Methods and compositions

Abstract

Provided herein are recombinant bacterial cells capable of expressing a heterologous extracellular matrix (ECM) degrading polypeptide that degrades an extracellular matrix component. The invention also provides promoters, nucleic acids, and cells comprising the same. The invention also provides medical uses for and methods of using the heterologous bacterial cells, promoters, nucleic acids, and cells provided herein.

Description

[0001] Methods and Compositions

[0002] Field of the invention

[0003] The present invention is in the field of cancer therapy.

[0004] Background

[0005] Immunological cancer therapies including immune checkpoint inhibitors (ICIs), antibody therapies, and engineered cell therapies such as chimeric antigen receptor T-cell (CAR-T) therapies, have changed the treatment landscape of many tumours, proving highly efficacious treatment options for previously intractable cancers.

[0006] Cancer immunotherapies typically require the infiltration of host and / or therapeutically administered cells into the core of a tumour to be effective. However, the extracellular matrix of the tumour can act as a physical barrier to effective immune cell infiltration into the tumour core, contributing to the development of immune excluded tumors.

[0007] Immune-excluded tumours are characterised by a tumour microenvironment (TME) which physically excludes immune cells from the tumour core, thereby limiting the efficacy of cancer immunotherapies. The extracellular matrix (ECM) is a major contributor to the exclusive nature of the microenvironment in immune-excluded tumours, which are a subset of tumours known as "immunologically cold" tumours.

[0008] In a significant number of immune-excluded tumours extracellular matrix (ECM) components are present in elevated levels, presenting a barrier to immunotherapies and drastically decreasing their therapeutic impact. The extracellular matrix (ECM) can mediate exclusion in a number of ways; for example, the extracellular matrix (ECM) makes up a dense, highly fibrotic environment with high levels of interstitial pressure which generates biophysical barriers to the infiltration of immune cells and other biologies; and also traps immunosuppressive growth factors and signals directly into immune cells, affecting their behaviour directly and inhibiting their migration and invasion. Among these elevated extracellular matrix (ECM) components are proteoglycans such as hyaluronic acid (HA) and versican. Most solid tumour indications have been shown to have a percentage of immune-excluded patients. For example, it is estimated that about 50% of patients with triple negative breast cancer carry a tumour that is an immune-excluded tumour. Among the indications with the highest percentage of immune exclusion are triple-negative breast cancer; colon cancer; and pancreatic cancer. These are also indications with a highlyabundant extracellular matrix deposition, which includes increased levels of proteoglycans (for example, hyaluronic acid (HA)).

[0009] It is estimated that worldwide immune-excluded tumours can affect more than 3 million new patients every year, and there are currently no adequate therapies for patients with these tumour types, which also do not respond to immunotherapy strategies. Immune-excluded tumours therefore represent a significant challenge to the effective treatment of a large cohort of cancer patients, contributing unnecessary cancer-associated morbidity and mortality.

[0010] Several avenues have been explored to disrupt the solid tumour stroma in order to remove barriers to infiltration, making immune-excluded tumours accessible to immune cells, and hence more tractable to immunotherapy. For example, some approaches have included using chemotherapeutic agents, radiotherapy, and inflammatory cytokines to potentiate immunotherapy of immune-excluded tumours. However, these approaches do not address the fundamental physical constraints imposed on the tumour by extracellular matrix (ECM); and the untargeted nature of several of these approaches may lead to unwanted side-effects.

[0011] Previous work has shown that using hyaluronidase to degrade hyaluronic acid (HA) in mouse models of cancer leads to normalised vascular coverage, a reduction in hypoxic levels and increased infiltration, without affecting tumour spread (Jacobetz et al., 2013, " Hyaluronan impairs vascular function and drug delivery in a mouse model of pancreatic cancer", Gut, 62(1): 112-120; Clift et al., 2019, " Remodeling the Tumor Microenvironment Sensitizes Breast Tumors to Anti-Programmed Death-Ligand 1 Immunotherapy", Cancer Res., 79(16):4149-4159; each herein incorporated by reference in their entirety). Previous attempts to transfer this to the clinic involved modifying the enzyme through a PEGylation process to increase its time in circulation. This led to toxicity associated with circulating levels of the enzyme, as hyaluronic acid (HA) is produced with high turnover levels in most healthy tissues. In fact, doses administered to patients in clinical trials were up to three orders of magnitude lower than those originally tested in mouse models. Decreasing the dosage of the modified enzyme to reduce toxicity led to reduced efficacy and a Phase III trial was ultimately unsuccessful (Van Cutsem et al., 2020, " Randomized Phase III Trial of Pegvorhyaluronidase Alfa With Nab-Paclitaxel Plus Gemcitabine for Patients With Hyaluronan-High Metastatic Pancreatic Adenocarcinoma", J Clin Oncol., 38(27):3185-3194; herein incorporated by reference in its entirety). Subsequent data shows that the doses administered in patients were ineffective in mouse models, demonstrating that thedose reduction caused by toxicity levels likely led to the lack of efficacy found in clinical trials.

[0012] In International Patent Application No. PCT / GB2024 / 051897, the present inventors provided recombinant cells which are capable of degrading components of the extracellular matrix (ECM) by expressing and optionally secreting extracellular matrix (ECM) degrading polypeptides. By linking the extracellular matrix (ECM) degrading polypeptide genes provided in PCT / GB2024 / 051897 under the control of a hypoxia- or pH-inducible promoter, the inventors were able to direct selective expression of the extracellular matrix (ECM) degrading polypeptide within the tumour microenvironment, which are associated with hypoxia and an acidic pH. Conditions of the tumour microenvironment beyond pH and hypoxia were not considered, however.

[0013] There therefore exists a need to develop improved safe and effective therapeutic agents that break down extracellular matrix (ECM) components of immune-excluded tumours, without affecting the stroma of healthy tissues.

[0014] Brief description of the invention

[0015] The inventors have developed recombinant cells which are capable of degrading components of the extracellular matrix (ECM) by expressing and optionally secreting extracellular matrix (ECM) degrading polypeptides. The inventors have surprisingly found that the cells are capable of degrading the extracellular matrix (ECM), including the extracellular matrix (ECM) of a tumour.

[0016] The inventors have also developed promoters, which surprisingly allow the expression of a gene to which the promoter is operably linked within a specific pH range, hypoxic conditions, or in the presence of N-acetylglucosamine, lactate, and / or ethanolamine. The pH range and hypoxic conditions in which the pH and hypoxia-inducible promoters are active are the same as the pH and hypoxic conditions that are present in the tumour microenvironment; and N-acetylglucosamine, lactate, and ethanolamine are known to be present in the tumour microenvironment. Accordingly, the inventors have surprisingly developed improved promoters that are capable of being activated in the tumour microenvironment, or to put another way, tumour-inducible promoters. By operably linking the tumour-inducible promoters to the extracellular matrix (ECM) degrading polypeptides, the invention allows the expression of the extracellular matrix (ECM) degrading polypeptides and degradation of extracellular matrix (ECM) by said extracellular matrix (ECM) degrading polypeptides may be controlled in response to pH, hypoxia, N-acetylglucosamine, lactate, and / or ethanolamine.The inventors have further surprisingly found that when administered to a subject, the recombinant cells home to and accumulate specifically in tumour tissue. In in vitro experiments, the inventors have found that the recombinant cells infiltrate the tumour tissue, and further increase chimeric antigen receptor T-cell (CAR-T cell) killing of tumour cells within the tissue.

[0017] The inventors have therefore developed recombinant cells that when administered to a subject are capable of specifically localising to and degrading the extracellular matrix (ECM) of a tumour, facilitated improved infiltration of the tumour by therapeutic agents. Accordingly, the inventors have developed recombinant cells that are surprisingly capable of making immune-excluded tumours accessible to therapeutic agents.

[0018] The inventors have also developed nucleic acids, medical methods, medical uses, and therapies relating to the recombinant cells.

[0019] Detailed description of the invention

[0020] The invention is as set out in the claims.

[0021] Recombinant bacterial cells

[0022] In a first aspect, the invention provides a recombinant bacterial cell capable of expressing a heterologous extracellular matrix (ECM) degrading polypeptide that degrades an extracellular matrix component. By "capable of expressing" it is meant that the bacterial cell can express a particular polypeptide - for example an extracellular matrix (ECM) degrading polypeptide - under certain conditions, for example by transcribing messenger RNA (mRNA) from a nucleotide sequence encoding said polypeptide, and / or translating said mRNA into said polypeptide. In some cases, by "capable of expressing" it is meant that the cell does express a particular polypeptide. In some cases, by "capable of expressing" it is meant that the cell expresses a particular polypeptide. Accordingly, in a related aspect, the invention provides a recombinant bacterial cell that expresses a heterologous extracellular matrix (ECM) degrading polypeptide that degrades an extracellular matrix component.

[0023] The heterologous ECM degrading polypeptide may be any ECM degrading polypeptide. In some embodiments the extracellular matrix degrading polypeptide is:

[0024] a) a hyaluronidase, for example a microbial hyaluronidase or a mammalian hyaluronidase;for example wherein the microbial hyaluronidase is a bacterial hyaluronidase;

[0025] b) chondroitin ABC lyase optionally a microbial chondroitin ABC lyase or a mammalian chondroitin ABC lyase;

[0026] for example wherein the microbial chondroitin ABC lyase is a bacterial chondroitin ABC lyase;

[0027] c) neuraminidase, for example a microbial neuraminidase or a mammalian neuraminidase;

[0028] for example wherein the microbial neuraminidase is a bacterial neuraminidase; and / or

[0029] d) PNGase for example a microbial PNGase or a mammalian PNGase;

[0030] for example wherein the microbial PNGase is a bacterial PNGase.

[0031] Various means of making a recombinant bacterial cell capable of expressing a particular polypeptide - for example an extracellular matrix (ECM) degrading polypeptide - are known to the person skilled in the art. For example, a cell may be transformed or transfected with a nucleic acid comprising a nucleotide sequence that encodes the heterologous extracellular matrix degrading (ECM) polypeptide.

[0032] Accordingly, in one embodiment, the cell comprises a nucleic acid comprising a nucleotide sequence that encodes the heterologous extracellular matrix degrading (ECM) polypeptide. The nucleic acid may be any nucleic acid. Accordingly, in some embodiments, the nucleic acid is DNA. Appropriate forms of DNA are known in the art, and are discussed herein. In some embodiments, the nucleic acid is RNA. Appropriate forms of RNA are known in the art, and include mRNA, single-stranded RNA (ssRNA), double-stranded RNA (dsRNA), selfreplicating RNA (srRNA), RNA viral vectors, and the like.

[0033] As discussed herein, the extracellular matrix (ECM) may comprise proteoglycans. Without wishing to be bound by any theory, it is believed that the degradation of the proteoglycan components of extracellular matrix (ECM) comprised in a tumour will allow improved access of therapeutic agents such as immunotherapy agents into the tumour.

[0034] As discussed herein, an extracellular matrix (ECM) component that may suitably be targeted for degradation is a proteoglycan. Suitable ECM degrading polypeptides that are capable of degrading proteoglycans are known in the art, and include (but are not limited to): a hyaluronidase, a chondroitin ABC lyase, a neuraminidase, and / or a PNGase. Hyaluronidase, chondroitin ABC lyase, neuraminidase, and PNGase are broadly distributedthroughout eukaryotes and prokaryotes. Accordingly, the extracellular matrix (ECM) degrading polypeptide may be selected from:

[0035] a) a eukaryotic hyaluronidase, a eukaryotic chondroitin ABC lyase, a eukaryotic neuraminidase, and / or a eukaryotic PNGase; and / or

[0036] b) a prokaryotic hyaluronidase, a prokaryotic chondroitin ABC lyase, a prokaryotic neuraminidase, and / or a prokaryotic PNGase.

[0037] In some embodiments then, the ECM component is a proteoglycan and the extracellular matrix degrading polypeptide is:

[0038] a) a hyaluronidase, for example a microbial hyaluronidase or a mammalian hyaluronidase;

[0039] for example wherein the microbial hyaluronidase is a bacterial hyaluronidase;

[0040] b) chondroitin ABC lyase optionally a microbial chondroitin ABC lyase or a mammalian chondroitin ABC lyase;

[0041] for example wherein the microbial chondroitin ABC lyase is a bacterial chondroitin ABC lyase;

[0042] c) neuraminidase, for example a microbial neuraminidase or a mammalian neuraminidase;

[0043] for example wherein the microbial neuraminidase is a bacterial neuraminidase; and / or

[0044] d) PNGase for example a microbial PNGase or a mammalian PNGase;

[0045] for example wherein the microbial PNGase is a bacterial PNGase.

[0046] The prokaryotic extracellular matrix (ECM) degrading polypeptide may be a microbial extracellular matrix (ECM) degrading polypeptide. The microbial extracellular matrix (ECM) degrading polypeptide may suitably be a bacterial extracellular matrix (ECM) degrading polypeptide. The eukaryotic extracellular matrix (ECM) degrading polypeptide may suitably be a mammalian extracellular matrix (ECM) degrading polypeptide, for example a human extracellular matrix (ECM) degrading polypeptide.

