systematic Comparison of Constitutive Promoters and the Doxycycline-Inducible Promoter PLoS 5(5): el 0611; Jeyaseelan et al. (2001) Real-time detection of gene promoter activity:Attorney Docket No.: 49082-0007WO1 quantitation of toxin gene transcription, Nucleic Acids Research. 29 (12): 58e-58. In some cases, weak promoters have been altered to reduce promoter activity. Accordingly, in some cases, the vector(s) for expression of hepsin and / or HGF comprise a nucleic acid sequence encoding a selectable marker in operable association with a first weak promoter sequence or promoter sequence that has been altered to reduce promoter activity as compared to a non-altered or wild-type version of the first promoter sequence and a nucleic acid sequence encoding the hepsin and / or HGF operably linked to a second promoter sequence. The SIN LTR promoter sequence is one such example. Other promoter sequences described above may also be altered to reduce activity and provide a weak promoter or the weak promoter may be naturally occurring weak promoter such as the UBC promoter. In some cases, the nucleic acid constructs include a selectable marker. Suitable selectable markers include but are not limited to glutamine synthetase (GS), dihydrofolate reductase (DHFR) and the like. These genes are described in U.S. Pat. Nos. 5,770,359; 5,827,739; 4,399,216; 4,634,665; 5,149,636; and 6,455,275; all of which are incorporated herein by reference. In some cases, the selectable marker that is utilized is compatible with a host cell line that is deficient in the production of the enzyme encoded by the selectable marker nucleic acid sequence. Suitable host cell lines are described in more detail below. In other cases, the selectable marker is an antibiotic resistance marker, i.e., a gene that produces a protein that provides cells expressing this protein with resistance to an antibiotic. Suitable antibiotic resistance markers include genes that provide resistance to neomycin (neomycin resistance gene (neo)), hygromycin (hygromycin B phosphotransferase gene), puromycin (puromycin N-acetyl-transferase), and the like. In some cases, where secretion of the hepsin and / or HGF is desired, the nucleic acid constructs include a signal peptide sequence in operable association with the hepsin and / or HGF. The sequences of several suitable signal peptides are known to those in the art, including, but not limited to, those derived from tissue plasminogen activator, human growth hormone, lactoferrin, alpha-casein, and alpha-lactalbumin. In some cases, the nucleic acid constructs include an RNA export element (See, e.g., U.S. Pat. Nos. 5,914,267; 6,136,597; and 5,686,120; and WO99 / 14310, all of which are incorporated herein by reference) either 3' or 5' to the nucleic acid sequence encoding the hepsin and / or HGF.Attorney Docket No.: 49082-0007WO1 In some cases, the nucleic acid constructs include at least one internal ribosome entry site (IRES) sequence. The sequences of several suitable IRES's are available, including, but not limited to, those derived from foot and mouth disease virus (FDV), encephalomyocarditis virus, and poliovirus. The IRES sequence can be interposed between two transcriptional units (e.g., nucleic acids encoding different proteins or protein subunits) to form a polycistronic sequence so that the two transcriptional units are transcribed from the same promoter. In some cases, the nucleic acid constructs are incorporated into a nucleic acid expression vector. Vectors include, but are not limited to, nucleic acid molecules that are single-stranded, double-stranded, or partially double-stranded; nucleic acid molecules that comprise one or more free ends, no free ends (e.g. circular); nucleic acid molecules that comprise DNA, RNA, or both; and other varieties of polynucleotides known in the art. One type of vector is a “plasmid,” which refers to a circular double stranded DNA loop into which additional DNA segments can be inserted, such as by standard molecular cloning techniques. Another type of vector is a viral vector, wherein virally -derived DNA or RNA sequences are present in the vector for packaging into a virus (e.g. retroviruses, replication defective retroviruses, adenoviruses, replication defective adenoviruses, and adeno-associated viruses). Viral vectors also include polynucleotides carried by a virus for transfection into a host cell. 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) are 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.” Common expression vectors of utility in recombinant DNA techniques are often in the form of plasmids. Other suitable vectors include, but are not limited to, cosmids and Yeast Artificial Chromosomes. Accordingly, suitable nucleic acid expression vectors include, but are not limited to, transposon vectors as described above, as well as plasmid vectors, retroviral vectors, lentiviral vectors, AAV vectors, phage vectors, etc). It is contemplated that any vector may be used as long as it is replicable and viable in the host. In some cases, the vectors are mammalian expression vectors that compriseAttorney Docket No.: 49082-0007WO1 among other elements described herein an origin of replication, a suitable promoter and enhancer, and also any necessary ribosome binding sites, polyadenylation sites, splice donor and acceptor sites, transcriptional termination sequences, and 5' flanking non-transcribed sequences. Suitable plasmid vectors that may be adapted to incorporate the nucleic acid constructs described herein include specific plasmids systems for transposon vectors, FLP-FLT systems, Cre-lox systems, CRISPR-Cas9 systems, recombinase systems and integrase systems as well as plasmid vectors derived from pCIneo, pVAXl, pACT, Gateway plamids, pAdvantage, pBIND, pG51uc, pTNT, pTarget, pCat3, pSI, pCMV, pSV and the like. In some cases, the present invention provides host cells and host cell culture wherein the host cells express the hepsin and / or HGF from the nucleic acid constructs described above. Suitable cells include, for example, those described below. Cells The compositions and methods described herein are useful, for example, for host cell production of proteins, e.g., activated HGF, e.g., as described herein. Suitable host cell lines include, but are not limited to, Chinese hamster ovary cells (CHO-K1, ATCC CCl-61); bovine mammary epithelial cells (ATCC CRL 10274; bovine mammary epithelial cells); monkey kidney CV1 line transformed by SV40 (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture; see, e.g., Graham et ak, J. Gen Virol., 36:59

[1977] ); baby hamster kidney cells (BHK, ATCC CCL 10); mouse sertoli cells (TM4, Mather, Biol. Reprod. 23:243-251

[1980] ); monkey kidney cells (CV1 ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL- 1587); human cervical carcinoma cells (HELA, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); buffalo rat liver cells (BRL 3 A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells (Hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL51); TRI cells (Mather et al., Annals N.Y. Acad. Sci., 383:44-68

[1982] ); MRC 5 cells; FS4 cells; rat fibroblasts (208F cells); MDBK cells (bovine kidney cells); CAP (CEVEC's Amniocyte Production) cells; and a human hepatoma line (Hep G2). In some cases, the host cells are modified so that they are deficient, or are naturally deficient, in an enzyme activity that is required for growth or survival of theAttorney Docket No.: 49082-0007WO1 cells in the presence of a selection agent and which is provided by the selectable marker. For example, Chinese Hamster Ovary (CHO) cells have been modified to be deficient for GS. In some cases, e.g., where the vector includes a GS selectable marker, the host cell line is deficient in GS. In some cases, the GS deficient host cell line is the CHOZN® GS cell line available from Merck KGaA. In some cases, e.g., where the selectable marker is, for example, DHFR, the cell line is deficient for DHFR activity (i.e., DHFR). Suitable DHFR- cell lines include but are not limited to CHO-DG44 and derivatives thereof. In some cases, the cell(s) comprise multiple docks for gene insertion, e.g., as described in WO2021247671, which is hereby incorporated by reference in its entirety. Accordingly, in some cases, the host cell (or population of host cells) comprises from 1 to 1000, e.g., 1 to 500, 5 to 500, 5 to 250, 5 to 100, or 5 to 50 integrated docking sites, each docking site comprising at least one dock site insertion element. In some cases, the integrated docking sites are independently positioned throughout the genome. In some cases, the integrated docking sites are independent integrated docking sites that are separated from one another and positioned at independent sites within the genome. For example, the integrated docking sites may preferably be spread across a number of chromosome sin the genome. In other embodiments, the integrated docking sites may be present as concatemers which comprise multiple copies of the same DNA sequence linked in series. In some cases, the integrated docking sites comprise one or more insertion elements (which may be termed a “dock site insertion element”). In some cases, the dock site insertion elements are nucleic acid sequences that facilitate insertion of a nucleic acid sequence encoding a protein of interest at the dock site. Nucleic acid constructs that can be inserted into the dock sites in the host cells are described in detail above. In some cases, the insertion element is a recombinase dock site insertion element. Recombinase dock site insertion elements are nucleic acid sequences that are recognized and utilized by recombinase enzymes. For example, in some preferred embodiments, the recombinase dock site insertion element comprises an attachment site (att). In some particularly preferred embodiments, the attachment site is attP. These attachment sites are utilized by the PhiC31 integrase, which is a recombinase enzyme and which can be provided in the host cell via a vector in preferredAttorney Docket No.: 49082-0007WO1 embodiments. These dock sites serve as acceptors for integration of nucleic acid constructs comprising an attB attachment site. In other cases, attR and attL attachment sites are utilized In some cases, the recombinase dock site insertion element comprises an Flp Recombination Target (FRT) site. These sites are utilized by the enzyme flippase, which is a recombinase enzyme and which can be provided in the host cell via a vector in preferred embodiments. These dock sites serve as acceptors for integration of nucleic acid constructs comprising at the FRT site. In some cases, the recombinase dock site insertion element comprises a LoxP site. These sites are utilized by the Cre recombinase which can be provided in the host cell via a vector in preferred embodiments. These dock sites serve as acceptors for integration of nucleic acid constructs comprising the LoxP site. In some cases, the insertion element is an HDR (homology directed repair) dock site insertion element. HDR dock site insertion elements are nucleic acid sequences that provide an area of homology (a “homology arm”) that base pair with corresponding homology arms on the nucleic acid construct that is inserted at the site. These systems are preferably used with endonucleases that introduce double stranded breaks at a targeted site or sites, preferably flanked by the homology arms. In some cases, the HDR dock site insertion element is an AAVS1 safe harbor locus. In these embodiments, the dock site is used utilized by the Rep 78 endonuclease (nickase) which may be introduced into the host cell via a vector. The Rep 78 protein nickase promotes site-specific integration of nucleic acid sequences bearing homology arms corresponding to the AAVS1 safe harbor locus. In some cases, the HDR dock site insertion element comprises one or more homology arms that are exogenous sequences of from 30 to 1000 base pairs in length. These dock sites are preferably used in conjunction with CRISPR gene editing systems. In some embodiments, the dock site further comprises one or more sequences that are homologous to guide RNA sequences. In these embodiments, the nucleic acid construct that is inserted at the dock site preferably comprises homology arms that are homologous to and base pair with the homology arms in the dock site. For utilization with CRISPR gene editing systems, a CRISPR gene editing system- compatible nuclease is introduced into the host cell. The CRISPR gene editing system-compatible nuclease may be a wild-type endonuclease that creates a double- stranded break at a position determined by the guide RNA (and within the dockingAttorney Docket No.: 49082-0007WO1 site) or a mutated nuclease (e.g., a nickase) that creates a single stranded break at a staggered positions within the dock site defined by two guide RNAs. Suitable nucleases are described above in the discussion of nucleic acid expression constructs. In some cases, the docking site comprises a suitable promoter so that a promoter trap scheme is utilized when suitable nucleic acid constructs are introduced at the docking site. Suitable promoters include, but are not limited to, SIN-LTR, SV40, EFla, E. coli lac, E. coli trp, phage lambda PL, phage lambda PR, T3, T7, cytomegalovirus (CMV) immediate early, herpes simplex virus (HSV) thymidine kinase, alpha-lactalbumin, and mouse metallothionein-I promoter sequences. In some cases the promoter sequence is oriented at the dock site so that the promoter will drive expression from an inserted nucleic acid construct. In some cases, the promoter is oriented 5’ to the docking site. In some cases, the promoter is a SIN LTR. In these embodiments, the SIN-LTR and EPR are positioned 5’ to the dock site and a SIN LTR is positioned 3’ to the dock site. Any suitable host cell line, such as those described above, may comprise the docking sites. In some cases, integration at the dock sites requires an exogenous enzyme, e.g., expressed by the host cell. Suitable enzymes include, but are not limited to, recombinases (including integrases), endonucleases, and nickases. Accordingly, in some cases, host cells, e.g., as described herein comprise an exogenous nucleic acid sequence (or expression construct) for expression of a recombinase (including integrases), a endonuclease, and a nickase. In some cases, constructs for expressing the exogenous enzymes may be stably integrated into the genome of the host cell. In other cases, vectors for expressing the exogenous enzymes are transiently introduced into the host cell, for example with an extrachromosomal vector such as a plasmid. Suitable exogenous enzyme expression constructs and cells expressing the same are described, for example, in WO2021247671. Transduction / Transformation The nucleic acid construct(s) and vector(s) descried herein may be introduced into host cells described herein by any suitable means such as by transfection, transformation or transduction. In some cases, the nucleic acid construct(s) encoding hepsin and / or HGF are introduced into the host cell(s) at the same time. In some cases, the nucleic acidAttorney Docket No.: 49082-0007WO1 constructs encoding hepsin and / or HGF are introduced into the host cells in a serial manner (e.g., a nucleic acid construct encoding one of HGF or hepsin is introduced, a period of time is allowed to pass, and then a nucleic acid construct encoding the other of HGF or hepsin is introduced). In some cases, the cell(s) are transfected, transformed, or transduced at a ratio of from 1:1 to 1:1,000 (hepsin:HGF), e.g., 1:1 to 1:900, 1:1 to 1:800, 1:1 to 1:700, 1:1 to 1:600, 1:1 to 1:500, 1:1 to 1:400, 1:1 to 1:300, 1:1 to 1:200, 1:1 to 1:100, 1:1 to 1:90, 1:10 to 1:80, 1:1 to 1:70, 1:1 to 1:60, 1:1 to 1:50, 1:1 to 1:40, 1:1 to 1:30, 1:1 to 1:20, 1:1 to 1:10, 1:10 to 1:900, 1:10 to 1:800, 1:10 to 1:700, 1:10 to 1:600, 1:10 to 1:500, 1:10 to 1:400, 1:10 to 1:300, 1:10 to 1:200, 1:10 to 1:100, 1:10 to 1:90, 1:10 to 1:80, 1:10 to 1:70, 1:10 to 1:60, 1:10 to 1:50, 1:10 to 1:40, 1:10 to 1:30, 1:10 to 1:20, 1:20 to 1:1,000, 1:20 to 1:900, 1:20 to 1:800, 1:20 to 1:700, 1:20 to 1:600, 1:20 to 1:500, 1:20 to 1:400, 1:20 to 1:300, 1:20 to 1:200, 1:20 to 1:100, 1:20 to 1:90, 1:20 to 1:80, 1:20 to 1:70, 1:20 to 1:60, 1:20 to 1:50, 1:20 to 1:40, 1:20 to 1:30, 1:30 to 1:1,000: 1:30 to 1:900, 1:30 