[0047] Accordingly, in some embodiments the extracellular matrix component is a proteoglycan, and:

[0048] a) the ECM degrading polypeptide is a hyaluronidase optionally a microbial hyaluronidase or a mammalian hyaluronidase;

[0049] optionally wherein the microbial hyaluronidase is a bacterial hyaluronidase; b) the ECM degrading polypeptide is chondroitin ABC lyase optionally a microbial chondroitin ABC lyase or a mammalian chondroitin ABC lyase;optionally wherein the microbial chondroitin ABC lyase is a bacterial chondroitin ABC lyase;

[0050] c) the ECM degrading polypeptide is neuraminidase, optionally a microbial neuraminidase or a mammalian neuraminidase;

[0051] optionally wherein the microbial neuraminidase is a bacterial neuraminidase; and / or d) the ECM degrading polypeptide is PNGase optionally a microbial PNGase or a mammalian PNGase;

[0052] optionally wherein the microbial PNGase is a bacterial PNGase.

[0053] As discussed herein, extracellular matrix (ECM) components that are proteoglycans and which may suitably be targeted for degradation include (but are not limited to) hyaluronic acid (HA) and versican.

[0054] Hyaluronic acid (HA) is a an anionic, nonsulfated glycosaminoglycan distributed widely throughout connective, epithelial, and neural tissues; and is a major component of the extracellular matrix, contributing to cell proliferation and migration; and the progression of tumors. Hyaluronic acid (HA) has the following structure:

[0055]

[0056] Where n is an integer representing the number of monomers in a polymer of hyaluronic acid. As used herein, the terms "hyaluronic acid", "hyaluronan", and " HA" are interchangeable.

[0057] Versican is a large extracellular matrix chondroitin sulfate proteoglycan present in a variety of human tissues, and is encoded by the VCAN gene. Versican is made up of a central protein core bound by multiple chondroitin sulfate chains, and it has 5 different isoforms created by differential or alternative splicing. Versican is involved in cell adhesion, migration, and proliferation; and increased versican expression is associated with tumour growth and metastasis.

[0058] As is known in the art, hyaluronic acid (HA) may be degraded by a hyaluronidase. Versican may be degraded by different classes of enzymes which target different parts of the glycanchains, including sialic acid, glycosaminoglycans (GAGs), and glycoside bonds. These include a chondroitin ABC lyase, a neuraminidase, and / or PNGase.

[0059] As used herein, the term "degrading", "degraded", or "degrade" relates to the process of breaking down a polymer into its constituent subunits, and may include breaking down a polymer chain of a first length into two polymer chains each of a second, shorter length; and / or the breaking down a polymer chain into its constituent monomers or fragments thereof.

[0060] Accordingly, in some embodiments:

[0061] a) the ECM component is hyaluronic acid (HA) and the ECM degrading polypeptide is a hyaluronidase, optionally a microbial hyaluronidase or a mammalian hyaluronidase; optionally wherein the microbial hyaluronidase is a bacterial hyaluronidase; b) the ECM component is Versican and the ECM degrading polypeptide is chondroitin ABC lyase optionally a microbial chondroitin ABC lyase or a mammalian chondroitin ABC lyase;

[0062] optionally wherein the microbial chondroitin ABC lyase is a bacterial chondroitin ABC lyase;

[0063] c) the ECM component is Versican and the ECM degrading polypeptide is neuraminidase, optionally a microbial neuraminidase or a mammalian neuraminidase; optionally wherein the microbial neuraminidase is a bacterial neuraminidase; and / or d) the ECM component is Versican and the ECM degrading polypeptide is PNGase optionally a microbial PNGase or a mammalian PNGase;

[0064] optionally wherein the microbial PNGase is a bacterial PNGase.

[0065] As discussed herein, the extracellular matrix (ECM) degrading polypeptide may be any suitable extracellular matrix (ECM) degrading polypeptide, for example any suitable hyaluronidase, chondroitin ABC lyase, neuraminidase, and / or PNGase. In the context of degrading an extracellular matrix (ECM) component in a tumour, it will be understood that an extracellular matrix (ECM) degrading polypeptide (for example, a hyaluronidase, chondroitin ABC lyase, neuraminidase, and / or PNGase) is "suitable" if it is capable of or degrades extracellular matrix (ECM) components in a tumour environment or microenvironment. As is known in the art, a tumour environment or microenvironment may be hypoxic, anaerobic, anoxic, or microaerobic. For example, a tumour environment or microenvironment may have an oxygen concentration of below 2%, for example 0-2% and / or 1-2% (McKeown, 2014, " Defining normoxia, physioxia and hypoxia in tumours -implications for treatment response", Br. J. Radiol., 87(1035):20130676). A tumour environment or microenvironment may be acidic, for example may have a pH of pH 6.2 ±0.25 (Chien et al., 2022, " Enhancing the tropism of bacteria via genetically programmed biosensors", Nat. Biomed. Eng., 6:94-104; the contents of which are herein incorporated by reference in their entirety)). Methods of determining whether an extracellular matrix (ECM) degrading polypeptide is capable of degrading extracellular matrix (ECM) in hypoxic, anaerobic, anoxic, microaerobic, and / or acidic conditions are known in the art, and may be found - for example - in Oueslati et al. 2014, " CTAB turbidimetric method for assaying hyaluronic acid in complex environments and under cross-linked form", Carbohydrate polymers, 112:102-108 (herein incorporated by reference in its entirety).

[0066] In some embodiments, the heterologous extracellular matrix (ECM) degrading polypeptide: a) is a hyaluronidase and comprises or consists of an amino acid sequence of SEQ ID NO: 1; is a chondroitin ABC Lyase and comprises or consists of an amino acid sequence of SEQ ID NO: 3 or 47; is a neuraminidase and comprises or consists of an amino acid sequence of SEQ ID NO: 5; or is a PNGase and comprises or consists of an amino acid sequence of SEQ ID NO: 7; or comprises or consists of an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, or SEQ ID NO: 47; and / or

[0067] b) is encoded by a nucleotide sequence of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, or SEQ ID NO: 48 or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, or SEQ ID NO: 48.

[0068] In some embodiments, the heterologous extracellular matrix (ECM) degrading polypeptide is a bacterial hyaluronidase; optionally

[0069] wherein the heterologous extracellular matrix (ECM) degrading polypeptide: a) comprises or consists of an amino acid sequence of SEQ ID NO: 1; or of an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 1; and / or

[0070] b) is encoded by a nucleotide sequence of SEQ ID NO: 2, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 2.

[0071] The term "identical to" is known to the person skilled in the art, and relates to the percentage identity between two or more nucleic acid sequences or two or more aminoacid sequences. The terms "identical to" and "sequence identity" are interchangeable -i.e., where a first sequence is X% (where X is an integer between 0 and 100) "identical to" a second sequence, the first sequence shares X% "sequence identity" with the second sequence.

[0072] To determine the percentage identity of two amino acid sequences or of two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in the sequence of a first amino acid or nucleic acid sequence for optimal alignment with a second amino acid or nucleic acid sequence). The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences (i.e., % identity=number of identical overlapping positions / total number of positions×100%). In one embodiment, the two sequences are the same length.

[0073] Percentage identity between two sequences can also be identified using a mathematical algorithm. One example of a mathematical algorithm for the comparison of two sequences is the algorithm of Karlin and Altschul, 1990, Proc. Natl. Acad. Sci. U. S. A. 87:2264 2268 (herein incorporated by reference in its entirety), modified as in Karlin and Altschul, 1993, Proc. Natl. Acad. Sci. U. S. A. 90:5873 5877 (herein incorporated by reference in its entirety). Such an algorithm is incorporated into the NBLAST and XBLAST programs of Altschul etal., 1990, J. Mol. Biol. 215:403 (herein incorporated by reference in its entirety). To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al., 1997, Nucleic Acids Res. 25:3389 3402 (herein incorporated by reference in its entirety). Alternatively, PSI BLAST can be used to perform an iterated search which detects distant relationships between molecules (Id.). When utilizing BLAST, Gapped BLAST, and PSI Blast programs, the default parameters of the respective programs (e.g., of XBLAST and NBLAST) can be used (see, e.g., National Center for Biotechnology Information (NCBI) on the worldwide web, ncbi.nlm.nih.gov). Another example of a mathematical algorithm for the comparison of sequences is the algorithm of Myers and Miller, 1988, CABIOS 4:11 17 (herein incorporated by reference in its entirety). Such an algorithm is incorporated in the ALIGN program (version 2.0) which is part of the GCG sequence alignment software package. Another example of a mathematical algorithm for the comparison of two sequences is the algorithm of Thompson et al., 1994, Nucleic Acids Res. 22(22):4673-80 (herein incorporated by reference in its entirety), incorporated into the ClustalW program.The percentage identity between two sequences can be determined using techniques similar to those described above, with or without allowing gaps. In calculating percentage identity, typically only exact matches are counted.

[0074] In some embodiments, the extracellular matrix (ECM) degrading polypeptide has hyaluronate lyase activity and / or is a hyaluronate lyase. In some embodiments, hyaluronidase has hyaluronate lyase activity and / or is a hyaluronate lyase. Enzymes with hyaluronate lyase activity and / or that are a hyaluronate lyase catalyse the cleavage of hyaluronate chains at a 0-D-GalNAc-(l— >4)-[3-D-GlcA bond, ultimately breaking the polysaccharide down to 3-(4-deoxy-[3-D-gluc-4-enuronosyl)-N-acetyl-D-glucosamine. Hyaluronate lyases are typically produced by bacterial cells and are not produced by eukaryotic cells. Accordingly, in some embodiments, the extracellular matrix (ECM) degrading polypeptide is a bacterial hyaluronidase, wherein the bacterial hyaluronidase has hyaluronate lyase activity and / or is a hyaluronate lyase.

[0075] In some embodiments, the extracellular matrix (ECM) degrading polypeptide is hyaluronate lyase EC 4.2.2.1 (ExplorEnz - the Enzyme Database - EC 4.2.2.1; available at https: / / www.enzyme-database.org / query.php?ec=4.2.2.1). In some embodiments, the hyaluronidase is hyaluronate lyase EC 4.2.2.1 (ExplorEnz - the Enzyme Database - EC 4.2.2.1; available at https: / / www.enzyme-database.org / query.php?ec=4.2.2.1). In some embodiments, the hyaluronate lyase:

[0076] a) comprises or consists of an amino acid sequence of SEQ ID NO: 1; or of an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 1; and / or

[0077] b) is encoded by a nucleotide sequence of SEQ ID NO: 2, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 2.

[0078] In some embodiments, the extracellular matrix (ECM) degrading polypeptide does not have glycoside hydrolase activity and / or is not a glycoside hydrolase. In some embodiments, the hyaluronidase does not have glycoside hydrolase activity and / or is not a glycoside hydrolase. Enzymes with glycoside hydrolase activity that are active on hyaluronate include hyaluronoglucosaminidase, which catalyses the random hydrolysis of (l->4)-linkages between N-acetyl-beta-D-glucosamine and D-glucuronate residues in hyaluronate; and hyaluronoglucuronidase, which catalyses the random hydrolysis of (1->3)-linkages between beta-D-glucuronate and N-acetyl-D-glucosamine residues inhyaluronate. Hyaluronidases with glycoside hydrolase activity are typically produced by eukaryotic cells and are not produced by bacterial cells. Accordingly, in some embodiments, the extracellular matrix (ECM) degrading polypeptide is not a eukaryotic hyaluronidase, wherein the eukaryotic hyaluronidase has glycoside hydrolase activity and / or is a glycoside hydrolase.

[0079] In some embodiments, the extracellular matrix (ECM) degrading polypeptide is not hyaluronoglucosaminidase. In some embodiments, the hyaluronidase is not hyaluronoglucosaminidase. In some embodiments, the extracellular matrix (ECM) degrading polypeptide is not hyaluronoglucosaminidase EC 3.2.1.35 (Expasy - ENZYME entry: EC 3.2.1.35; available at: https: / / purl.expasy. Org / enzyme / EC / 3.2.l.35). In some embodiments, the hyaluronidase is not hyaluronoglucosaminidase EC 3.2.1.35 (Expasy -ENZYME entry: EC 3.2.1.35; available at:

[0080]

[0081] h Ups; / / purL expasy^ In some embodiments, the extracellular matrix (ECM) degrading polypeptide is not hyaluronoglucuronidase. In some embodiments, the hyaluronidase is not hyaluronoglucuronidase. In some embodiments, the extracellular matrix (ECM) degrading polypeptide is not hyaluronoglucuronidase EC 3.2.1.36 (Expasy - ENZYME entry: EC 3.2.1.36; available at: https: / / enzyme.expasy. Org / EC / 3.2.l.36). In some embodiments, the hyaluronidase is not hyaluronoglucuronidase EC 3.2.1.36 (Expasy - ENZYME entry: EC 3.2.1.36; available at: https: / / enzyme.expasy. Org / EC / 3.2.l.36).