to 1:800, 1:30 to 1:700, 1:30 to 1:600, 1:30 to 1:500, 1:30 to 1:400, 1:30 to 1:300, 1:30 to 1:200, 1:30 to 1:100, 1:30 to 1:90, 1:30 to 1:80, 1:30 to 1:70, 1:30 to 1:60, 1:30 to 1:50, 1:30 to 1:40, 1:40 to 1:1,000: 1:40 to 1:900, 1:40 to 1:800, 1:40 to 1:700, 1:40 to 1:600, 1:40 to 1:500, 1:40 to 1:400, 1:40 to 1:300, 1:40 to 1:200, 1:40 to 1:100, 1:40 to 1:90, 1:40 to 1:80, 1:40 to 1:70, 1:40 to 1:60, 1:40 to 1:50, 1:50 to 1:1,000, 1:50 to 1:900, 1:50 to 1:800, 1:50 to 1:700, 1:50 to 1:600, 1:50 to 1:500, 1:50 to 1:400, 1:50 to 1:300, 1:50 to 1:200, 1:50 to 1:100, 1:50 to 1:90, 1:50 to 1:80, 1:50 to 1:70, 1:50 to 1:60, 1:60 to 1:1,000: 1:60 to 1:900, 1:60 to 1:800, 1:60 to 1:700, 1:60 to 1:600, 1:60 to 1:500, 1:60 to 1:400; 1:60 to 1:300; 1:60 to 1:200; 1:60 to 1:100; 1:60 to 1:90, 1:60 to 1:80, 1:60 to 1:70,1:70 to 1:1,000, 1:70 to 1:900, 1:70 to 1:800, 1:70 to 1:700, 1:70 to 1:600, 1:70 to 1:500, 1:70 to 1:400, 1:70 to 1:300, 1:70 to 1:200, 1:70 to 1:100, 1:70 to 1:90, 1:70 to 1:80, 1:80 to 1:1,000, 1:80 to 1:900, 1:90 to 1:800, 1:90 to 1:700, 1:90 to 1:600, 1:90 to 1:500, 1:80 to 1:400, 1:80 to 1:300, 1:80 to 1:200, 1:80 to 1:100, 1:80 to 1:90, 1:90 to 1:1,000, 1:90 to 1:900, 1:90 to 1:800, 1:90 to 1:700, 1:90 to 1:600, 1:90 to 1:500, 1:90 to 1:400, 1:90 to 1:300, 1:90 to 1:200, 1:90 to 1:100, 1:100 to 1:1,000, 1:100 to 1:900, 1:100 to 1:800, 1:100 to 1:700, 1:100 to 1:600, 1:100 to 1:500, 1:100 to 1:400, 1:100 to 1:300, 1:100 to 1:200, 1:200 to 1:1,000, 1:200 to 1:900, 1:200 to 1:800, 1:200 to 1:700, 1:200 to 1:600, 1:200 to 1:500, 1:200 to 1:400, 1:200 to 1:300, 1:300 to 1:1,000, 1:300 to 1:900, 1:300 to 1:800, 1:300 to 1:700, 1:300 to 1:600, 1:300 to 1:500, 1:300 to 1:400,Attorney Docket No.: 49082-0007WO1 1:300, 1:400 to 1:1,000, 1:400 to 1:900, 1:400 to 1:800, 1:400 to 1:700, 1:400 to 1:600, 1:400 to 1:5001:500 to 1:1,000, 1:500 to 1:900, 1:500 to 1:800, 1:500 to 1:700, 1:500 to 1:600, 1:600 to 1:1,000, 1:600 to 1:900, 1:600 to 1:800, 1:600 to 1:700, 1:700 to 1:1,000, 1:700 to 1:900, 1:700 to 1:800, 1:800 to 1:1,000, 1:800 to 1:900, or 1:900 to 1:1,000 hepsin:HGF. In some cases, after transfection or transduction, the cells are allowed to multiply, and are then trypsinized and re-plated. In some cases, individual colonies are then selected to provide clonally selected cell lines. In some cases, the clonally selected cell lines are screened by Southern blotting or PCR assays to verify that the desired number of integration events has occurred (e.g., in the ranges described above for transfection, transformation, or transduction). It is also contemplated that clonal selection allows the identification of superior protein producing cell lines. In some cases, the cells are not clonally selected following transfection. SEQUENCES, MUTANTS, AND VARIANTS Variants of the polypeptide and nucleic acid sequences e.g., of hepsin and / or HGF, are contemplated herein. In some cases, the variant nucleic acid sequence comprises one or more single nucleotide variant(s), insertion(s), deletion(s), indel(s), duplication(s), inversion(s), expansion(s), or any combination thereof, e.g., as compared to a reference nucleic acid sequence. In some cases, the amino acid sequence comprises one or more substitution mutation(s) (e.g., conservative substitution mutation(s), insertion(s), deletion(s), frameshift(s), or any combination thereof, e.g., in comparison to a reference amino acid sequence. In some cases, the polypeptide or nucleic acid sequences described herein have at least 80%, e.g., at least 85%, 90%, 95%, 98%, or 100% identity to a polypeptide or nucleic acid sequence provided herein, e.g., has differences at up to 1%, 2%, 5%, 10%, 15%, or 20% of the residues of the sequence provided herein replaced, e.g., with conservative mutations, e.g., including or in addition to the mutations described herein. In preferred embodiments, the variant retains desired activity of the parent, e.g., in the case of a hepsin mutant or variant, proteolytic activity, and in the case of HGF, binding to c-MET and / or activation of the MAPK pathway in epithelial cells (e.g., human corneal epithelial cells).Attorney Docket No.: 49082-0007WO1 In some cases, variant polypeptide sequence described herein comprise conservative substitutions, e.g., substitutions within the following groups: glycine, alanine; valine, isoleucine, leucine; aspartic acid, glutamic acid, asparagine, glutamine; serine, threonine; lysine, arginine; and phenylalanine, tyrosine. EXAMPLES The invention is further described in the following examples, which do not limit the scope of the invention described in the claims. Example 1: Plasmids and Transduction Methods used in the Examples Transductions were performed using either GPExTM(Bleck, “An Alternative Method for the Rapid Generation of Stable, High-Expressing Mammalian Cell Lines,” BioProcess J. 5(4):36–42 (2006)) to generate cell pools or GPExTMLighning. For GPExTM, briefly, replication-defective retroviral vectors, derived from Moloney murine leukemia virus (MLV) and pseudotyped with vesicular stomatitis virus G protein (VSV-G), are used to stably insert single copies of genes into dividing cells. Retrovectors deliver genes coded as RNA that, after entering the cell, are reverse transcribed to DNA and integrated stably into the genome of the host cell. Two enzymes, reverse transcriptase and integrase, provided transiently in the vector particle, perform this function. These integrated genes are maintained through subsequent cell divisions as if they were endogenous cellular genes. Regular GPExTMuses a derivative of the CHO-S cell line, whereas the GPExTMLightning process (see, e.g., WO2021247671, which is hereby incorporated by reference in its entirety) uses the CHOZn® GS knockout cell line as the starting base CHO line. GPExTMLightning also utilizes a novel gene insertion technology, allowing better control over the ratio of the HGF gene and the hepsin gene in the pooled cell lines. The GPExTMLightning technology uses a recombinase to specifically insert genes into “dock” sequences recognized by the recombinase and placed into the cell line using the GPEx process. Approximately 150-200 “dock” sequences are present in the cell line and available for gene insertion.Attorney Docket No.: 49082-0007WO1Example 2: Development of HGF producing CHO cells Transductions of CHO cells was performed in triplicate using GPExTM, as described in Example 1, using three cycles of transduction for both the native signal peptide construct (pFCS-hHGF-WPRE-SIN) as well as the bovine alpha-lactalbumin signal peptide construct (pFCS-SPhHGF-WPRE-SIN). The pooled cell lines for each were expanded and analyzed in fed-batch culture conditions. Each of the pooled cell lines were compare for amount of gene copies using a gene copy index value that is a ratio of the number of copies of the transgene to a single copy endogenous CHO gene using quantitative real-time PCR. The gene copy index, generated by subtracting the transgene Ct from a control Ct, reflects the number of transgene inserts that are present in the genome of the cell line. In general, gene index values are exponential – increasing by +1 reflects approximately 2-fold increase of the target relative to the control.. The higher the value the more copies of the gene, but a value of 1 does not mean a single gene copy is present. The pooled cell line gene copy index after each cycle of transduction was as follows:Attorney Docket No.: 49082-0007WO1The results indicated very similar copy numbers for the cell pools after 3 transductions and they were compared for HGF production. Since the gene copy numbers were similar, fed-batch production was performed with each cell pool to examine the HGF production and processing, using the conditions and sampling methodology as follows: Fed Batch Culture Conditions (Base Medium: G12.1 + 6 mM L-glutamine + 4 g / L PS307)Fed Batch Culture Sampling MethodologyAs shown in FIGs. 11A–11D, each of the cultures showed a very similar growth behavior typical of that of GPExTMpooled cell lines.Attorney Docket No.: 49082-0007WO1 PAGE analysis of expressed HGF was used to measure reduced and non- reduced forms of HGF in comparison to a control HGF standard (serum activated). As shown in FIG. 12, the non-reduced SDS-PAGE analysis of the productivity samples on day 4, 8 and end of culture showed that both the pooled cell lines appear to be producing HGF at a similar size to the HGF standard. However, there is much greater production from the cell pool expressing HGF with the native signal peptide. As shown in FIG. 13, the reduced SDS-PAGE analysis of the productivity samples on day 4,8 and end of culture showed that the majority of the native signal peptide sample is not fully cleaved to make fully processed HGF. The non-cleaved product is boxed. Example 3: Transduction of HGF producing CHO cells with hepsin The CHO cells transduced with the pFCS-hHGF-WPRE-SIN plasmid (Example 2) were further transduced with pFCS-newhHepsin-WPRE-SIN in triplicate using GPExTM, as described in Example 1. Cell pools were expanded and frozen after each transduction for further analysis. The pooled cell line gene copy index after each cycle of transduction was as follows:Surprisingly, very few copies of the hepsin gene were detected in the cell pools. Typically, one would see much higher gene copy index values with transductions. One would expect to see numbers similar to what was observed for HGF after each cycle of transduction. The numbers got higher with each cycle, but are significantly lower than the normal values observed with HGF. This can occur when the protein product is toxic to cells or inhibits cell growth when expressed at high levels. The cell pools were expanded and run in fed-batch productivity studies to examine HGF production and determine if any improvements were observed in regards to the production of fully processed HGF. As shown in FIG. 14A and FIG. 14B, no significant improvement in the amount of processed HGF was observed in these pooled cell lines. Non-reduced gels and reduced gels comparing cell poolAttorney Docket No.: 49082-0007WO1 production from the pool without hepsin, the 1 cycle hepsin transduced pool and the 2 cycle hepsin transduced pool, showed no significant difference in the amount of processed HGF. The red box highlights the un-processed HGF molecule in each of the 3 cell pool samples. Example 3: Co-transductions of GS knockout CHO cells with HGF and hepsin Four different pooled cell lines were produced using the GPExTMLightning process, as described in Example 1, as follows:As part of the GPExTMLightning process, after transfection of the integrase and the transgene constructs, selection by removal of glutamine from the media occurs to allow only cell lines containing the transgene (and the glutamine synthase) to survive. How fast cell lines recover from the selection can typically indicate how well the process worked as well as if there are any issues caused by transgene expression. Lines 1 and 4 responded “normally” and appeared to be healthy cultures. Lines 2 and 3 showed much slower recovery profiles and needed additional supplementation and attention in order to maintain growth. As shown in FIG. 15, the pooled cell line produced using a 100:1 ratio of HGF:Hepsin recovered from selection much faster than the other two cell pools and the recovery was highly correlated to the amount of hepsin the cells were in theory producing, consistent with the data that was observed using the GPEx process. The pools with poor recovery continued to have some growth issues. HGF production was analyzed for Lines 1 and 4. As was previously observed with the GPExTMcell lines only containing HGF, and shown in FIG. 16A and FIG. 16B, both Lines 1 and 4 showed good levels of HGF expression (FIG. 16A). Line 1 showed good HGF production, but very little processing / cleavage, whereas Line 4Attorney Docket No.: 49082-0007WO1 showed good HGF production and all the HGF appears to be processed / cleaved and looks very similar to the HGF standard produced using serum on SDS-PAGE gels (FIG. 16B). The expression of HGF was good in expansion media obtained from each of the two cell pools examined on the non-reduced gel. As observed with the original GPExTMexperiments that were performed in the CHO-S base cell line and now the GPExTMLightning data from the CHOZN GS knock-out cell line, very little processed HGF was observed in cell lines lacking hepsin. However, unlike the previous GPExTMexperiments that used hepsin, the 100:1 HGF:Hepsin cell pool now showed fully process HGF on the reduced SDS-PAGE gel. The 100:1 cell pool was showing normal cell growth and behavior and was fully processing the HGF. A fed-batch productivity study was carried out in 500 ml shake flasks for the two pooled cell lines (Line 1 and Line 4) with the following culture and sampling conditions: Fed Batch Culture Conditions (Base Medium: ExCell Advance Fed-Barch Media)Fed Batch Culture Sampling MethodologyAttorney Docket No.: 49082-0007WO1As shown in FIG. 17A and FIG. 17B, both cell pools showed very similar cell growth characteristics through the culture suggesting no issues with hepsin effecting the growth profile in the culture. HGF production was measured for the fed-batch cultures. As shown in FIGS. 18A–18D, the results were similar to those observed in the expansion media analysis, with little fully processed HGF in the HGF only pool and fully processed HGF in the 100:1 pool, except with respect to the presence of apparent extra cleavage in the 100:1 pool. However, the additional cleavage doesn’t seem to affect the behavior of the molecule in the non-reduced gel. Additional cleavage of HGF during the production / processing is selected against during clonal cell line selection process and the development of the cell culture and protein purification conditions. The material from the cell pools was purified using ion exchange chromatography. As shown in FIG. 19, purified products from each of the two cultures showed similar results to the non-purified culture media samples. HGF expression was high for both (left panel, non-reduced) and the HGF expressed without hepsin showed a low percentage of processed HGF, while the 100:1 pooled material showed full processing and the additional cleavage (right panel, reduced). Example 4: Peptide Mapping Peptide mapping was carried out on the HGF protein produced by Line 4. Samples were reduced and alkylated and then digested with each of Trypsin, Asp-N, and Glu-C. Searches were predicted digested species were carried out for each enzyme for both the un-cleaved HGF sequence and the hepsin-activated HGF sequence (cleaved into alpha and beta chains). The overall 3 enzyme sequence coverage was 99.7% (690 / 692 amino acids—99% coverage for Trypsin, 91% for Asp- N, and 85% for Glu-C). The tryspin digestion was not suitable for confirmation of hepsin cleavage. The lack of an Asp-N digest peptide corresponding to AA 455–503 of SEQ ID NO:32, suggests correct hepsin cleavage at the hepsin cleavage siteAttorney Docket No.: 49082-0007WO1 between R458 and V459 of SEQ ID NO:32. The lack of a Gly-C digest peptide corresponding to AA 435–490 of SEQ ID NO:32 also suggests correct hepsin cleavage at the hepsin cleavage site between R458 and V459 of SEQ ID NO:32. The HGF sequence of SEQ ID NO:32 contains four RV dipeptides—at the cleavage site between the alpha and beta chains (458–459), one within the alpha chain (337–338, “c1”), and two within the beta chain (594–595, “c2”; and 672–673, “c3”) and. Potential undesirable hepsin cleavage was analyzed by calculating the percent Asp-N and Glu-C cleaved at each of the four dinucleotide sites as follows: 100The calculated percent cleaved at each location was as follows:Example 5: Clonal Selection An initial round of clonal selection was carried out on the 100:1 HGF:Hepsin pool. For 76 of the clones, the number of gene copies of HGF was estimated using two different measures (Gene Copy Index and HGF Gene Copy Index in the table below). The estimated HGF copy number is 2^(the average of these two measures)., The presence of a hepsin gene (positive or negative) was also measured:Attorney Docket No.: 49082-0007WO1Attorney Docket No.: 49082-0007WO112 clones with 0, 1, or 2 copies of the Hepsin gene were selected for further analysis. The Estimated HGF Copy Number, Hepsin Gene Copy Number, and HGF:Hepsin Copy Number ratio for those clones are as follows:Attorney Docket No.: 49082-0007WO1All the identified clones with 2 copies of the Hepsin gene consistently overprocessed the HGF product in a miniature bioreactor fed-batch culture, resulting in unwanted cleavage products. All the clones with no exogenous Hepsin showed significant under-processing of the HGF. Clone #526 tended to be the clone that processed the HGF consistently to the most desirable form. For each of the three levels of hepsin copies (0, 1, and 2), the minimum, maximum, and mean Estimated HGF Copy Number of the corresponding clones as well as the minimum, maximum, and mean HGF:Hepsin Ratio are shown in the table below.Attorney Docket No.: 49082-0007WO1 SEQUENCES SEQ ID NO:1. Amino acid sequence of Homo sapiens HGF Isoform 3; (Identifier: P14210-3, Accession number NP_001010932.1) (dHGF) MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPAL KIKTKKVNTADQCANRCTRNKGLPFTCKAFVFDKARKQCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRNCIIGKGRSYKGTVSITKSGIKCQPWSSMIPHEHSYRGKDLQENY CRNPRGEEGGPWCFTSNPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTESGKICQ RWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAIKTCAD NTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPENFKCK DLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNYMGNLS QTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGNPLIPW DYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRYRNKHI CGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQLVYGPE GSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDGLLRVA HLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKMRMVLG VIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO:2. Amino acid sequence of Homo sapiens HGF Isoform 1; (Identifier: P14210-1, Accession number: NP_000592.3) MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPAL KIKTKKVNTADQCANRCTRNKGLPFTCKAFVFDKARKQCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRNCIIGKGRSYKGTVSITKSGIKCQPWSSMIPHEHSFLPSSYRGKD LQENYCRNPRGEEGGPWCFTSNPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO:3. Amino acid sequence of Homo sapiens