[0082] In some embodiments, the extracellular matrix (ECM) degrading polypeptide is not: a) a hymenopteran hyaluronidase; optionally a bee venom hyaluronidase, a wasp venom hyaluronidase, or a tiger wasp venom hyaluronidase;

[0083] b) a human hyaluronidase; optionally human hyaluronidase-1 (hHyal-1), human hyaluronidase-2 (hHyal-2), human PH-20 (hPH-20), or bovine testes hyaluronidase (BTS); c) a Streptomyces koganeiensis hyaluronidase, a Streptomyces zooepidemic hyaluronidase, or a Streptomyces pristinaespiralis hyaluronidase;

[0084] d) a Streptococcus zooepidemicus hyaluronidase; or

[0085] e) a Penicillium funiculosum hyaluronidase;

[0086] f) an annelid hyaluronidase; optionally a leech hyaluronidase.

[0087] Bacterial hyaluronidase has an optimum activity in conditions between pH 5.5 and pH 6. Accordingly, in some embodiments, the hyaluronidase has optimum activity pH between pH 5.5 and pH 6. In some embodiments, the hyaluronidase is a bacterial hyaluronidase that has optimum activity pH between pH 5.5 and pH 6.

[0088] Human hyaluronidases have an optimum activity between pH 3 and pH 4. Accordingly, in some embodiments, the hyaluronidase does not have optimum activity between pH 3 and pH 4.Leech hyaluronidases have an optimum activity at about pH 7, for example have an optimum activity at pH 7. Accordingly, in some embodiments, the hyaluronidase does not have an optimum activity at about pH 7. In some embodiments, the hyaluronidase does not have an optimum activity at pH 7.

[0089] Methods of determining the pH for the optimum activity of an enzyme are known to the person skilled in the art and typically include testing the activity of an enzyme at a range of pH values to identify the pH at which the activity (for example, catalytic rate) of the enzyme is highest. Suitably, the turbidimetric assay described herein may be used to determine the pH at which a hyaluronidase has optimum activity. In the turbidimetric assay, the pH at which a hyaluronidase has optimum activity would be the pH condition lowest OD600at a given time point, and / or the pH condition at which the OD600reaches a pre-determined value at the earliest timepoint ( / .e., the fastest), compared to other pH conditions tested.

[0090] In some embodiments, the extracellular matrix (ECM) degrading polypeptide is not: a) an influenza neuraminidase, optionally an avian influenza neuraminidase;

[0091] b) a Clostridium perfringens neuraminidase, optionally Nani or NanJ from Clostridium perfringens;

[0092] c) a Chinese hamster ovary (CHO) cell sialidase or a Chinese hamster ovary (CHO) cell neuraminidase;

[0093] d) a Bacteroides fragilis neuraminidase, optionally NanH from Bacteroides fragilis; or e) a Streptococcus pneumoniae neuraminidase.

[0094] As will be appreciated, for a gene to be expressed from a nucleic acid, it may be advantageous or required to incorporate a promoter into the nucleic acid. Accordingly, in some embodiments, the nucleic acid further comprises a promoter. The term "promoter" is known to the person skilled in the art, and is typically a portion of a nucleic acid comprising a nucleotide sequence to which proteins (for example, sigma factors, transcription factors, and RNA polymerase) bind in order to initiate or drive transcription of a single nucleic acid (for example, RNA or mRNA) transcript from the nucleic acid (for example, DNA) downstream of the promoter. In some embodiments, the nucleotide sequence encoding the heterologous extracellular matrix (ECM) degrading polypeptide is operably linked to the promoter. By "operably linked", it is meant that the promoter is capable of initiating or driving transcription of a single nucleic acid transcript (for example, RNA or mRNA) comprising a nucleotide sequence encoding the heterologous extracellular matrix (ECM) degrading polypeptide from the nucleic acid (for example, DNA).A promoter may be a constitutive promoter or may be an inducible promoter. The term "constitutive promoter" and "inducible promoter" are known to the person skilled in the art. In some instances, it may be advantageous to control expression of gene encoding a heterologous extracellular matrix (ECM) degrading polypeptide or transcription of a single nucleic acid transcript (for example, RNA or mRNA) comprising a nucleotide sequence encoding the heterologous extracellular matrix (ECM) polypeptide from the nucleic acid (for example, DNA). As will be appreciated, this may be achieved using an inducible promoter from which gene expression or single nucleic acid transcription is initiated or driven in response to the presence or concentration of a particular molecule, or in response to particular environmental conditions such as pH, oxygen concentration, and / or temperature. Accordingly, in some embodiments, the promoter is an inducible promoter.

[0095] In particular, it may be advantageous to restrict expression of the heterologous extracellular matrix (ECM) degrading polypeptide by the cell to the tumour environment or tumour microenvironment. Accordingly, in some embodiments, the inducible promoter is a tumour-inducible promoter. As discussed herein, the tumour environment or tumour microenvironment may be anaerobic, anoxic, or microaerobic; and / or may have an exemplary oxygen concentration of below 2%, for example between 0-2% and / or between 1-2% (McKeown, 2014, " Defining normoxia, physioxia and hypoxia in tumours -implications for treatment response", Br. J. Radiol., 87(1035):20130676; herein incorporated by reference in its entirety). A tumour environment or microenvironment may be acidic, for example may have a pH of pH 6.2 ± 0.25 (Chien et al., 2022, " Enhancing the tropism of bacteria via genetically programmed biosensors", Nat. Biomed. Eng., 6:94-104; herein incorporated by reference in its entirety)). As further discussed herein, the tumour environment or tumour microenvironment may have an elevated level of glucose compared to a healthy tissue or organ environment or microenvironment, leading to an elevated level of N-acetylglucosamine and / or lactate. Genetic mutations during tumour growth lead to the upregulation of lactate dehydrogenase A (LDH-A) that converts pyruvate to lactate. Lactate accumulation generates an acidic environment favouring tumour growth and suppressing immune cells activity (Cheung et al., Br J Cancer, 123, 261-267 (2020); Perez-Tomas & Perez-Guillen, Cancers, 2020, 12:3244; each herein incorporated by reference in its entirety). N-acetylglucosamine is a sugar moiety added to serine and / or threonine residues of cytosolic and nuclear proteins by O-GIcNAc transferase (OGT). N-acetylglucosamine is generated using both the pentose phosphate pathway (PPP) and the hexosamine biosynthetic pathway (HBP) fuelled by glucose and glutamine respectively (Ma & Vosseller, J Biol Chem., 2014; 289(50):34457-65; Le Minh etal., J. Biol. Chem., (2023), 299:105344; each herein incorporated by reference in its entirety). Accordingly, the tumour environment or tumour microenvironment may have an increased level of N-acetylglucosamine, N-acetylamine-6-phosphate, or lactate compared to a healthy tissue or organ environment or microenvironment (Gu et al., Cancer Res 1 August 2010; 70 (15): 6344-6351; Ma et al., J Biol Chem. 2014 Dec 12;289(50):34457-65; each herein incorporated by reference in its entirety). Ethanolamine is component of cellular membrane under the converted form of Phoshatidylethanolamine (PE). PE is the second most abundant phospholipid in mammalian cells. Free ethanolamine is found in body fluids and gastrointestinal tract after the degradation of PE-rich ingested food. During tumour development, free ethanolamine accumulates in the TME to contribute to tumour cells growth (Patel D, Witt SN. Oxid Med Cell Longev. 2017; 2017:4829180; Cheng et al., Targeting Phospholipid Metabolism in Cancer. Front Oncol. 2016; each herein incorporated by reference in its entirety). The tumour environment or tumour microenvironment may therefore have an elevated level of ethanolamine compared to a healthy tissue or organ environment or microenvironment (Osawa et al., Cell Reports, 2019, 29(l):89-103). Cyclic AMP (cAMP) may also be present in elevated levels in the tumour microenvironment or tumour environment compared to a healthy tissue or organ environment. To restrict expression of the extracellular matrix (ECM) degrading polypeptide by the cell to the tumour environment or tumour microenvironment, therefore, initiation of gene expression or transcription of a single nucleic acid transcript from the nucleic acid by the inducible promoter may suitably occur in hypoxic, anaerobic, anoxic, or microaerobic conditions and / or acidic conditions, and / or in the presence of N-acetylglucosamine, lactate, cyclic AMP (cAMP), and / or ethanolamine - in other words, the inducible promoter (such as a tumourinducible promoter) may suitably be a hypoxia-inducible promoter, a pH-inducible promoter, an N-acetylglucosamine-inducible promoter, a lactate-inducible promoter, cyclic AMP (cAMP), and / or an ethanolamine-inducible promoter. Initiation of gene expression or transcription of a single nucleic acid transcript from the nucleic acid by the inducible promoter may suitably not occur in oxic conditions and / or alkaline conditions, and / or in the absence of N-acetylglucosamine, lactate, cyclic AMP (cAMP), and / or ethanolamine.

[0096] As will be appreciated, a promoter comprises multiple promoter elements, each comprising discrete nucleotide sequences. Promoter elements may be selected from the group comprising or consisting of: an RNA polymerase binding site, a ribosome binding site (RBS), a -35 sequence, a -10 sequence, a Shine-Dalgarno sequence, a Pribnow Box, an operator sequence, a TATA box, a transcription factor binding site, a discriminator sequence, a -10 / -35 spacer sequence, or any combination thereof. Promoter elements may be arranged in a variety of different ways, and the sequences of each promoter element varied, in order to alter the activation specificity ( / .e., the conditions in which the promoter initiates or drives transcription of a single nucleic acid transcript from the nucleic acid downstream of the promoter) of a promoter or the strength ( / .e., the number of copiesof a single nucleic acid transcript transcribed under the control of the promoter in a given period of time) of a promoter. Promoter elements may also be arranged, and the sequences of each promoter element varied, in order to alter the translation initiation specificity or rate. Exemplary promoter elements that may be arranged or varied to alter the translation initiation or rate include, for example, the ribosome binding site (RBS), operator sequence(s), transcription factor binding site(s), discriminator sequence(s), -10 / -35 spacer sequence(s), or any combination thereof. Similarly, the length and / or GC of each promoter element may be varied to alter translation initiation, rate, efficiency, or tropism. For example, the length and / or GC content of a -10 / -35 spacer sequence or discriminator sequence may suitably be modified to achieve a desired translation initiation, rate, efficiency, or tropism.

[0097] For example, to design an inducible promoter that is a hypoxia-inducible promoter, a pH-inducible promoter, an N-acetylglucosamine-inducible promoter, a lactate-inducible promoter, and / or an ethanolamine-inducible promoter may be generated by modifying single nucleotides in the promoter sequence, for example in particular promoter elements, as disclosed in Davis et al., 2011, " Design, construction and characterization of a set of insulated bacterial promoters", Nucleic Acids Res., 39(3): 1131-1141, and Chen et al., 2018, " Tuning the dynamic range of bacterial promoters regulated by ligand-inducible transcription factors", Nat. Commun., 9:64 (each herein incorporated by reference in their entirety). Any promoter element may be modified; and any modification may be made. For example, a modification of a single nucleotide may include modification to introduce a single nucleotide polymorphism, a transition mutation, a transversion mutation, a base insertion, and / or a base deletion of the single nucleotide.

[0098] In some embodiments, the inducible promoter is a tumour-inducible promoter.

[0099] In some embodiments, the tumour-inducible promoter comprises an FNR binding motif. In some embodiments, the FNR binding motif comprises or consists a nucleotide sequence of SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244.

[0100] In some embodiments, the tumour-inducible promoter comprises an fnrS promoter or subsequence thereof. In some embodiments, the fnrS promoter or subsequence thereof comprises or consists of a nucleotide sequence of SEQ ID NO: 250 or SEQ ID NO: 257, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%,at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 50 or SEQ ID NO: 257.

[0101] Under anaerobic conditions, FNR (fumarate and nitrate reduction regulatory protein) dimerizes and binds to FNR binding motifs and / or the fnrS promoter, activating expression from promoters comprising these motifs. The presence of oxygen under normoxic conditions inhibits the ability of the FNR monomer to dimerize.

[0102] In some embodiments, the tumour-inducible promoter comprises a NagC binding motif. In some embodiments, the NagC binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 239, or SEQ ID NO: 259, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 239, or SEQ ID NO: 259.

[0103] NagC is a protein that controls the expression of enzymes involved in N-acetyl-glucosamine metabolism by binding to the NagC binding motif.

[0104] In some embodiments, the tumour-inducible promoter comprises a EutR binding motif. In some embodiments, the EutR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 261 or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 261.

[0105] EutR protein is expressed in an inactive form. EutR protein sense Ethanolamine by direct binding, which changes the conformation of EutR to the active form. By binding to eutR binding sites, the active EutR protein drives expression from promoters comprising the eutR binding site.

[0106] In some embodiments, the tumour-inducible promoter comprises an lldR binding motif. In some embodiments, the lldR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 248, SEQ ID NO: 249, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249.

[0107] In the absence of lactate, the LldR protein dimerises and binds to IldR binding motifs in DNA. This represses expression of promoters comprising the IldR binding motif. In thepresence of lactate, the LldR protein changes the conformation and dissociates from the IldR binding motif, inducing promoter activation.

[0108] In some embodiments, the tumour-inducible promoter comprises a CRP binding motif. In some embodiments, the CRP binding motif comprises or consists of a nucleotide sequence encoded by: SEQ ID NO: 245 or SEQ ID NO: 246; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 245 or SEQ ID NO: 246.

[0109] In some embodiments, the tumour-inducible promoter comprises a grcA promoter or fragment thereof. In some embodiments, the grcA promoter or fragment thereof comprises or consists of a nucleotide sequence of: SEQ ID NO: 253 or SEQ ID NO: 254, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249.