HGF Isoform 2 (Identifier: P14210-2, Accession Number: NP_001010931.1): MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPAL KIKTKKVNTADQCANRCTRNKGLPFTCKAFVFDKARKQCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRNCIIGKGRSYKGTVSITKSGIKCQPWSSMIPHEHSFLPSSYRGKD LQENYCRNPRGEEGGPWCFTSNPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCET SEQ ID NO:4. Amino acid sequence of Homo sapiens HGF Isoform 4 (Identifier: P14210-4): MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPAL KIKTKKVNTADQCANRCTRNKGLPFTCKAFVFDKARKQCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRNCIIGKGRSYKGTVSITKSGIKCQPWSSMIPHEHSFLPSSYRGKDAttorney Docket No.: 49082-0007WO1 LQENYCRNPRGEEGGPWCFTSNPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KNMRDITWALN SEQ ID NO:5. Amino acid sequence of Homo sapiens HGF Isoform 5 (Identifier: P14210-5, NP_001010933.1): MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPAL KIKTKKVNTADQCANRCTRNKGLPFTCKAFVFDKARKQCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRNCIIGKGRSYKGTVSITKSGIKCQPWSSMIPHEHSYRGKDLQENY CRNPRGEEGGPWCFTSNPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTESGKICQ RWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAIKTCET SEQ ID NO:6. Amino acid sequence of Homo sapiens HGF Isoform 6 (Identifier: P14210-6, NP_001010934.1): MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPAL KIKTKKVNTADQCANRCTRNKGLPFTCKAFVFDKARKQCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRNCIIGKGRSYKGTVSITKSGIKCQPWSSMIPHEHSFLPSSYRGKD LQENYCRNPRGEEGGPWCFTSNPEVRYEVCDIPQCSEGK SEQ ID NO: 7. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPAL EIKTKKANTADQCANRCTRSKGLPFTCKAFVFDKARKRCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRDCIIGKGRSYRGTVSITKSGIKCQPWSAMIPHEHSFLPSSYRGED LRENYCRNPRGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO: 8. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPAL KIKTEKANTADQCANRCIRNKGLPFTCKAFVFDKARKRCLWFPVNSMSSGVKKEFGH EFDLYENKDYTRNCIVGNGRSYRGTVSTTKSGIKCQPWSAMIPHEHSFLPSSYRGED LRENYCRNPRGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQSAttorney Docket No.: 49082-0007WO1 SEQ ID NO: 9. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPAL KIKTEKANTADQCANRCTRSKGLPFTCKAFVFDKARKRCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRDCIVGNGRSYRGTVSITKSGIECQPWSAMIPHEHSFLPSSYRGED LRENYCRNPRGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO: 10. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAKGQGKRRNTIHEFKKSAKTTLIKIDPAL KIKTEKADTADQCANRCTRSKGLPFTCKAFVFDKARKQCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRDCIVGNGRSYRGTVSVTKSGIKCQPWSSMIPHEHSFLPSSYRGED LRENYCRNPRGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO: 11. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNAIHEFKKSAKATLIKIDPAL KIKTEKANTADQCANRCTRSKGLPFTCKAFVFDKARKRCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRDCIVGNGRSYRGTVSITKSGIECQPWSSMIPHEHSFLPSSYRGED LQENYCRNPWGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO: 12. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPAL KIKTEKANTADQCANRCTRSKGLPFTCKAFVFDKARKRCLWFPFNSMSSGVKKEFGH EFDLYENKDYTRNCIVGNGRSYRGTVSITKSGIECQPWSAMIPHEHSFLPSSYQGED LRENYCRNPRGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAIAttorney Docket No.: 49082-0007WO1 KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO: 13. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPAL KIKTKKVNTADQCANRCTRNKGLPFTCKAFVFDKARKQCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRNCIIGKGRSYKGTVSITKSGIKCQPWSSMIPHEHSFLPSSYRGKD LQENYCRNPRGEEGGPWCFTSNPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO: 14. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRDAIHECKRSAKTTLIKIDPAL KIKTEKANTADQCANRCTRNKGLPSTCKAFVFDKARKRRLRFPFNSMSSGVKKEFGH EFDLYENKDYTRNCIVGKGRSYRGTVSTTKSGIKCQPWSAMIPHEHSFLPSSYRGED LRENYCRNPRGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO: 15. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQGKRRNTIHEFKKSAKTTLIKIDPAL KIKTEKVNTADQCANRCTRSKGLPFTCKAFVFDKARKRCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRDCIIGRGRSYRGTVSITKSGIKCQPWSAMIPHEHSFLPSSYRGED LRENYCRNPRGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDGAttorney Docket No.: 49082-0007WO1 LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO: 16. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPHAEGQRKRRNTIHEFKKSAKTTLIKIDPAL KIKTEKANTADQCANRCTRSKGLPFTCKAFVFDKARKRCLWFPFNSMSSGVKKEFGH EFDLYENKDYTRNCIVGNGRSYRGTVSTTKSGIKCQPWSAMIPHEHSFLPSSYRGED LRENYCRNPRGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO: 17. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPAL KIKTEKANTADQCANRCTRSRGLPFTCKAFVFDKARKQCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRDCIIGNGRSYRGTVSVTKSGIKCQPWSAMIPHEHSFLPSSYRGED LRENYCRNPRGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO: 18. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPAL KIKTEKANTADQCANRCTRNKGLPFTCKAFVFDKARKRCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRDCIVGNGRSYRGTVSITKSGIKCQPWSSMIPHEHSFLPSSYRGED LRENYCRNPRGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO: 19. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSVKTTLIKIDPAL KIKTEKANTADQCANRCTRSKGLPFTCKAFVFDKARKRCLWFPVNSMSSGVKKESGHAttorney Docket No.: 49082-0007WO1 EFDLYENKDYIRDCIVGNGRSYRGTVSTTKSGIKCQPWSAMIPHEHSFLPSSYRGED LRENYCRNPRGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO: 20. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPAL KIKTEKANTADQCANRCTRSKGLPFTCKAFVFDKARKRCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRDCIVGNGRSYRGTVSITKSGIKCQPWSSMIPHEHSFLPSSYRGED LRENYCRNPWGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO: 21. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPAL RIKTEKANTADQCANRCTRSRGLPFTCKAFVFDKARKRCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRDCIIGNGRSYRGTVSITKSGIKCQPWSSMIPHEHSFLPSSYRGED LRENYCRNPRGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO: 22. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPAL KIKTEKANTADQCANRCTRSRRLPFTCKAFVFDKARKRCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRDCIIGKGRSYRGTVSVTKSGIECQPWSAMIPHEHSFLPSNYRGED LRENYCRNPRGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGNAttorney Docket No.: 49082-0007WO1 PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO: 23. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPAL KIKTEKANTADQCANRCTRSKGLPFTCKAFVFDKARKRCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRDCIVGNGRSYRGTVSITKSGIECQPWSAMIPHEHSFLPSSYRGED LRENYCRNPRGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO: 24. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQRKRRNTIHEFKKSAKTTLIKIDPAL KIKTKKVDTADQCANRCTRNKGLPFTCKAFVFDKARKRCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRDCIIGNGRSYRGTVSITKSGIKCQPWSSMIPHEHSFLPSSYRGED LRENYCRNPRGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO: 25. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQGKRRNTIHEFKKSAKTTLIKIDPAL RIKTEKANTADQCANRCTRSKGLPFTCKAFVFDKARKRCLWFPFNSMSSGVKKEFGH EFDLYENKAYIRDCIIGRGRNYRGTVSITKSGIKCQPWSAMIPHEHSFLPSSYRGED LRENYCRNPRGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQSAttorney Docket No.: 49082-0007WO1 SEQ ID NO: 26. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAKGQRKRRNTIHEFKKSAKTTLIKIDPAL EIKTEKVNTADQCANRCIRNKGLPFTCKAFVFDKARKRCLWFPFNSMSSGVKKEFGH EFDLYENKAYIRDCIIGRGRNYRGTVSITKSGIKCQPWSAMIPHEHSFLPSSYRGED LRENYCRNPRGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO: 27. HGF Variant: MWVTKLLPALLLQHVLLHLLLLPIAIPYAEGQGKRRNTIHEFKKSAKTTLIKIDPAL KIKTEKVNTADQCANRCTRSKGLPFTCKAFVFDKARKRCLWFPFNSMSSGVKKEFGH EFDLYENKDYIRDCIIGNGRSYRGTVSITKSGIKCQPWSAMIPHEHSFLPSSYRGED LRENYCRNPRGEEGGPWCYTSDPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTES GKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAI KTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPE NFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNY MGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGN PLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRY RNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQL VYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDG LLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKM RMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILTYKVPQS SEQ ID NO:28 (>sp|P05981|HEPS_HUMAN Serine protease hepsin OS=Homo sapiens OX=9606 GN=HPN PE=1 SV=1) MAQKEGGRTVPCCSRPKVAALTAGTLLLLTAIGAASWAIVAVLLRSDQEPLYPVQVS SADARLMVFDKTEGTWRLLCSSRSNARVAGLSCEEMGFLRALTHSELDVRTAGANGT SGFFCVDEGRLPHTQRLLEVISVCDCPRGRFLAAICQDCGRRKLPVDRIVGGRDTSL GRWPWQVSLRYDGAHLCGGSLLSGDWVLTAAHCFPERNRVLSRWRVFAGAVAQASPH GLQLGVQAVVYHGGYLPFRDPNSEENSNDIALVHLSSPLPLTEYIQPVCLPAAGQAL VDGKICTVTGWGNTQYYGQQAGVLQEARVPIISNDVCNGADFYGNQIKPKMFCAGYP EGGIDACQGDSGGPFVCEDSISRTPRWRLCGIVSWGTGCALAQKPGVYTKVSDFREW IFQAIKTHSEASGMVTQL SEQ ID NO:29 Extracellular domain of P05981 RSDQEPLYPVQVSSADARLMVFDKTEGTWRLLCSSRSNARVAGLSCEEMGFLRALTH SELDVRTAGANGTSGFFCVDEGRLPHTQRLLEVISVCDCPRGRFLAAICQDCGRRKL PVDRIVGGRDTSLGRWPWQVSLRYDGAHLCGGSLLSGDWVLTAAHCFPERNRVLSRW RVFAGAVAQASPHGLQLGVQAVVYHGGYLPFRDPNSEENSNDIALVHLSSPLPLTEY IQPVCLPAAGQALVDGKICTVTGWGNTQYYGQQAGVLQEARVPIISNDVCNGADFYG NQIKPKMFCAGYPEGGIDACQGDSGGPFVCEDSISRTPRWRLCGIVSWGTGCALAQK PGVYTKVSDFREWIFQAIKTHSEASGMVTQLAttorney Docket No.: 49082-0007WO1 SEQ ID NO:30 Serine protease hepsin catalytic chain of P05981 IVGGRDTSLGRWPWQVSLRYDGAHLCGGSLLSGDWVLTAAHCFPERNRVLSRWRVFA GAVAQASPHGLQLGVQAVVYHGGYLPFRDPNSEENSNDIALVHLSSPLPLTEYIQPV CLPAAGQALVDGKICTVTGWGNTQYYGQQAGVLQEARVPIISNDVCNGADFYGNQIK