[0110] In some embodi ments, the tum< Dur-inducible pror noter comprises or consists of a nucleotide sequer ice of: SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO; 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO; 135, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO; 145, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO; 155, SEQ ID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO; 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO; 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO: 179, SEQ ID NO: 180, SEQ ID NO: 181, SEQ ID NO: 182, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO; 185, SEQ ID NO: 186, SEQ ID NO: 187, SEQ ID NO: 188, SEQ ID NO: 189, SEQ ID NO: 190, SEQ ID NO: 191, SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO; 195, SEQ ID NO: 196, SEQ ID NO: 197, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO; 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210, SEQ ID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, or SEQ ID NO: 278; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least99% identical to SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO; 125, SEQ ID NO 126, SEQ ID NO 127, SEQ ID NO: 128, SEQ ID NO 129, SEQ ID NO: 130, SEQ ID NO 131, SEQ ID NO 132, SEQ ID NO: 133, SEQ ID NO; 135, SEQ ID NO: 136, SEQ ID NO 139, SEQ ID NO 140, SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO; 145, SEQ ID NO 146, SEQ ID NO 147, SEQ ID NO: 148, SEQ ID NO 149, SEQ ID NO: 150, SEQ ID NO 151, SEQ ID NO 152, SEQ ID NO: 153, SEQ ID NO 154, SEQ ID NO; 155, SEQ ID NO 156, SEQ ID NO 157, SEQ ID NO: 158, SEQ ID NO 159, SEQ ID NO: 160, SEQ ID NO 161, SEQ ID NO 162, SEQ ID NO: 163, SEQ ID NO 164, SEQ ID NO; 165, SEQ ID NO 166, SEQ ID NO 167, SEQ ID NO: 168, SEQ ID NO 169, SEQ ID NO: 170, SEQ ID NO 171, SEQ ID NO 172, SEQ ID NO: 173, SEQ ID NO 174, SEQ ID NO; 175, SEQ ID NO 176, SEQ ID NO 177, SEQ ID NO: 178, SEQ ID NO 179, SEQ ID NO: 180, SEQ ID NO 181, SEQ ID NO 182, SEQ ID NO: 183, SEQ ID NO 184, SEQ ID NO; 185, SEQ ID NO 186, SEQ ID NO 187, SEQ ID NO: 188, SEQ ID NO 189, SEQ ID NO: 190, SEQ ID NO 191, SEQ ID NO 192, SEQ ID NO: 193, SEQ ID NO 194, SEQ ID NO; 195, SEQ ID NO 196, SEQ ID NO 197, SEQ ID NO: 198, SEQ ID NO 199, SEQ ID NO: 200, SEQ ID NO 201, SEQ ID NO 202, SEQ ID NO: 203, SEQ ID NO 204, SEQ ID NO; 205, SEQ ID NO 206, SEQ ID NO 207, SEQ ID NO: 208, SEQ ID NO 209, SEQ ID NO: 210, SEQ ID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, or SEQ ID NO: 278;.

[0111] In some embodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 160, SEQ ID NO: 182, SEQ ID NO: 190, SEQ ID NO: 128, SEQ ID NO: 125, SEQ ID NO: 131, SEQ ID NOL 132, SEQ ID NO: 133, SEQ ID NO: 144, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 185: SEQ ID NO: 204, SEQ ID NO: 209, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 160, SEQ ID NO: 182, SEQ ID NO: 190, SEQ ID NO: 128, SEQ ID NO: 125, SEQ ID NO: 131, SEQ ID NOL 132, SEQ ID NO: 133, SEQ ID NO: 144, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 185: SEQ ID NO: 204, SEQ ID NO: 209.

[0112] In some embodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 160, SEQ ID NO: 182, SEQ ID NO: 190, SEQ ID NO: 128, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 160, SEQ ID NO: 182, SEQ ID NO: 190, SEQ ID NO: 128.

[0113] In some embodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 160. In some embodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 182. In someembodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 190. In some embodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 128. In some embodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 125. In some embodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 131. In some embodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NOL 132. In some embodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 133. In some embodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 144. In some embodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 170. In some embodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 171. In some embodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 185: SEQ ID NO: 204. In some embodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 209,

[0114] In some embodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278.

[0115] In some embodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 275. In some embodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 276. In some embodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 277. In some embodiments, the tumour-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 278.

[0116] In some embodiments, the tumour-inducible promoter is a hypoxia-inducible promoter. In some embodiments, the hypoxia-inducible promoter comprises an FNR binding motif. In some embodiments, the FNR binding motif comprises a or consists of a nucleotide sequence of SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244.In some embodiments, the hypoxia-inducible promoter comprises an fnrS promoter or subsequence thereof. In some embodiments, the fnrS promoter or subsequence thereof comprises or consists of a nucleotide sequence of SEQ ID NO: 250 or SEQ ID NO: 257, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 250 or SEQ ID NO: 257.

[0117] In some embodiments, the hypoxia-inducible promoter comprises a grcA promoter or fragment thereof comprises or consists of a nucleotide sequence of: SEQ ID NO: 253 or SEQ ID NO: 254, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249.

[0118] In some embodiments, the hypoxia-inducible promoter is a hypoxia-inducible promoter disclosed in International Patent Application No. PCT / GB2024 / 051897. Accordingly, in some embodiments, optionally the hypoxia-inducible promoter is selected from the group comprising or consisting of: LOR9, LOR7, LOR1, pflE, pepT, YbiY, and pvhb.

[0119] In some embodiments, the hypoxia-inducible promoter is activated and drives expression of the heterologous ECM degrading polypeptide at an oxygen concentration of:

[0120] a) between about 0% and about 2%, about 0.2% and about 1.8%, about 0.4% and about 1.6%, about 0.6% and about 1.4%, about 0.8% and about 1.2%, about 0% and about 1%, about 0.2% and about 0.8%, about 0.4 and about 0.6%, 1% and 2%, about 1.2% and about 1.8%, about 1.4% and about 1.6%;

[0121] b) between 0% and 2%, 0.2% and 1.8%, 0.4% and 1.6%, 0.6% and 1.4%, 0.8% and 1.2%, 0% and 1%, 0.2% and 0.8%, 0.4 and 0.6%, 1% and 2%, 1.2% and 1.8%, 1.4% and 1.6%;

[0122] c) below about 2%, below about 1.5%, below about 1.6%, below about 1.4%, below about 1.2%, below about 1%, below about 0.8%, below about 0.6%, below about 0.4%, below about 0.2% or lower;

[0123] d) below 2%, below 1.5%, below 1.6%, below 1.4%, below 1.2%, below 1%, below 0.8%, below 0.6%, below 0.4%, below 0.2% or lower;

[0124] e) about 2%, about 1.8%, about 1.6%, about 1.4%, about 1.2%, about 1%, about 0.8%, about 0.6%, about 0.4%, about 0.2%; and / or

[0125] f) 2%, 1.8%, 1.6%, 1.4%, 1.2%, 1%, 0.8%, 0.6%, 0.4%, 0.2%.

[0126] In some embodiments, the hypoxia-inducible promoter is not activated and does not drive expression of the heterologous ECM degrading polypeptide in normoxia; optionallywherein the hypoxia-inducible promoter is not activated and does not drive expression of the heterologous ECM degrading polypeptide at an oxygen concentration of:

[0127] a) above about 6%, above about 5%, above about 4%, or above about 3%;

[0128] b) above 6%, above 5%, above 4%, or above 3%;

[0129] c) about 6%, about 5%, about 4%, about 3%; and / or

[0130] d) 6%, 5%, 4%, 3%.

[0131] Hypoxic conditions may be simulated in vitro by culturing a cell in liquid culture media, where the surface of the air-liquid interface is completely covered or substantially covered with a layer of mineral oil. In some embodiments, the hypoxia-inducible promoter is activated and drives expression of the heterologous ECM degrading polypeptide when the recombinant cell is incubated in culture media under mineral oil. Appropriate methods for culturing a cell in such conditions may be determined by the person skilled in the art; however, in some embodiments the cell is optionally incubated at 37°C for 18 hours.

[0132] In some embodiments the hypoxia-inducible promoter comprises a ribosome binding site (RBS). In some embodiments, the ribosome binding site (RBS) comprises or consists a nucleotide sequence of: SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, or SEQ ID NO: 105; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, or SEQ ID NO: 105.

[0133] In some embodiments, the hypoxia-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12,SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ D NO: 132, SEQ UD NO: 133, SEQ ID NO 135, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO; 155, SEQ ID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO; 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO; 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO: 179, SEQ ID NO: 180, SEQ ID NO: 189, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO; 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210, SEQ ID NO: 275, or SEQ ID NO: 278; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 54, SEQ ID NO: 55, or SEQ ID NO: 56, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ D NO: 132, SEQ UD NO: 133, SEQ ID NO 135, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: 150, SEQ ID NO: 179, SEQ ID NO: 180, SEQ ID NO: 189, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO; 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210, SEQ ID NO: 275, or SEQ ID NO: 278.

[0134] In some embodiments, the hypoxia-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ D NO: 132, SEQ UD NO: 133, SEQ ID NO 135, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: SEQ ID NO: 150, SEQ ID NO: 179, SEQ ID NO: 180, SEQ ID NO: 275, or SEQ ID NO: 278; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56,SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ D NO: 132, SEQ UD NO: 133, SEQ ID NO 135, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: SEQ ID NO: 150, SEQ ID NO: 179, SEQ ID NO: 180, SEQ ID NO: 275, or SEQ ID NO: 278.

[0135] In some embodiments, the hypoxia-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 128, or SEQ ID NO: 278, SEQ ID NO: 125, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO: 144, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 204, or SEQ ID NO: 209; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 128, or SEQ ID NO: 278, SEQ ID NO: 125, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO: 144, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 204, or SEQ ID NO: 209.

[0136] In some embodiments, the hypoxia-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 128, or SEQ ID NO: 278; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 128, or SEQ ID NO: 278.

[0137] In some embodiments, the hypoxia-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 160. In some embodiments, the hypoxia-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 275. In some embodiments, the hypoxia-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 128. In some embodiments, the hypoxia-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 278. In some embodiments, the hypoxia-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 125. In some embodiments, the hypoxia-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 131. In some embodiments, the hypoxia-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 132. In some embodiments, the hypoxia-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 133. In some embodiments, the hypoxia-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 144. In some embodiments, the hypoxia-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 170. In some embodiments, the hypoxia-inducible promoter comprises or consists ofa nucleotide sequence of SEQ ID NO: 171. In some embodiments, the hypoxia-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 204. In some embodiments, the hypoxia-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 209.

[0138] In some embodiments, the tumour-inducible promoter is an N-acetylglucosamine-inducible promoter. In some embodiments, the N-acetylglucosamine-inducible promoter comprises a NagC binding motif. In some embodiments, the NagC binding motif comprises or consists of a nucleotide sequence of: SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 239, or SEQ ID NO: 259; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 239, or SEQ ID NO: 259.

[0139] In some embodiments, the N-acetylglucosamine-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 169, SEQ ID NO: 172, SEQ ID NI: 173, SEQ ID NO: 194, SEQ ID NO: 196, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, or SEQ ID NO: 207; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 169, SEQ ID NO: 172, SEQ ID NI: 173, SEQ ID NO: 194, SEQ ID NO: 196, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, or SEQ ID NO: 207.

[0140] In some embodiments, the N-acetylglucosamine-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 144 or SEQ ID NO: 204; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 144 or SEQ ID NO: 204.

[0141] In some embodiments, the N-acetylglucosamine-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 144. In some embodiments, the N-acetylglucosamine-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 204.

[0142] In some embodiments, the N-acetylglucosamine-inducible promoter is activated and drives expression of the extracellular matrix (ECM) degrading polypeptide in the presence of acompound selected from the group comprising or consisting of: N-acetylglucosamine, N-acetylglucosamine-6-phosphate. In some embodiments, the N-acetylglucosamine-inducible promoter is activated and drives expression of the extracellular matrix (ECM) degrading polypeptide in the presence of N-acetylglucosamine. In some embodiments, the N-acetylglucosamine-inducible promoter is activated and drives expression of the extracellular matrix (ECM) degrading polypeptide at an N-acetylglucosamine concentration of:

[0143] a) at least 0.01 mM, at least 0.02 mM, at least 0.03 mM, at least 0.04 mM, at least 0.05 mM, at least 0.06 mM, at least 0.07 mM, at least 0.08 mM, at least 0.09 mM, at least 0.1 mM, at least 0.2 mM, at least 0.3 mM, at least 0.4 mM, at least 0.5 mM, at least 0.6 mM, at least 0.7 mM, at least 0.8 mM, at least 0.9 mM, at least 1 mM, at least 5 mM, at least 6 mM, at least 7 mM, at least 8 mM, at least 9 mM, at least 10 mM, or more; b) about 0.01 mM, about 0.02 mM, about 0.03 mM, about 0.04 mM, about 0.05 mM, about 0.06 mM, about 0.07 mM, about 0.08 mM, about 0.09 mM, about 0.1 mM, about 0.2 mM, about 0.3 mM, about 0.4 mM, about 0.5 mM, about 0.6 mM, about 0.7 mM, about 0.8 mM, about 0.9 mM, about 1 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, or about 10 mM;

[0144] c) 0.01 mM, 0.02 mM, 0.03 mM, 0.04 mM, 0.05 mM, 0.06 mM, 0.07 mM, 0.08 mM, 0.09 mM, 0.1 mM, 0.2 mM, 0.3 mM, 0.4 mM, 0.5 mM, 0.6 mM, 0.7 mM, 0.8 mM, 0.9 mM, 1 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, or 10 mM.