PKMFCAGYPEGGIDACQGDSGGPFVCEDSISRTPRWRLCGIVSWGTGCALAQKPGVY TKVSDFREWIFQAIKTHSEASGMVTQL SEQ ID NO:31 Signal Peptide of SEQ ID NO:1 MWVTKLLPALLLQHVLLHLLLLPIAIPYAEG SEQ ID NO:32 Un-cleaved Alpha and Beta chain peptide of SEQ ID NO:1 QRKRRNTIHEFKKSAKTTLIKIDPALKIKTKKVNTADQCANRCTRNKGLPFTCKAFV FDKARKQCLWFPFNSMSSGVKKEFGHEFDLYENKDYIRNCIIGKGRSYKGTVSITKS GIKCQPWSSMIPHEHSYRGKDLQENYCRNPRGEEGGPWCFTSNPEVRYEVCDIPQCS EVECMTCNGESYRGLMDHTESGKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPD GQPRPWCYTLDPHTRWEYCAIKTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIW NGIPCQRWDSQYPHEHDMTPENFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCS QIPNCDMSHGQDCYRGNGKNYMGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKL NENYCRNPDDDAHGPWCYTGNPLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQ LRVVNGIPTRTNIGWMVSLRYRNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLG IHDVHGRGDEKCKQVLNVSQLVYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTI PEKTSCSVYGWGYTGLINYDGLLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEK IGSGPCEGDYGGPLVCEQHKMRMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKII LTYKVPQS SEQ ID NO:33 Alpha chain peptide of SEQ ID NO:1 QRKRRNTIHEFKKSAKTTLIKIDPALKIKTKKVNTADQCANRCTRNKGLPFTCKAFV FDKARKQCLWFPFNSMSSGVKKEFGHEFDLYENKDYIRNCIIGKGRSYKGTVSITKS GIKCQPWSSMIPHEHSYRGKDLQENYCRNPRGEEGGPWCFTSNPEVRYEVCDIPQCS EVECMTCNGESYRGLMDHTESGKICQRWDHQTPHRHKFLPERYPDKGFDDNYCRNPD GQPRPWCYTLDPHTRWEYCAIKTCADNTMNDTDVPLETTECIQGQGEGYRGTVNTIW NGIPCQRWDSQYPHEHDMTPENFKCKDLRENYCRNPDGSESPWCFTTDPNIRVGYCS QIPNCDMSHGQDCYRGNGKNYMGNLSQTRSGLTCSMWDKNMEDLHRHIFWEPDASKL NENYCRNPDDDAHGPWCYTGNPLIPWDYCPISRCEGDTTPTIVNLDHPVISCAKTKQ LR SEQ ID NO:34 Beta chain peptide of SEQ ID NO:1 VVNGIPTRTNIGWMVSLRYRNKHICGGSLIKESWVLTARQCFPSRDLKDYEAWLGIH DVHGRGDEKCKQVLNVSQLVYGPEGSDLVLMKLARPAVLDDFVSTIDLPNYGCTIPE KTSCSVYGWGYTGLINYDGLLRVAHLYIMGNEKCSQHHRGKVTLNESEICAGAEKIG SGPCEGDYGGPLVCEQHKMRMVLGVIVPGRGCAIPNRPGIFVRVAYYAKWIHKIILT YKVPQS SEQ ID NO:35 nucleic acid sequence of HGF CDS in vector pFCS-hHGF-WPRE-SIN (new ori) 1008-146, CDI10.0002 ATGTGGGTGACCAAACTCCTGCCAGCCCTGCTGCTGCAGCATGTCCTCCTGCATCTC CTCCTGCTCCCCATCGCCATCCCCTATGCAGAGGGACAAAGGAAAAGAAGAAATACA ATTCATGAATTCAAAAAATCAGCAAAGACTACCCTAATCAAAATAGATCCAGCACTG AAGATAAAAACCAAAAAAGTGAATACTGCAGACCAATGTGCTAATAGATGTACTAGGAttorney Docket No.: 49082-0007WO1 AATAAAGGACTTCCATTCACTTGCAAGGCTTTTGTTTTTGATAAAGCAAGAAAACAA TGCCTCTGGTTCCCCTTCAATAGCATGTCAAGTGGAGTGAAAAAAGAATTTGGCCAT GAATTTGACCTCTATGAAAACAAAGACTACATTAGAAACTGCATCATTGGTAAAGGA CGCAGCTACAAGGGAACAGTATCTATCACTAAGAGTGGCATCAAATGTCAGCCCTGG AGTTCCATGATACCACACGAACACAGCTATCGGGGTAAAGACCTACAGGAAAACTAC TGTCGAAATCCTCGAGGGGAAGAAGGGGGACCCTGGTGTTTCACAAGCAATCCAGAG GTACGCTACGAAGTCTGTGACATTCCTCAGTGTTCAGAAGTTGAATGCATGACCTGC AATGGGGAGAGTTATCGAGGTCTCATGGATCATACAGAATCAGGCAAGATTTGTCAG CGCTGGGATCATCAGACACCACACCGGCACAAATTCTTGCCTGAAAGATATCCCGAC AAGGGCTTTGATGATAATTATTGCCGCAATCCCGATGGCCAGCCGAGGCCATGGTGC TATACTCTTGACCCTCACACCCGCTGGGAGTACTGTGCAATTAAAACATGCGCTGAC AATACTATGAATGACACTGATGTTCCTTTGGAAACAACTGAATGCATCCAAGGTCAA GGAGAAGGCTACAGGGGCACTGTCAATACCATTTGGAATGGAATTCCATGTCAGCGT TGGGATTCTCAGTATCCTCACGAGCATGACATGACTCCTGAAAATTTCAAGTGCAAG GACCTACGAGAAAATTACTGCCGAAATCCAGATGGGTCTGAATCACCCTGGTGTTTT ACCACTGATCCAAACATCCGAGTTGGCTACTGCTCCCAAATTCCAAACTGTGATATG TCACATGGACAAGATTGTTATCGTGGGAATGGCAAAAATTATATGGGCAACTTATCC CAAACAAGATCTGGACTAACATGTTCAATGTGGGACAAGAACATGGAAGACTTACAT CGTCATATCTTCTGGGAACCAGATGCAAGTAAGCTGAATGAGAATTACTGCCGAAAT CCAGATGATGATGCTCATGGACCCTGGTGCTACACGGGAAATCCACTCATTCCTTGG GATTATTGCCCTATTTCTCGTTGTGAAGGTGATACCACACCTACAATAGTCAATTTA GACCATCCCGTAATATCTTGTGCCAAAACGAAACAATTGCGAGTTGTAAATGGGATT CCAACACGAACAAACATAGGATGGATGGTTAGTTTGAGATACAGAAATAAACATATC TGCGGAGGATCATTGATAAAGGAGAGTTGGGTTCTTACTGCACGACAGTGTTTCCCT TCTCGAGACTTGAAAGATTATGAAGCTTGGCTTGGAATTCATGATGTCCACGGAAGA GGAGATGAGAAATGCAAACAGGTTCTCAATGTTTCCCAGCTGGTATATGGCCCTGAA GGATCAGATCTGGTTTTAATGAAGCTTGCCAGGCCTGCTGTCCTGGATGATTTTGTT AGTACGATTGATTTACCTAATTATGGATGCACAATTCCTGAAAAGACCAGTTGCAGT GTTTATGGCTGGGGCTACACTGGATTGATCAACTATGATGGCCTATTACGAGTGGCA CATCTCTATATAATGGGAAATGAGAAATGCAGCCAGCATCATCGAGGGAAGGTGACT CTGAATGAGTCTGAAATATGTGCTGGGGCTGAAAAGATTGGATCAGGACCATGTGAG GGGGATTATGGTGGCCCACTTGTTTGTGAGCAACATAAAATGAGAATGGTTCTTGGT GTCATTGTTCCTGGTCGTGGATGTGCCATTCCAAATCGTCCTGGTATTTTTGTCCGA GTAGCATATTATGCAAAATGGATACACAAAATTATTTTAACATATAAGGTACCACAG TCATAG SEQ ID NO:36 Hepsin CDS in vector pFCS-newhHepsin-WPRE- SIN (new ori) 1008-146, CI01.0004 ATGATGTCCTTTGTCTCTCTGCTCCTGGTTGGCATCCTATTCCATGCCACCCAGGCC AGGAGTGACCAGGAGCCGCTGTACCCAGTGCAGGTCAGCTCTGCGGACGCTCGGCTC ATGGTCTTTGACAAGACGGAAGGGACGTGGCGGCTGCTGTGCTCCTCGCGCTCCAAC GCCAGGGTAGCCGGACTCAGCTGCGAGGAGATGGGCTTCCTCAGGGCACTGACCCAC TCCGAGCTGGACGTGCGAACGGCGGGCGCCAATGGCACGTCGGGCTTCTTCTGTGTG GACGAGGGGAGGCTGCCCCACACCCAGAGGCTGCTGGAGGTCATCTCCGTGTGTGAT TGCCCCAGAGGCCGTTTCTTGGCCGCCATCTGCCAAGACTGTGGCCGCAGGAAGCTG CCCGTGGACCGCATCGTGGGAGGCCGGGACACCAGCTTGGGCCGGTGGCCGTGGCAA GTCAGCCTTCGCTATGATGGAGCACACCTCTGTGGGGGATCCCTGCTCTCCGGGGAC TGGGTGCTGACAGCCGCCCACTGCTTCCCGGAGCGGAACCGGGTCCTGTCCCGATGG CGAGTGTTTGCCGGTGCCGTGGCCCAGGCCTCTCCCCACGGTCTGCAGCTGGGGGTG CAGGCTGTGGTCTACCACGGGGGCTATCTTCCCTTTCGGGACCCCAACAGCGAGGAG AACAGCAACGATATTGCCCTGGTCCACCTCTCCAGTCCCCTGCCCCTCACAGAATACAttorney Docket No.: 49082-0007WO1 ATCCAGCCTGTGTGCCTCCCAGCTGCCGGCCAGGCCCTGGTGGATGGCAAGATCTGT ACCGTGACGGGCTGGGGCAACACGCAGTACTATGGCCAACAGGCCGGGGTACTCCAG GAGGCTCGAGTCCCCATAATCAGCAATGATGTCTGCAATGGCGCTGACTTCTATGGA AACCAGATCAAGCCCAAGATGTTCTGTGCTGGCTACCCCGAGGGTGGCATTGATGCC TGCCAGGGCGACAGCGGTGGTCCCTTTGTGTGTGAGGACAGCATCTCTCGGACGCCA CGTTGGCGGCTGTGTGGCATTGTGAGTTGGGGCACTGGCTGTGCCCTGGCCCAGAAG CCAGGCGTCTACACCAAAGTCAGTGACTTCCGGGAGTGGATCTTCCAGGCCATAAAG ACTCACTCCGAAGCCAGCGGCATGGTGACCCAGCTCTGA SEQ ID NO:37 Hepsin Protein Sequence in vector pFCS- newhHepsin-WPRE-SIN (new ori) 1008-146, CI01.0004 MMSFVSLLLVGILFHATQARSDQEPLYPVQVSSADARLMVFDKTEGTWRLLCSSRSN ARVAGLSCEEMGFLRALTHSELDVRTAGANGTSGFFCVDEGRLPHTQRLLEVISVCD CPRGRFLAAICQDCGRRKLPVDRIVGGRDTSLGRWPWQVSLRYDGAHLCGGSLLSGD WVLTAAHCFPERNRVLSRWRVFAGAVAQASPHGLQLGVQAVVYHGGYLPFRDPNSEE NSNDIALVHLSSPLPLTEYIQPVCLPAAGQALVDGKICTVTGWGNTQYYGQQAGVLQ EARVPIISNDVCNGADFYGNQIKPKMFCAGYPEGGIDACQGDSGGPFVCEDSISRTP RWRLCGIVSWGTGCALAQKPGVYTKVSDFREWIFQAIKTHSEASGMVTQL SEQ ID NO:38 HGF CDS in vector pFCS-SPhHGF-WPRE-SIN (new ori) 1008-146, CI02.0002 ATGATGTCCTTTGTCTCTCTGCTCCTGGTTGGCATCCTATTCCATGCCACCCAGGCC CAAAGGAAAAGAAGAAATACAATTCATGAATTCAAAAAATCAGCAAAGACTACCCTA ATCAAAATAGATCCAGCACTGAAGATAAAAACCAAAAAAGTGAATACTGCAGACCAA TGTGCTAATAGATGTACTAGGAATAAAGGACTTCCATTCACTTGCAAGGCTTTTGTT TTTGATAAAGCAAGAAAACAATGCCTCTGGTTCCCCTTCAATAGCATGTCAAGTGGA GTGAAAAAAGAATTTGGCCATGAATTTGACCTCTATGAAAACAAAGACTACATTAGA AACTGCATCATTGGTAAAGGACGCAGCTACAAGGGAACAGTATCTATCACTAAGAGT GGCATCAAATGTCAGCCCTGGAGTTCCATGATACCACACGAACACAGCTATCGGGGT AAAGACCTACAGGAAAACTACTGTCGAAATCCTCGAGGGGAAGAAGGGGGACCCTGG TGTTTCACAAGCAATCCAGAGGTACGCTACGAAGTCTGTGACATTCCTCAGTGTTCA GAAGTTGAATGCATGACCTGCAATGGGGAGAGTTATCGAGGTCTCATGGATCATACA GAATCAGGCAAGATTTGTCAGCGCTGGGATCATCAGACACCACACCGGCACAAATTC TTGCCTGAAAGATATCCCGACAAGGGCTTTGATGATAATTATTGCCGCAATCCCGAT GGCCAGCCGAGGCCATGGTGCTATACTCTTGACCCTCACACCCGCTGGGAGTACTGT GCAATTAAAACATGCGCTGACAATACTATGAATGACACTGATGTTCCTTTGGAAACA ACTGAATGCATCCAAGGTCAAGGAGAAGGCTACAGGGGCACTGTCAATACCATTTGG AATGGAATTCCATGTCAGCGTTGGGATTCTCAGTATCCTCACGAGCATGACATGACT CCTGAAAATTTCAAGTGCAAGGACCTACGAGAAAATTACTGCCGAAATCCAGATGGG TCTGAATCACCCTGGTGTTTTACCACTGATCCAAACATCCGAGTTGGCTACTGCTCC CAAATTCCAAACTGTGATATGTCACATGGACAAGATTGTTATCGTGGGAATGGCAAA AATTATATGGGCAACTTATCCCAAACAAGATCTGGACTAACATGTTCAATGTGGGAC AAGAACATGGAAGACTTACATCGTCATATCTTCTGGGAACCAGATGCAAGTAAGCTG AATGAGAATTACTGCCGAAATCCAGATGATGATGCTCATGGACCCTGGTGCTACACG GGAAATCCACTCATTCCTTGGGATTATTGCCCTATTTCTCGTTGTGAAGGTGATACC ACACCTACAATAGTCAATTTAGACCATCCCGTAATATCTTGTGCCAAAACGAAACAA TTGCGAGTTGTAAATGGGATTCCAACACGAACAAACATAGGATGGATGGTTAGTTTG AGATACAGAAATAAACATATCTGCGGAGGATCATTGATAAAGGAGAGTTGGGTTCTT ACTGCACGACAGTGTTTCCCTTCTCGAGACTTGAAAGATTATGAAGCTTGGCTTGGA ATTCATGATGTCCACGGAAGAGGAGATGAGAAATGCAAACAGGTTCTCAATGTTTCC CAGCTGGTATATGGCCCTGAAGGATCAGATCTGGTTTTAATGAAGCTTGCCAGGCCTAttorney Docket No.: 49082-0007WO1 GCTGTCCTGGATGATTTTGTTAGTACGATTGATTTACCTAATTATGGATGCACAATT CCTGAAAAGACCAGTTGCAGTGTTTATGGCTGGGGCTACACTGGATTGATCAACTAT GATGGCCTATTACGAGTGGCACATCTCTATATAATGGGAAATGAGAAATGCAGCCAG CATCATCGAGGGAAGGTGACTCTGAATGAGTCTGAAATATGTGCTGGGGCTGAAAAG ATTGGATCAGGACCATGTGAGGGGGATTATGGTGGCCCACTTGTTTGTGAGCAACAT AAAATGAGAATGGTTCTTGGTGTCATTGTTCCTGGTCGTGGATGTGCCATTCCAAAT CGTCCTGGTATTTTTGTCCGAGTAGCATATTATGCAAAATGGATACACAAAATTATT TTAACATATAAGGTACCACAGTCATAG SEQ ID NO: 39 HGF Protein Sequences in vector pFCS- SPhHGF-WPRE-SIN (new ori) 1008-146, CI02.0002 MMSFVSLLLVGILFHATQAQRKRRNTIHEFKKSAKTTLIKIDPALKIKTKKVNTADQ CANRCTRNKGLPFTCKAFVFDKARKQCLWFPFNSMSSGVKKEFGHEFDLYENKDYIR NCIIGKGRSYKGTVSITKSGIKCQPWSSMIPHEHSYRGKDLQENYCRNPRGEEGGPW CFTSNPEVRYEVCDIPQCSEVECMTCNGESYRGLMDHTESGKICQRWDHQTPHRHKF LPERYPDKGFDDNYCRNPDGQPRPWCYTLDPHTRWEYCAIKTCADNTMNDTDVPLET TECIQGQGEGYRGTVNTIWNGIPCQRWDSQYPHEHDMTPENFKCKDLRENYCRNPDG SESPWCFTTDPNIRVGYCSQIPNCDMSHGQDCYRGNGKNYMGNLSQTRSGLTCSMWD KNMEDLHRHIFWEPDASKLNENYCRNPDDDAHGPWCYTGNPLIPWDYCPISRCEGDT TPTIVNLDHPVISCAKTKQLRVVNGIPTRTNIGWMVSLRYRNKHICGGSLIKESWVL TARQCFPSRDLKDYEAWLGIHDVHGRGDEKCKQVLNVSQLVYGPEGSDLVLMKLARP AVLDDFVSTIDLPNYGCTIPEKTSCSVYGWGYTGLINYDGLLRVAHLYIMGNEKCSQ HHRGKVTLNESEICAGAEKIGSGPCEGDYGGPLVCEQHKMRMVLGVIVPGRGCAIPN RPGIFVRVAYYAKWIHKIILTYKVPQS OTHER EMBODIMENTS It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