[0145] In some embodiments, the tumour-inducible promoter is an ethanolamine-inducible promoter, the ethanolamine-inducible promoter comprises a EutR binding motif. In some embodiments, the EutR binding motif comprises or consists of a nucleotide sequence of: SEQ ID NO: 261 or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 261.

[0146] In some embodiments, the ethanolamine-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, or SEQ ID NO: 197; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, or SEQ ID NO: 197.

[0147] In some embodiments, the ethanolamine-inducible promoter comprises or consists of a nucleotide a sequence of: SEQ ID NO: 192 or SEQ ID NO: 197; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 192 or SEQ ID NO: 197.

[0148] In some embodiments, the ethanolamine-inducible promoter is activated and drives expression of the extracellular matrix (ECM) degrading polypeptide in the presence of a compound selected from the group comprising or consisting of: ethanolamine, adenosylcobalamin, and glucose. In some embodiments, the ethanolamine-inducible promoter is activated and drives expression of the extracellular matrix (ECM) degrading polypeptide in the presence of ethanolamine.

[0149] In some embodiments, In some embodiments, the ethanolamine-inducible promoter is activated and drives expression of the extracellular matrix (ECM) degrading polypeptide at an ethanolamine concentration of:

[0150] a) at least 0.1 mM, at least 0.2 mM, at least 0.3 mM, at least 0.4 mM, at least 0.5 mM, at least 0.6 mM, at least 0.7 mM, at least 0.8 mM, at least 0.9 mM, at least 1 mM, at least 5 mM, at least 6 mM, at least 7 mM, at least 8 mM, at least 9 mM, at least 10 mM, at least 20 mM, at least 30 mM, at least 40 mM, at least 50 mM, at least 60 mM, at least 70 mM, at least 80 mM, at least 90 mM, at least 100 mM, or more;

[0151] b) about 0.1 mM, about 0.2 mM, about 0.3 mM, about 0.4 mM, about 0.5 mM, about 0.6 mM, about 0.7 mM, about 0.8 mM, about 0.9 mM, about 1 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, about 10 mM, about 20 mM, about 30 mM, about 40 mM, about 50 mM, about 60 mM, about 70 mM, about 80 mM, about 90 mM, or about 100 mM;

[0152] c) 0.1 mM, 0.2 mM, 0.3 mM, 0.4 mM, 0.5 mM, 0.6 mM, 0.7 mM, 0.8 mM, 0.9 mM, 1 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, or 100 mM.

[0153] In some embodiments, the tumour-inducible promoter is a lactate-inducible promoter. In some embodiments, the lactate-inducible promoter comprises an IldR binding motif. In some embodiments, the IldR binding motif comprises or consists of a nucleotide sequence of: SEQ ID NO: 248, SEQ ID NO: 249, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249.

[0154] In some embodiments, the lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO: 155, SEQ ID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQID NO; 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO; 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO: 189, SEQ ID NO: 190, SEQ ID NO: 191, SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 201, SEQ ID NO: 275, or SEQ ID NO: 277; or a sequence is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO: 155, SEQ ID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO; 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO; 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO: 189, SEQ ID NO: 190, SEQ ID NO: 191, SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 201, SEQ ID NO: 275, or SEQ ID NO: 277.

[0155] In some embodiments, the lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 190, SEQ ID NO: 277, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 204, or SEQ ID NO: 209; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 190, SEQ ID NO: 277, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 204, or SEQ ID NO: 209.

[0156] In some embodiments, the lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 190, or SEQ ID NO: 277; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 190, or SEQ ID NO: 277.

[0157] In some embodiments, the lactate-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 160. In some embodiments, the lactate-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 275. In some embodiments,the lactate-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 190. In some embodiments, the lactate-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 277. In some embodiments, the lactate-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 170. In some embodiments, the lactate-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 171. In some embodiments, the lactate-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 204. In some embodiments, the lactate-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 209.

[0158] In some embodiments, the lactate-inducible promoter is activated and drives expression of the extracellular matrix (ECM) degrading polypeptide in the presence of lactate. In some embodiments, the lactate-inducible promoter is activated and drives expression of the extracellular matrix degrading enzyme at a lactate concentration disclosed in Saab et al., 2023, eLife, 12:e81289, Sullivan etal., 2019, eLife, 8:e44235, and / or Cheung etal., 2020, Br. J. Cancer, 123:261-267 (each incorporated herein by reference in their entirety). In some embodiments, the lactate-inducible promoter is activated and drives expression of the extracellular matrix (ECM) degrading polypeptide at a lactate concentration of: a) at least ImM, at least 2mM, at least 3 mM, at least 4 mM, at least 5 mM, at least 6 mM, at least 7 mM, at least 8 mM, at least 9 mM, at least 10 mM, at least 20 mM, at least 30 mM, at least 40 mM, at least 50 mM, at least 60 mM, at least 70 mM, at least 80 mM, at least 90 mM, at least 100 mM, at least 200 mM, at least 300 mM, at least 400 mM, at least 500 mM, at least 600 mM, at least 700 mM, at least 800 mM, at least 900 mM, at least 1 M, or higher;

[0159] b) about ImM, about 2mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, about 10 mM, about 20 mM, about 30 mM, about 40 mM, about 50 mM, about 60 mM, about 70 mM, about 80 mM, about 90 mM, about 100 mM, about 200 mM, about 300 mM, about 400 mM, about 500 mM, about 600 mM, about 700 mM, about 800 mM, about 900 mM, or about 1 M; and / or

[0160] c) 1 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 200 mM, 300 mM, 400 mM, 500 mM, 600 mM, 700 mM, 800 mM, 900 mM, or 1 M.

[0161] In some embodiments, the tumour-inducible promoter is a pH-inducible promoter.

[0162] In some embodiments, the pH inducible promoter is selected from the group comprising or consisting of: LPR7, LPR1, LPR9, Stm1787, hyaA, P170-IL, P170-MG, Pl, P2, and P3.In some embodiments, the pH-inducible promoter is activated and drives expression of the heterologous ECM degrading polypeptide at an acidic pH; optionally

[0163] wherein the acidic pH is:

[0164] a) between about pH 5.5 and about pH 7, about pH 5.6 and about pH 6.9, about pH 5.7 and about pH 6.8, about pH 5.8 and about pH 6.7, about pH 5.9 and about pH 6.6, about pH 6.0 and about pH 6.5, about pH 6.1 and about pH 6.4, about pH 6.2 and about pH 6.3;

[0165] b) between pH 5.5 and pH 7, pH 5.6 and pH 6.9, pH 5.7 and pH 6.8, pH 5.8 and pH 6.7, pH 5.9 and pH 6.6, pH 6.0 and pH 6.5, pH 6.1 and pH 6.4, pH 6.2 and pH 6.3;

[0166] c) below about pH 7, below about pH 6.9, below about pH 6.8, below about pH 6.7, below about pH 6.6, below about pH 6.5, below about pH 6.4, below about pH 6.3, below about pH 6.2, below about pH 6.1, below about pH 6.0, below about pH 5.9, below about pH 5.8, below about pH 5.7, below about pH 5.6, or lower;

[0167] d) below pH 7, below pH 6.9, below pH 6.8, below pH 6.7, below pH 6.6, below pH 6.5, below pH 6.4, below pH 6.3, below pH 6.2, below pH 6.1, below pH 6.0, below pH 5.9, below pH 5.8, below pH 5.7, below pH 5.6;

[0168] e) about pH 5.5, about pH 5.6, about pH 5.7, about pH 5.8, about pH 5.9, about pH 6.0, about pH 6.1, about pH 6.2, about pH 6.3, about pH6.4, about pH 6.5, about pH 6.6, about pH 6.7, about pH 6.8, about pH 6.9, or about pH 7; and / or

[0169] f) pH 5.5, pH 5.6, pH 5.7, pH 5.8, pH 5.9, pH 6.0, pH 6.1, pH 6.2, pH 6.3, pH6.4, pH 6.5, pH 6.6, pH 6.7, pH 6.8, pH 6.9, or pH 7.

[0170] In some embodiments, the pH-inducible promoter is activated and drives expression of the heterologous ECM degrading polypeptide at an acidic pH; optionally

[0171] wherein the acidic pH is:

[0172] a) between about pH 5.9 and about pH 6.5, about pH 5.95 and about pH 6.45, about pH 6.0 and about pH 6.4, about pH 6.05 and about pH 6.35, about pH 6.1 and about 6.3, or about pH 6.15 and about pH 6.25;

[0173] b) between pH 5.9 and pH 6.5, pH 5.95 and pH 6.45, pH 6.0 and pH 6.4, pH 6.05 and pH 6.35, pH 6.1 and 6.3, or pH 6.15 and pH 6.25;

[0174] c) about pH 5.9, about pH 5.95, about pH 6.0, about pH 6.05, about pH 6.1, about pH 6.15, about pH 6.2, about pH 6.25, about pH 6.3, about pH 6.35, about pH 6.4, about pH 6.45, or about pH 6.5; and / or

[0175] d) pH 5.9, pH 5.95, pH 6.0, pH 6.05, pH 6.1, pH 6.15, pH 6.2, pH 6.25, pH 6.3, pH 6.35, pH 6.4, pH 6.45, or pH 6.5.

[0176] In some embodiments, the pH-inducible promoter comprises a ribosome binding site (RBS). In some embodiments, the ribosome binding site (RBS) comprises or consists of anucleotide sequence of SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, or SEQ ID NO: 105; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, or SEQ ID NO: 105.

[0177] In some embodiments, the pH-inducible promoter is comprises or consists of a nucleotide sequence of: SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO 181, SEQ ID NO: 182, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 187, SEQ ID NO: 188, SEQ ID NO: 190, SEQ ID NO: 191, SEQ ID NO: 276, or SEQ ID NO: 277; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, or SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO 181, SEQ ID NO: 182, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 187, SEQ ID NO: 188, SEQ ID NO: 190, SEQ ID NO: 191, SEQ ID NO: 276, or SEQ ID NO: 277.

[0178] In some embodiments, the pH-inducible promoter comprises or consists of nucleotide sequence of: SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO 181, SEQ ID NO: 182, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO: 185, SEQ ID NO: 186, or SEQ ID NO: 187, SEQ ID NO: 276, or SEQ ID NO: 277; or a nucleotide sequence that is at least50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO 181, SEQ ID NO: 182, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO: 185, SEQ ID NO: 186, or SEQ ID NO: 187, SEQ ID NO: 276, or SEQ ID NO: 277.

[0179] In some embodiments, the pH-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 182, SEQ ID NO: 276, SEQ ID NO: 190, SEQ ID NO: 277, or SEQ ID NO: 185; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 182, SEQ ID NO: 276, SEQ ID NO: 190, SEQ ID NO: 277, or SEQ ID NO: 185.

[0180] In some embodiments, the pH-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 182, SEQ ID NO: 276, SEQ ID NO: 190, or SEQ ID NO: 277; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 182, SEQ ID NO: 276, SEQ ID NO: 190, or SEQ ID NO: 277.

[0181] In some embodiments, the pH-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 182. In some embodiments, the pH-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 276. In some embodiments, the pH-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 190. In some embodiments, the pH-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 277. In some embodiments, the pH-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 185.

[0182] In some embodiments, the pH-inducible promoter is not activated and does not drive expression of the heterologous ECM degrading polypeptide at an alkaline pH. The term "alkaline pH" is known to the person skilled in the art. A solution has an "alkaline pH" if it has a pH of between pH 7 and pH 14.

[0183] In some embodiments, the tumour-inducible promoter is a cAMP-inducible promoter. In some embodiments, the tumour-inducible promoter comprises a CRP binding motif. In some embodiments, the CRP binding motif comprises or consists of a nucleotide sequence of: SEQ ID NO: 245 or SEQ ID NO: 246; or a nucleotide sequence that is at least 50%, atleast 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 245 or SEQ ID NO: 246.

[0184] In some embodiments, the cAMP-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 170, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 189, SEQ ID NO: 208, SEQ ID NO: 209, or SEQ ID NO: 210; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to: SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 170, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 189, SEQ ID NO: 208, SEQ ID NO: 209, or SEQ ID NO: 210.

[0185] In some embodiments, the cAMP-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 170 or SEQ ID NO: 209; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to: SEQ ID NO: 170 or SEQ ID NO: 209.

[0186] In some embodiments, the cAMP-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 170. In some embodiments, cAMP-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 209.

[0187] In some embodiments, the cAMP-inducible promoter is activated and drives expression of the extracellular matrix (ECM) degrading polypeptide in the presence of a compound selected from the group comprising or consisting of: cAMP. In some embodiments, the lactate-inducible promoter is activated and drives expression of the extracellular matrix (ECM) degrading polypeptide in the presence of cAMP.

[0188] Advantageously, the expression of the extracellular matrix (ECM) degrading enzyme is controlled by multiple conditions present in the tumour microenvironment.

[0189] In some embodiments, the tumour-inducible promoter is a hypoxia- and N-acetylglucosamine-inducible promoter. In some embodiments, the hypoxia- and N-acetylglucosamine-inducible promoter comprises an FNR binding motif and / or an fnrS promoter or subsequence thereof and / or a grcA promoter or fragment thereof, and a NagC binding motif. In some embodiments:i) a) the FNR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244;

[0190] b) the fnrS promoter or subsequence thereof comprises or consists of a nucleotide sequence of SEQ ID NO: 250 or SEQ ID NO: 257, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 250 or SEQ ID NO: 257; and / or

[0191] c) the grcA promoter or fragment thereof comprises or consists of a nucleotide sequence of: SEQ ID NO: 253 or SEQ ID NO: 254, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249.

[0192] and / or

[0193] ii) the NagC binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 239, or SEQ ID NO: 259, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 239, or SEQ ID NO: 259.

[0194] In some embodiments, the hypoxia- and N-acetylglucosamine-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 169, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 196, SEQ ID NO: 198, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 169, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 196, SEQ ID NO: 198, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207.In some embodiments, the hypoxia- and N-acetylglucosamine-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 198, SEQ ID NO: 200, or SEQ ID NO: 204; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 198, SEQ ID NO: 200, or SEQ ID NO: 204.

[0195] In some embodiments, the hypoxia- and N-acetylglucosamine-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 144 or SEQ ID NO: 204; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 144 or SEQ ID NO: 204.

[0196] In some embodiments, the hypoxia- and N-acetylglucosamine-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 144. In some embodiments, the hypoxia- and N-acetylglucosamine-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 204.

[0197] The hypoxia- and N-acetylglucosamine-inducible promoter may be activated and drive expression of the extracellular matrix degrading (ECM) degrading enzyme in the presence of both hypoxic conditions as described herein and N-acetlyglucosamine or N-acetylfglucosamine-6-phosphate.

[0198] In some embodiments, the tumour-inducible promoter is a hypoxia- and ethanolamine-inducible promoter. In some embodiments, the hypoxia- and ethanolamine-inducible promoter comprises an FNR binding motif and / or an fnrS promoter or subsequence thereof and / or a grcA promoter or fragment thereof, and a EutR binding motif. In some embodiments:

[0199] i) a) the FNR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244;

[0200] b) the fnrS promoter or subsequence thereof comprises or consists of a nucleotide sequence of SEQ ID NO: 250 or SEQ ID NO: 257, or a sequence that isat least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 250 or SEQ ID NO: 257; and / or

[0201] c) the grcA promoter or fragment thereof comprises or consists of a nucleotide sequence of: SEQ ID NO: 253 or SEQ ID NO: 254, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249

[0202] and / or

[0203] ii) the EutR binding motif comprises a sequence of SEQ ID NO: 261 or or consists of a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 261.

[0204] In some embodiments, the hypoxia- and ethanolamine-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 195 or SEQ ID NO: 196; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 195 or SEQ ID NO: 196.

[0205] The hypoxia- and ethanolamine-inducible promoter may be activated and drive expression of the extracellular matrix degrading (ECM) degrading enzyme in the presence of both hypoxic conditions as described herein and ethanolamine.

[0206] In some embodiments, the tumour-inducible promoter is a hypoxia- and lactate-inducible promoter. In some embodiments, the hypoxia and lactate-inducible promoter comprises an FNR binding motif and / or an fnrS promoter or subsequence thereof and / or a grcA promoter or fragment thereof, and an IldR binding motif. In some embodiments:

[0207] i) a) the FNR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244;

[0208] b) the fnrS promoter or subsequence thereof comprises or consists of a nucleotide sequence of SEQ ID NO: 250 or SEQ ID NO: 257, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%,at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 250 or SEQ ID NO: 257; and / or

[0209] c) the grcA promoter or fragment thereof comprises or consists of a nucleotide sequence of: SEQ ID NO: 253 or SEQ ID NO: 254, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249;

[0210] and / or

[0211] ii) the lldR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 248, SEQ ID NO: 249, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249.

[0212] In some embodiments, the hypoxia- and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 156, SEQ ID NO: 157, SEQ IDNO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO: 189, SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206 SEQ ID NO: 207, or SEQ ID NO: 275; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 156, SEQ ID NO: 157, SEQ IDNO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO: 189, SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206. Or SEQ ID NO: 207, or SEQ ID NO: 275.

[0213] In some embodiments, the hypoxia- and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 156, SEQ ID NO: 157, SEQ IDNO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 171, SEQ ID NO: 174, SEQ ID NO: 175, SEQ IDNO: 178, or SEQ ID NO: 275; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 156, SEQ ID NO: 157, SEQ IDNO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 171, SEQ ID NO: 174, SEQ ID NO: 175, or SEQ ID NO: 178, or SEQ ID NO: 275.

[0214] In some embodiments, the hypoxia- and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 204, or SEQ ID NO: 209; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 160 SEQ ID NO: 275, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 204, or SEQ ID NO: 209.

[0215] In some embodiments, the hypoxia- and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 160, or SEQ ID NO: 275; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 160 or SEQ ID NO: 275.

[0216] In some embodiments, the hypoxia- and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 160. In some embodiments, the hypoxia- and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 275. In some embodiments, the hypoxia- and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 170. In some embodiments, the hypoxia-and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 171. In some embodiments, the hypoxia- and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 204. In some embodiments, the hypoxia- and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 209.

[0217] The hypoxia- and lactate-inducible promoter may be activated and drive expression of the extracellular matrix degrading (ECM) degrading enzyme in the presence of both hypoxic conditions as described herein and lactate.

[0218] In some embodiments, the inducible promoter (such as a tumour-inducible promoter) is a hypoxia- and pH-inducible promoter. In some embodiments, the hypoxia- and pH-inducible promoter comprises an FNR binding motif and / or an fnrS promoter or subsequence thereof and / or a grcA promoter or fragment thereof. In some embodiments: i) a) the FNR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244;

[0219] b) the fnrS promoter or subsequence thereof comprises or consists of a nucleotide sequence of SEQ ID NO: 250 or SEQ ID NO: 257, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 250 or SEQ ID NO: 257; and / or

[0220] c) the grcA promoter or fragment thereof comprises or consists of a nucleotide sequence of SEQ ID NO: 253 or SEQ ID NO: 254, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249.

[0221] In some embodiments, the pH-inducible and hypoxia-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 188; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 188.

[0222] In some embodiments, the pH-inducible and hypoxia-inducible promoter is a pH-inducible and hypoxia-inducible promoter disclosed in International Patent Application No. PCT / GB2024 / 051897. Accordingly, in some embodiments, the pH-inducible and hypoxiainducible promoter is LOR9. In some embodiments:

[0223] a) the LOR9 promoter comprises a ribosome binding site (RBS) encoded by a nucleotide sequence of SEQ ID NO: 73, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 73; and / or b) the LOR9 promoter is encoded by a nucleotide sequence of SEQ ID NO: 55 or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 55.The pH-inducible promoter and hypoxia-inducible promoter may be activated at the same pH and O2concentration as the pH-inducible promoters and hypoxia-inducible promoters provided herein.

[0224] In some embodiments, the tumour-inducible promoter is a hypoxia- and cAMP-inducible promoter. In some embodiments, the hypoxia- and cAMP-inducible promoter comprises an FNR binding motif and / or an fnrS promoter or subsequence thereof and / or grcA promoter or fragment thereof, and a CRP binding motif. In some embodiments:

[0225] i) a) the FNR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244;

[0226] b) the fnrS promoter or subsequence thereof comprises or consists of a nucleotide sequence of SEQ ID NO: 250 or SEQ ID NO: 257, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 250 or SEQ ID NO: 257; and / or

[0227] c) the grcA promoter or fragment thereof comprises or consists of a nucleotide sequence of SEQ ID NO: 253 or SEQ ID NO: 254, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249;

[0228] and / or

[0229] ii) the CRP binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 245 or SEQ ID NO: 246; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 245 or SEQ ID NO: 246.

[0230] In some embodiments, the hypoxia- and cAMP-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 170, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 189, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 170, SEQID NO: 176, SEQ ID NO: 177, SEQ ID NO: 189, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210.

[0231] In some embodiments, the hypoxia- and cAMP-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 148, SEQ ID NO: 149; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 148, or SEQ ID NO: 149.

[0232] In some embodiments, the hypoxia- and cAMP-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 170, or SEQ ID NO: 209; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 170, or SEQ ID NO: 209.

[0233] In some embodiments, the hypoxia- and cAMP-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 148. the hypoxia- and cAMP-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 149. the hypoxia- and cAMP-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 170. the hypoxia- and cAMP-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 209.

[0234] The hypoxia- and cAMP-inducible promoter may be activated and drive expression of the extracellular matrix degrading (ECM) degrading enzyme in the presence of both hypoxic conditions as described herein and cAMP.

[0235] In some embodiments, the tumour-inducible promoter is a pH- and lactate-inducible promoter. In some embodiments, the pH- and lactate-inducible promoter comprises an IldR binding motif. In some embodiments:

[0236] i) the IldR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 248, SEQ ID NO: 249, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249.

[0237] In some embodiments, the pH- and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 190, SEQ ID NO: 191, or SEQ ID NO: 277; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, atleast 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 190, SEQ ID NO: 191, or SEQ ID NO: 277.

[0238] In some embodiments, the pH- and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 190 or SEQ ID NO: 277; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 190 or SEQ ID NO: 277.

[0239] In some embodiments, the pH- and lactate-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 190. In some embodiments, the pH- and lactate-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO: 277.

[0240] The pH- and lactate-inducible promoter may be activated and drive expression of the extracellular matrix degrading (ECM) degrading enzyme in the presence of both acidic pH as described herein and lactate.

[0241] In some embodiments, the tumour-inducible promoter is a hypoxia-, N-acetylglucosamine, and lactate-inducible promoter. In some embodiments, the hypoxia-, N-acetylglucosamine, and lactate-inducible promoter comprises an FNR binding motif and / or an fnrS promoter or subsequence thereof and / or a grcA promoter or fragment thereof, a NagC binding motif, and an IldR binding motif. In some embodiments:

[0242] i) a) the FNR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244; and / or

[0243] b) the fnrS promoter or subsequence thereof comprises or consists of a nucleotide sequence of SEQ ID NO: 250 or SEQ ID NO: 257, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 250 or SEQ ID NO: 257;

[0244] c) the grcA promoter or subsequence thereof comprises or consists of a nucleotide sequence of SEQ ID NO: 253 or SEQ ID NO: 254, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%,at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249;

[0245] and / or

[0246] ii) the NagC binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 239, or SEQ ID NO: 259, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 239, or SEQ ID NO: 259; and / or

[0247] iii) the lldR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 248, SEQ ID NO: 249, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249.

[0248] In some embodiments, the tumour-inducible promoter is a hypoxia-, N-acetylglucosamine, and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 169, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 205, SEQ ID NO: 206, or SEQ ID NO: 206; or sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 169, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 205, SEQ ID NO: 206, or SEQ ID NO: 206.

[0249] In some embodiments, the tumour-inducible promoter is a hypoxia-, N-acetylglucosamine, and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 204.

[0250] The hypoxia-, N-acetylglucosamine, and lactate-inducible promoter may be activated and drive expression of the extracellular matrix degrading (ECM) degrading enzyme in the presence of hypoxic conditions as described herein, N-acetylglucosamine, and lactate.

[0251] In some embodiments, the tumour-inducible promoter is a hypoxia-, cAMP, and lactate-inducible promoter. In some embodiments, the hypoxia-, cAMP, and lactate-inducible promoter comprises an FNR binding motif and / or an fnrS promoter or subsequence thereof and / or a grcA promoter or subsequence thereof, CRP binding motif, and an IldR binding motif. In some embodiments:i) a) the FNR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244; and / or

[0252] b) the fnrS promoter or subsequence thereof comprises or consists of a nucleotide sequence of SEQ ID NO: 250 or SEQ ID NO: 257, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 250 or SEQ ID NO: 257; and / or

[0253] c) the grcA promoter or subsequence thereof comprises or consists of a nucleotide sequence of SEQ ID NO: 253 or SEQ ID NO: 254, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249;

[0254] and / or

[0255] ii) the CRP binding motif comprises or consists of a nucleotide nucleotide sequence of SEQ ID NO: 245 or SEQ ID NO: 246; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 245 or SEQ ID NO: 246; and / or

[0256] iii) the lldR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 248, SEQ ID NO: 249, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249.

[0257] In some embodiments, the hypoxia-, cAMP, and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 170, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 189, SEQ ID NO: 208, SEQ ID NO: 209, or SEQ ID NO: 210; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 170, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 189, SEQ ID NO: 208, SEQ ID NO: 209, or SEQ ID NO: 210.

[0258] In some embodiments, the hypoxia-, cAMP, and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 170 or SEQ ID NO: 209; or a sequencethat is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 170 or SEQ ID NO: 209.

[0259] In some embodiments, the hypoxia-, cAMP, and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 170. In some embodiments, the hypoxia-, cAMP, and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 209.