Attorney Docket No.: 49082-0007WO1 WHAT IS CLAIMED IS:

1. A cell engineered to express a hepatocyte growth factor (HGF) polypeptide or biologically active variant thereof and a hepsin polypeptide or enzymatically active variant thereof.

2. A population of cells engineered to express a hepatocyte growth factor (HGF) polypeptide or biologically active variant thereof and a hepsin polypeptide or enzymatically active variant thereof.

3. A cell comprising one or more polynucleotide sequence(s) encoding a hepatocyte growth factor (HGF) polypeptide or biologically active variant thereof operably linked to an exogenous promoter; and one or more polynucleotide sequence(s) encoding a hepsin polypeptide or enzymatically active variant thereof operably linked to an exogenous promoter.

4. A population of cells comprising one or more polynucleotide sequence(s) encoding a hepatocyte growth factor (HGF) polypeptide or biologically active variant thereof operably linked to an exogenous promoter; and one or more polynucleotide sequence(s) encoding a hepsin polypeptide or enzymatically active variant thereof operably linked to an exogenous promoter.

5. The cell of claim 3 or population of cells of claim 4, wherein the promoter is selected from the group consisting of human CMV, simian CMV, EF1a, SV40, human PGK1, mouse PGK1, Ubc, Human beta actin, CAG, TRE, UAS, CaMKIIa, H1, RSV, UB, and U6.

6. The cell of claim 3 or population of cells of claim 4, wherein the promoter is a cytomegalovirus immediate-early enhancer / promoter.

7. The cell of any one of claims 1, 3, 5, or 6, or population of cells of any one of claims 2, 4, 5, or 6, wherein the HGF polypeptide or biologically active variant thereof comprises a hepsin cleavage amino acid motif.Attorney Docket No.: 49082-0007WO1 8. The cell or population of cells of claim 7, wherein the hepsin cleavage amino acid motif is selected from the group consisting of KKNR, KKTR, KKLR, KQNR, KQTR, KQLR, PQNR, PQTR, PQLR, PKNR, PKTR and PKLR.

9. The cell of any one of claims 1, 3, or 5–8, or the population of cells of any one of claims 2, or 4–8, wherein the HGF polypeptide or biologically active variant thereof comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:

32.

10. The cell of any one of claims 1, 3, or 5–9 or the population of cells of any one of claims 2, or 4–9, wherein the HGF polypeptide or biologically active variant thereof comprises a signal sequence.

11. The cell or population of cells of claim 10, wherein the signal sequence is a native HGF signal sequence.

12. The cell or population of cells of claim 11, wherein the signal sequence is not a native HGF signal sequence.

13. The cell or population of cells of claim 10, wherein the signal sequence is selected from the group consisting of SEQ ID NOs:40–57.

14. The cell of any one of claims 1, 3, or 5–13 or the population of cells of any one of claims 2 or 4–13, wherein the hepsin polypeptide or enzymatically active variant thereof comprises an amino sequence having at least 80% identity to SEQ ID NO:

29.