[0260] The hypoxia-, cAMP, and lactate-inducible promoter may be activated and drive expression of the extracellular matrix degrading (ECM) degrading enzyme in the presence of hypoxic conditions as described herein, cAMP, and lactate.

[0261] In some embodiments, the tumour-inducible promoter is an ethanolamine- and lactate-inducible promoter. In some embodiments, the ethanolamine- and lactate-inducible promoter comprises a EutR binding motif and an lldR binding motif. In some embodiments: i) the EutR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 261 or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 261; and / or

[0262] ii) the lldR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 248, SEQ ID NO: 249, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249.

[0263] In some embodiments, the ethanolamine- and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, or SEQ ID NO: 196; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, or SEQ ID NO: 196.

[0264] In some embodiments, the ethanolamine- and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 193 or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 193.The ethanolamine and lactate-inducible promoter may be activated and drive expression of the extracellular matrix degrading (ECM) degrading enzyme in the presence of both ethanolamine and lactate.

[0265] In some embodiments, the tumour-inducible promoter is an ethanolamine-, lactate-, and N-acetylglucosamine-inducible promoter. In some embodiments, the ethanolamine-, lactate-, and N-acetylglucosamine-inducible promoter comprises a EutR binding motif, an IldR binding motif, and a NagC binding motif. In some embodiments:

[0266] i) the EutR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 261 or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 261;

[0267] ii) the IldR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 248, SEQ ID NO: 249, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249; and / or

[0268] iii) the NagC binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 239, or SEQ ID NO: 259, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 239, or SEQ ID NO: 259.

[0269] In some embodiments, the ethanolamine-, lactate-, and N-acetylglucosamine-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 194 or SEQ ID NO: 196; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 194 or SEQ ID NO: 196.

[0270] In some embodiments, the ethanolamine-, lactate-, and N-acetylglucosamine-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 194; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 194.

[0271] The ethanolamine-, lactate-, and N-acetylglucosamine-inducible promoter may be activated and drive expression of the extracellular matrix degrading (ECM) degradingenzyme in the presence of ethanolamine-, lactate-, and N-acetylglucosamine or N-acetylglucosamine-6-phosphate.

[0272] In some embodiments, the tumour-inducible promoter is an ethanolamine-, lactate-, and hypoxia-inducible promoter. In some embodiments, the ethanolamine-, lactate-, and hypoxia-inducible promoter comprises a EutR binding motif, an IldR binding motif, and an FNR binding motif and / or an fnrS promoter or subsequence thereof and / or a grcA promoter or subsequence thereof. In some embodiments:

[0273] i) the EutR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 261 or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 261;

[0274] ii) the IldR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 248, SEQ ID NO: 249, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249; and / or

[0275] iii) a) the FNR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244; and / or

[0276] b) the fnrS promoter or subsequence thereof comprises or consists of a nucleotide sequence of SEQ ID NO: 250 or SEQ ID NO: 257, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 250 or SEQ ID NO: 257; and / or

[0277] c) the grcA promoter or subsequence thereof comprises or consists of a nucleotide sequence of SEQ ID NO: 253 or SEQ ID NO: 254, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249.

[0278] In some embodiments, the ethanolamine-, lactate-, and hypoxia-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 195 or SEQ ID NO: 196; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%,at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 195 or SEQ ID NO: 196.

[0279] In some embodiments, the ethanolamine-, lactate-, and hypoxia-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 195; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 195.

[0280] The ethanolamine-, lactate-, and hypoxia-inducible promoter may be activated and drive expression of the extracellular matrix degrading (ECM) degrading enzyme in the presence of ethanolamine-, lactate-, and hypoxic conditions as described herein.

[0281] In some embodiments, the tumour-inducible promoter is an ethanolamine-, lactate-, hypoxia-, and N-acetylglucosamine-inducible promoter. In some embodiments, the ethanolamine-, lactate-, hypoxia- and N-acetylglucosamine-inducible promoter comprises a EutR binding motif, an lldR binding motif, an FNR binding motif and / or an fnrS promoter or subsequence thereof and / or a grcA promoter or subsequence thereof, and a NagC binding motif. In some embodiments:

[0282] i) the EutR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 261 or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 261;

[0283] ii) the lldR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 248, SEQ ID NO: 249, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249;

[0284] iii) a) the FNR binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244; and / or

[0285] b) the fnrS promoter or subsequence thereof comprises or consists of a nucleotide sequence of SEQ ID NO: 250 or SEQ ID NO: 257, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%,at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 250 or SEQ ID NO: 257;

[0286] c) the grcA promoter or subsequence thereof comprises or consists of a nucleotide sequence of SEQ ID NO: 253 or SEQ ID NO: 254, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249;

[0287] and / or

[0288] iv) the NagC binding motif comprises or consists of a nucleotide sequence of SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 239, or SEQ ID NO: 259, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 239, or SEQ ID NO: 259.

[0289] In some embodiments, the ethanolamine-, lactate-, hypoxia-, and N-acetylglucosamine-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 196; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 196.

[0290] In some embodiments, the tumour-inducible promoter provided herein comprises: a discriminator sequence (also known as a discriminator); a -10 / -35 spacer sequence (also known as a spacer or -10 / -35 spacer); a TUSP binding site; a TGIF sequence; a terminator sequence; a ribosome binding site (RBS); and / or any combination thereof.

[0291] In some embodiments, the tumour-inducible promoter provided herein comprises a discriminator sequence (also known as a discriminator). In some embodiments, the discriminator comprises or consists of a nucleotide sequence of SEQ ID NO: 229, SEQ ID NO: 230, SEQ ID NO: 231, SEQ ID NO: 232, SEQ ID NO: 233, SEQ ID NO: 234, SEQ ID NO: 235, SEQ ID NO: 236, SEQ ID NO: 237, SEQ ID NO: 238, SEQ ID NO: 240, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 229, SEQ ID NO: 230, SEQ ID NO: 231, SEQ ID NO: 232, SEQ ID NO: 233, SEQ ID NO: 234, SEQ ID NO: 235, SEQ ID NO: 236, SEQ ID NO: 237, SEQ ID NO: 238, or SEQ ID NO: 240. In some embodiments, the discriminator is a "high GC" discriminator. In some embodiments, the "high GC" discriminator has a sequence of SEQ ID NO: 232 or SEQ ID NO: 233.In some embodiments, the tumour-inducible promoter provided herein comprises a -10 / -35 spacer. In some embodiments, the -10 / -35 spacer is selected from the group comprising or consisting of: SEQ ID NO: 217, SEQ ID NO: 218, SEQ ID NO: 219, SEQ ID NO: 220, SEQ ID NO 221, SEQ ID NO: 222, SEQ ID NO: 223, SEQ ID NO: 224, SEQ ID NO: 225, or SEQ ID NO: 226, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 217, SEQ ID NO: 218, SEQ ID NO: 219, SEQ ID NO: 220, SEQ ID NO 221, SEQ ID NO: 222, SEQ ID NO: 223, SEQ ID NO: 224, SEQ ID NO: 225, or SEQ ID NO: 226.

[0292] In some embodiments, the tumour-inducible promoter provided herein comprises a TGIF sequence. TGIF motifs have been identified in hypoxia-inducible promoters. Accordingly, in some embodiments, the tumour-inducible promoter is a hypoxia inducible promoter comprising a TGIF sequence. In some embodiments, the TGIF sequence comprises or consists of a nucleotide sequence of: SEQ ID NO: 230, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 230.

[0293] In some embodiments, the tumour-inducible promoter provided herein comprises a tumour-specific motif (TUSP). Tumour-specific motifs have been identified in tumourinducible promoters. In some embodiments, the tumour-specific motif (TUSP) comprises or consists of a nucleotide sequence of: SEQ ID NO: 227, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 227

[0294] In some embodiments, the tumour-inducible promoter comprises a terminator sequence. In some embodiments, the terminatory sequence comprises or consists of a nucleotide sequence of: SEQ ID NO: 247, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 247.

[0295] In some embodiments, the tumour inducible promoter comprises a ribosome binding site (RBS). In some embodiments, the ribosome binding site (RBS) is encoded by a nucleotide sequence of SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO:88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, or SEQ ID NO: 105; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, or SEQ ID NO: 105.

[0296] In some embodiments, the tumour-inducible promoter does not comprise or consist of a sequence of SEQ ID NO: 271, SEQ ID NO: 272, SEQ ID NO: 273, SEQ ID NO: 274.

[0297] As disclosed herein, the promoter elements may be arranged in a variety of different ways, and the sequences of each promoter element varied.

[0298] The tumour-inducible promoter provided herein comprises a modified regulatory element selected from the group comprising or consisting of: a modified RNA polymerase binding site, a modified ribosome binding site (RES), a modified -35 sequence, a modified -10 sequence, a modified Shine-Dalgarno sequence, a modified Pribnow Box, modified operator sequence, a modified TATA box, a modified transcription factor binding site, a modified discriminator sequence, a modified -10 / -35 spacer sequence; or any combination thereof.

[0299] A promoter may be identified as a pH-inducible promoter, a hypoxia-inducible promoter, an N-acetylglucosamine-inducible promoter, an ethanolamine-inducible promoter, and / or a lactate-inducible promoter using methods known in the art. Exemplary methods of identifying a promoter as pH-inducible are disclosed in Chen et al., 2018, " Tuning the dynamic range of bacterial promoters regulated by ligand-inducible transcription factors", Nat. Commun., 9:64 (herein incorporated by reference in its entirety). For example, a cell may be provided comprising a nucleic acid comprising a nucleotide sequence encoding a promoter sequence operably linked to a nucleotide sequence encoding a detectable marker, for example a detectable marker that is capable of producing light. Suitabledetectable markers may include, for example, fluorescent proteins (including but not limited to: GFP, YFP, RFP, mCherry, BFP, iLOV and any variant or derivative thereof), luciferase, horseradish peroxidase, alkaline phosphatase, and the like. The cell may be cultured in suitable test conditions, for example in culture media that has a test pH, that is hypoxic and / or has a hypoxic O2concentration as discussed herein, that comprises N-acetylglucosamine, that comprises N-acetylglucosamine-6-phosphate, that comprises ethanolamine, that comprises adenosylcobalamin, that comprises cAMP, and / or that comprises lactate. Suitable controls may be employed, for example culturing the cell in suitable control conditions. In the case of identifying pH-inducible, hypoxia-inducible, N-acetylglucosamine-inducible, ethanolamine-inducible, cAMP-inducible, and / or lactate-inducible promoters, the control condition may culturing the cell in culture media that has a pH that is different from the test pH, in media that is normoxic or not hypoxic and / or has a normoxic O2concentration as discussed herein, in media that does not comprise or comprises a lower concentration of N-acetylglucosamine, ethanolamine, adenosylcobalamin, cAMP, or lactate than the concentration of N-acetylglucosamine, ethanolamine, adenosylcobalamin, cAMP, or lactate in the test condition. The presence or absence of a signal (for example, light of a given wavelength) by the cell may then be detected in the test conditions and / or the control conditions. Where the signal is detected in the test conditions, the promoter is determined to be induced in said test conditions -for example, the promoter may be determined to be a pH-inducible promoter, a hypoxiainducible promoter, an N-acetylglucosamine-inducible promoter, an ethanolamine-inducible promoter, a cAMP-inducible promoter, and / or a lactate-inducible promoter. Similarly, where suitable controls are employed, where the signal is detected in the test conditions but is not detected in the control conditions or is detected at a statistically significantly lower level in the control conditions than in the test conditions, the promoter is determined to be induced in said test conditions - for example, the promoter may be determined to be a pH-inducible promoter, a hypoxia-inducible promoter, an N-acetylglucosamine-inducible promoter, an ethanolamine-inducible promoter, a cAMP-inducible promoter, and / or a lactate-inducible promoter. The signal may be present at a level that is 2x, 3x, 4x, 5x, 6x, 7x, 8x, 9x, 10x, 20x, 30x, 40x, 50x, 60x, 70x, 80x, 90x, 100x, 200x, 300x, 400x, 500x, 600x, 700x, 800x, 900x, 1000x, 2000x, 3000x, 4000x, 5000x, 6000x, 7000x, 8000x, 9000x, 10000x, or more times higher in the test conditions than in the control conditions. Suitable methods of detecting a detectable marker are known in the art, and include, for example, visual inspection, light microscopy, fluorescence microscopy, flow cytometry such as fluorescence activated cell sorting (FACS), and microplate readers.In some embodiments, the tumour-inducible promoter is dual-inducible promoter that is hypoxia- and pH-inducible as disclosed in International Patent Application No. PCT / GB2024 / 051897. For example, a dual inducible promoter may be induced in hypoxic conditions and may have increased activity at a given pH, for example at a pH that is an acidic pH as provided herein.