15. The cell of any one of claims 1, 3, or 5–14 or the population of cells of any one of claims 2 or 4–14, wherein hepsin polypeptide or enzymatically active variant thereof comprises a signal sequence.

16. The cell or population of cells of claim 15, wherein the signal sequence is selected from the group consisting of SEQ ID NOS:40–57.Attorney Docket No.: 49082-0007WO1 17. The cell of any one of claims 1, 3, or 5–16, wherein the cell is a mammalian cell.

18. The cell of claim 17, wherein the cell is a CHO cell.

19. The population of cells of any one of claims 2 or 4–16, wherein the population of cells comprises or consists of mammalian cells.

20. The population of cells of claim 19, wherein the population of cells comprises or consists of CHO cells.

21. The cell of any one of claims 1 or 3–18, wherein the cell comprises polynucleotide sequence(s) encoding a ratio of from 1:1 to 1:1000 copies of the hepsin polypeptide or enzymatically active variant thereof to the HGF polypeptide or biologically active variant thereof.

22. The cell of claim 21, wherein the ratio is at least 50 copies of the HGF polypeptide or enzymatically active variant thereof to 1 of the hepsin polypeptide or biologically active variant thereof.

23. The cell of claim 22, wherein the ratio is from 1:50 to 1:1000.

24. The cell of claim 23, wherein the ratio is from 1:50 to 1:

100.

25. The cell of claim 24, wherein the ratio is from 1:79 to 1:127, optionally about 1:

100.

26. The population of cells of any one of claims 2, 4–17, 19, or 20, wherein the population of cells cell comprises polynucleotide sequence(s) encoding a ratio of from 1:1 to 1:1000 copies of the hepsin polypeptide or enzymatically active variant thereof to the HGF polypeptide or biologically active variant thereof.

27. The population of cells of claim 26, wherein the ratio is at least 50 copies of the HGF polypeptide or enzymatically active variant thereof to 1 of the hepsin polypeptide or biologically active variant thereof.Attorney Docket No.: 49082-0007WO1 28. The population of cells of claim 27, wherein the ratio is about 1:

100.

29. A composition comprising: a first nucleic acid expression construct comprising a polynucleotide sequence encoding hepatocyte growth factor (HGF) or a biologically active variant thereof; and a second nucleic acid expression construct comprising a polynucleotide sequence encoding hepsin or an enzymatically active variant thereof.

30. The composition of claim 29, wherein the HGF polypeptide or biologically active variant thereof comprises a hepsin cleavage amino acid motif.

31. The composition of claim 30, wherein the hepsin cleavage amino acid motif is selected from the group consisting of KKNR, KKTR, KKLR, KQNR, KQTR, KQLR, PQNR, PQTR, PQLR, PKNR, PKTR and PKLR.

32. The composition of any one of claims 29–31, wherein the HGF polypeptide or biologically active variant thereof comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:

32.

33. The composition of any one of claims 29–32, wherein the HGF polypeptide or biologically active variant thereof comprises a signal sequence.

34. The composition of claim 33, wherein the signal sequence is a native HGF signal sequence.

35. The composition of claim 33, wherein the signal sequence is not a native HGF signal sequence.

36. The composition of claim 35, wherein the signal sequence is selected from the group consisting of SEQ ID NOs:40–57.

37. The composition of any one of claims 29–36, wherein the hepsin polypeptide or enzymatically active variant thereof comprises an amino sequence having at least 80% identity to SEQ ID NO: 29.Attorney Docket No.: 49082-0007WO1 38. The composition of any one of claims 29–37, wherein hepsin polypeptide or enzymatically active variant thereof comprises a signal sequence.

39. The composition of claim 38, wherein the signal sequence is selected from the group consisting of SEQ ID NOS: 40–57.

40. The composition of any one of claims 29–39, wherein the first nucleic acid expression construct further comprises a nucleic acid sequence encoding a promoter operably linked to the polynucleotide encoding HGF.

41. The composition of claim 40 wherein the promoter is selected from the group consisting of human CMV, simian CMV, EF1a, SV40, human PGK1, mouse PGK1, Ubc, Human beta actin, CAG, TRE, UAS, CaMKIIa, H1, RSV, UB, and U6.

42. The composition of claim 40, wherein the promoter is a cytomegalovirus immediate-early enhancer / promoter.

43. The composition of any one of claims 29–42, wherein the second nucleic acid expression construct further comprises a nucleic acid sequence encoding a promoter, operably linked to the polynucleotide encoding hepsin or an enzymatically active variant thereof.

44. The composition of claim 43, wherein the promoter is selected from the group consisting of human CMV, simian CMV, EF1a, SV40, human PGK1, mouse PGK1, Ubc, Human beta actin, CAG, TRE, UAS, CaMKIIa, H1, RSV, UB, and U6.

45. The composition of claim 43, wherein the promoter is a cytomegalovirus immediate-early enhancer / promoter.

46. The composition of any one of claims 29–45, wherein the first and / or second nucleic acid expression construct(s) each, independently, further comprise one or more of: a nucleic acid sequence encoding an attachment site, a nucleic acid sequence encoding a selectable marker, a nucleic acid sequence encoding a posttranscriptional regulatory element, and a poly A signal sequence.Attorney Docket No.: 49082-0007WO1 47. A cell comprising the composition of any one of claims 29–46.

48. A cell transformed, transfected, or transduced with the composition of any one of claims 29–46.

49. A population of cells transformed, transfected, or transduced with the composition of any one of claims 29–46.

50. The cell according to claim 47 or 48 or population of cells according to claim 49, wherein the cell or population of cells is transformed, transfected, or transduced with a plurality of first and second nucleic acid expression constructs in a ratio of from 1:1 to 1:1000 of the second (hepsin) construct to the first (HGF) construct.

51. The cell or population of cells according to claim 50, wherein the cell or population of cells is transformed, transfected, or transduced with a plurality of first and second nucleic acid expression constructs in a ratio of from 1:10 to 1:1000 of the second (hepsin) construct to the first (HGF) construct.

52. The cell or population of cells according to claim 51, wherein cell or population of cells is transformed, transfected, or transduced with a plurality of first and second nucleic acid constructs in a ratio of about 1:

100.

53. The cell according to any one of claims 47, 48, or 50–52 or population of cells according to any one of claims 49–52, wherein the cell(s) are mammalian cell.

54. The cell or population of cells of claim 53, wherein the cell(s) are CHO cell(s).

55. A method of producing activated HGF, the method comprising: culturing the cell of any one of claims 1, 3, 5–18, 21–25, 47, 48, or 50–54 or the population of cells of any one of claims 2, 4–16, 19, 20, 26–28, or 49–54 under conditions suitable for a) expression of the HGF or biologically active variant thereof and of the hepsin or enzymatically active portion thereof; and b) cleavage of the HGF or biologically active variant thereof with the hepsin or enzymatically active portion thereof, thereby producing activated HGF.Attorney Docket No.: 49082-0007WO1 56. The method of claim 55, further comprising: isolating and / or purifying the activated HGF.

57. The method of claim 55 or claim 56, wherein the activated HGF comprises a first polypeptide comprising an amino acid sequence having at least 80% identity to SEQ ID NO:33 and a second polypeptide comprising an amino acid sequence having at least 80% identity to SEQ ID NO:

34.

58. The method of claim 57, wherein the first polypeptide and second polypeptide are linked by one or more disulfide bonds.

59. The method of any one of claims 55–58, wherein the activated HGF is capable of binding to c-MET and / or activating the MAPK pathway in epithelial cells.

60. An activated HGF polypeptide produced according to the method of any one of claims 55–59.

61. A method of producing a cell line co-expressing HGF and hepsin, the method comprising: transforming, transducing, or transfecting a population of cells with the composition of any one of claims 29–56.

62. The method of claim 61, wherein the population of cells is transformed, transfected, or transduced with a plurality of first and second nucleic acid expression constructs in a ratio of 1:1 to 1:1000 of the second (hepsin) construct to the first (HGF) construct.

63. The method of claim 62, wherein the population of cells is transformed, transfected, or transduced with a plurality of first and second nucleic acid expression constructs in a ratio of 1:10 to 1:1000 of the second (hepsin) construct to the first (HGF) construct.

64. The method of claim 62, wherein the population of cells is transformed, transfected, or transduced with a plurality of first and second nucleic acid expressionAttorney Docket No.: 49082-0007WO1 constructs in a ratio of about 1:100 of the second (hepsin) construct to the first (HGF) construct.

65. The method of any one of claims 61–64, wherein the cells are mammalian cell.

66. The method of claim 65, wherein the cell(s) are CHO cells.

67. The method of claim any one of claims 61–66, further comprising: isolating a cell from the transformed, transduced, or transfected population of cells.

68. The method of claim 67, wherein the isolated cell comprises polynucleotide sequence(s) encoding a ratio of from 1:1 to 1:1000 copies of the hepsin polypeptide or enzymatically active variant thereof to the HGF polypeptide or biologically active variant thereof.

69. The cell of claim 68, wherein the ratio is at least 50 copies of the HGF polypeptide or enzymatically active variant thereof to 1 of the hepsin polypeptide or biologically active variant thereof.

70. The cell of claim 69, wherein the ratio is from 1:50 to 1:1000.

71. The cell of claim 70, wherein the ratio is from 1:50 to 1:

100.

72. The cell of claim 71, wherein the ratio is from 1:79 to 1:127, optionally about 1:

100.

73. The method of any one of claims 67–72, further comprising: clonally expanding the isolated cell.

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