[0300] Accordingly, in some embodiments, the dual inducible promoter is a hypoxia-inducible promoter the activity of which is increased in acidic conditions. In some embodiments, the dual inducible promoter is activated and drives expression of the heterologous ECM degrading polypeptide at an oxygen concentration of:

[0301] a) between about 0% and about 2%, about 0.2% and about 1.8%, about 0.4% and about 1.6%, about 0.6% and about 1.4%, about 0.8% and about 1.2%, about 0% and about 1%, about 0.2% and about 0.8%, about 0.4 and about 0.6%, 1% and 2%, about 1.2% and about 1.8%, about 1.4% and about 1.6%;

[0302] b) between 0% and 2%, 0.2% and 1.8%, 0.4% and 1.6%, 0.6% and 1.4%, 0.8% and 1.2%, 0% and 1%, 0.2% and 0.8%, 0.4 and 0.6%, 1% and 2%, 1.2% and 1.8%, 1.4% and 1.6%;

[0303] c) below about 2%, below about 1.5%, below about 1.6%, below about 1.4%, below about 1.2%, below about 1%, below about 0.8%, below about 0.6%, below about 0.4%, below about 0.2% or lower;

[0304] d) below 2%, below 1.5%, below 1.6%, below 1.4%, below 1.2%, below 1%, below 0.8%, below 0.6%, below 0.4%, below 0.2% or lower;

[0305] e) about 2%, about 1.8%, about 1.6%, about 1.4%, about 1.2%, about 1%, about 0.8%, about 0.6%, about 0.4%, about 0.2%; and / or

[0306] f) 2%, 1.8%, 1.6%, 1.4%, 1.2%, 1%, 0.8%, 0.6%, 0.4%, 0.2%.

[0307] In some embodiments, the dual-inducible promoter is not activated and does not drive expression of the heterologous ECM degrading polypeptide in normoxia; optionally wherein the dual-inducible promoter is not activated and does not drive expression of the heterologous ECM degrading polypeptide at an oxygen concentration of:

[0308] a) above about 6%, above about 5%, above about 4%, or above about 3%;

[0309] b) above 6%, above 5%, above 4%, or above 3%;

[0310] c) about 6%, about 5%, about 4%, about 3%; and / or

[0311] d) 6%, 5%, 4%, 3%.

[0312] In some embodiments, the dual-inducible promoter is activated and drives expression and / or increased expression of the heterologous ECM degrading polypeptide at an acidic pH; optionally wherein the acidic pH is:a) between about pH 5.5 and about pH 7, about pH 5.6 and about pH 6.9, about pH 5.7 and about pH 6.8, about pH 5.8 and about pH 6.7, about pH 5.9 and about pH 6.6, about pH 6.0 and about pH 6.5, about pH 6.1 and about pH 6.4, about pH 6.2 and about pH 6.3;

[0313] b) between pH 5.5 and pH 7, pH 5.6 and pH 6.9, pH 5.7 and pH 6.8, pH 5.8 and pH 6.7, pH 5.9 and pH 6.6, pH 6.0 and pH 6.5, pH 6.1 and pH 6.4, pH 6.2 and pH 6.3 c) below about pH 7, below about pH 6.9, below about pH 6.8, below about pH 6.7, below about pH 6.6, below about pH 6.5, below about pH 6.4, below about pH 6.3, below about pH 6.2, below about pH 6.1, below about pH 6.0, below about pH 5.9, below about pH 5.8, below about pH 5.7, below about pH 5.6, or lower;

[0314] d) below pH 7, below pH 6.9, below pH 6.8, below pH 6.7, below pH 6.6, below pH 6.5, below pH 6.4, below pH 6.3, below pH 6.2, below pH 6.1, below pH 6.0, below pH 5.9, below pH 5.8, below pH 5.7, below pH 5.6;

[0315] e) about pH 5.5, about pH 5.6, about pH 5.7, about pH 5.8, about pH 5.9, about pH 6.0, about pH 6.1, about pH 6.2, about pH 6.3, about pH6.4, about pH 6.5, about pH 6.6, about pH 6.7, about pH 6.8, about pH 6.9, or about pH 7; and / or

[0316] f) pH 5.5, pH 5.6, pH 5.7, pH 5.8, pH 5.9, pH 6.0, pH 6.1, pH 6.2, pH 6.3, pH6.4, pH 6.5, pH 6.6, pH 6.7, pH 6.8, pH 6.9, or pH 7.

[0317] In some embodiments, the dual-inducible promoter is activated and drives expression and / or increased expression of the heterologous ECM degrading polypeptide at an acidic pH; optionally wherein the acidic pH is:

[0318] a) between about pH 5.9 and about pH 6.5, about pH 5.95 and about pH 6.45, about pH 6.0 and about pH 6.4, about pH 6.05 and about pH 6.35, about pH 6.1 and about 6.3, or about pH 6.15 and about pH 6.25;

[0319] b) between pH 5.9 and pH 6.5, pH 5.95 and pH 6.45, pH 6.0 and pH 6.4, pH 6.05 and pH 6.35, pH 6.1 and 6.3, or pH 6.15 and pH 6.25;

[0320] c) about pH 5.9, about pH 5.95, about pH 6.0, about pH 6.05, about pH 6.1, about pH 6.15, about pH 6.2, about pH 6.25, about pH 6.3, about pH 6.35, about pH 6.4, about pH 6.45, or about pH 6.5; and / or

[0321] d) pH 5.9, pH 5.95, pH 6.0, pH 6.05, pH 6.1, pH 6.15, pH 6.2, pH 6.25, pH 6.3, pH 6.35, pH 6.4, pH 6.45, or pH 6.5.

[0322] In some embodiments, the dual-inducible promoter is LOR9 or LOR1. In some embodiments, the dual-inducible promoter is encoded by a nucleotide sequence of SEQ ID SEQ ID NO: 55, or SEQ ID NO: 56; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%,at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 55, or SEQ ID NO: 56.

[0323] Dual-inducible promoters may be identified using the same methods as set out for identifying pH-inducible and hypoxia-inducible promoters. If a promoter is identified as a pH-inducible promoter and a hypoxia-inducible promoter, it may be identified as a dualinducible promoter. If a promoter is identified as a hypoxia-inducible promoter, and the activity of the hypoxia-inducible promoter increases in conditions that are the same as the conditions that identify a promoter as a pH-inducible promoter, the promoter may be determined to be a dual-inducible promoter.

[0324] In some embodiment, the promoter may be a hybrid promoter. Transcription from a hybrid promoter may be activated by two or more transcription factors, either separately or concurrently. Hybrid promoters may be constructed, for example, by encoding operator sites for multiple transcription factors in a single promoter; operably linking a gene to two consecutive promoters, each comprising specific regulatory features and transcription start sites; or by engineering different transcription factors that bind to the same operator.

[0325] Optimal activity of the heterologous extracellular matrix (ECM) degrading polypeptide may depend on the localisation of the heterologous extracellular matrix (ECM) degrading polypeptide with respect to the recombinant cell. For example, expression of or localisation of the extracellular matrix (ECM) degrading polypeptide in the cell or display of the extracellular matrix (ECM) degrading polypeptide on the surface of the cell may restrict degradation of extracellular matrix (ECM) to the vicinity of the cell, providing controlled degradation of extracellular matrix (ECM) by the cell. Secretion of the extracellular matrix (ECM) degrading polypeptide from the cell may allow the extracellular matrix (ECM) degrading polypeptide to degrade extracellular matrix (ECM) that is not proximal to or contacted by the cell. Without wishing to be bound by theory, it is thought that extracellular matrix (ECM) degrading polypeptide secreted from the cell may diffuse through extracellular matrix (ECM) or a tumour, allowing it to degrade extracellular matrix (ECM) in portions of the tumour that are not immediately contacted by the cell. It is expected that the polypeptide may not diffuse outside of the tumour (for example, due to the size and / or short half-life of the polypeptide), may not retain extracellular matrix (ECM) degrading activity outside of the tumour, and / or may not diffuse outside of the tumour in an amount, quantity, or concentration to effectively degrade the extracellular matrix (ECM) present in tissues outside of the tumour.

[0326] In some embodiments, the heterologous extracellular matrix (ECM) degrading polypeptide:a) is secreted from the cell;

[0327] b) is displayed on the surface of the cell; or

[0328] c) is expressed in the cell.

[0329] In a preferred embodiment, the heterologous extracellular matrix (ECM) degrading polypeptide is secreted from the cell.

[0330] By "secreted by the cell" it is meant actively exported by the cell across the plasma membrane or plasma membranes of the cell, from the cytosol to the extracellular milieu, via a polypeptide secretion system that is expressed by the cell. The polypeptide secretion system may be endogenous to the cell or may be introduced to the cell. Suitable polypeptide secretion systems may be selected from the group comprising or consisting of: a type 1 secretion system (T1SS), a type 2 secretion system (T2SS), a type 3 secretion system (T3SS), a flagellar type 3 secretion system (F-T3SS), a non-flagellar type 3 secretion system (NF-T3SS), a type 4 secretion system (T4SS), a type 5 secretion system, a type 5 secretion system, a type 6 secretion system (T6SS), a type 7 secretion system (T7SS), a type 8 secretion system (T8SS), a type 9 secretion system (T9SS), a type 10 secretion system (T10SS), a type 11 secretion system (T11SS), an autotransporter, a Usp45-mediated secretion mechanism, a Sec dependent secretion system, an ompin-dependent secretion system, a Tat dependent secretion system, a Lactococcus secretion system, or any combination thereof. Suitable secretion systems and cognate signal sequences capable of directing a polypeptide for secretion by these secretion systems are disclosed in WO 2021 / 255480 Al, Burdette et al., 2018, " Developing Gram-negative bacteria for the secretion of heterologous proteins" Microb. Cell Factories, 17:196, Costa et al., 2015, " Secretion systems in Gram-negative bacteria: structural and mechanistic insights", Nat. Rev. Microbiol., 13:343-359, Zhang etal., 2006, " Extracellular accumulation of recombinant proteins fused to the carrier protein YebF in Escherichia coli", Nat. Biotechnol., 24:100-104, Borrero et al., 2011, " Use of the usp45 lactococcal secretion signal sequence to drive the secretion and functional expression of enterococcal bacteriocins in Lactococcus lactis", Appl. Microbiol. Biotechnol., 89(1): 131-143, and Kim et al., 2014, " Simple amino acid tags improve both expression and secretion of Candida antarctica lipase B in recombinant Escherichia coli", Biotechnol. Bioeng., 112(2):346-355, each of which is herein incorporated by reference in its entirety.

[0331] As is known in the art, secretion of a polypeptide from a bacterial cell via a particular secretion system requires the polypeptide to comprise a secretion signal or secretion domain that targets the polypeptide to said particular secretion system. However, such secretion signals or secretion domains have degenerate sequences; and accordingly, asecretion signal or secretion domain that is not 100% identical to the consensus sequence or a known sequence of said secretion signal or secretion domain may still be secreted by the target secretion system. For example, a secretion signal or secretion domain may be at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the consensus sequence or known sequence of a secretion signal or domain and be secreted by the target secretion system.

[0332] Suitable secretion signals or secretion domains may be identified by methods known in the art. Such methods may include, for example, culturing in test conditions a bacterial cell expressing a fusion polypeptide comprising the secretion signal or secretion domain and a second polypeptide of interest in liquid culture media; separating the bacterial cell from the liquid culture media; and identifying the presence or absence of the fusion polypeptide or second polypeptide of interest in the separated culture media. Suitable controls may be employed, for example separately culturing a bacterial cell expressing a control polypeptide comprising the second polypeptide but that does not comprise the secretion signal or secretion domain; separating the bacterial cell from the liquid culture media; and identifying the presence or absence of the control polypeptide in the separated culture media. A further suitable control may be used where the fusion polypeptide is placed under the control of an inducible promoter. Such a control may include separately culturing a bacterial cell expressing a control polypeptide comprising the secretion signal or secretion domain and second polypeptide; separating the bacterial cell from the liquid culture media; and identifying the presence or absence of the control polypeptide in the separated culture media. Where the fusion polypeptide or second polypeptide of interest is present in the separated culture media of the test conditions, the fusion polypeptide or second polypeptide of interest is determined to be secreted by the bacterial cell. Similarly, where suitable controls are employed, where the fusion polypeptide or second polypeptide of interest is present in the separated culture media of the test conditions, but the control polypeptide is absent from the control conditions or is present at a statistically significantly lower level in the control conditions than in the test conditions, the fusion polypeptide or second polypeptide of interest is determined to be secreted by the bacterial cell. The fusion polypeptide or second polypeptide of interest may be present in the test condition at a level that is 2x, 3x, 4x, 5x, 6x, 7x, 8x, 9x, 10x, 20x, 30x, 40x, 50x, 60x, 70x, 80x, 90x, 100x, 200x, 300x, 400x, 500x, 600x, 700x, 800x, 900x, 1000x, 2000x, 3000x, 4000x, 5000x, 6000x, 7000x, 8000x, 9000x, 10000x, or more times higher than the control polypeptide is present in the control conditions. Methods of detecting protein in separated liquid culture media are known to the person skilled in the art, and include, for example, western blot analysis, HPLC analysis, ELISA analysis, and the like.Methods of separating bacterial cells from liquid culture media or liquid media are known to the person skilled in the art, and include - for example - centrifugation and filtration. For example, in some embodiments, bacterial cells may be separated from liquid culture media or liquid media by centrifugation at 4000 rpm, 5000 rpm, 6000 rpm, 7000 rpm, 8000 rpm, 9000 rpm, 10,000 rpm, 11,000 rpm, 12,000 rpm, 13,000 rpm or more for a specified period. In some embodiments, bacterial cells may be separated from liquid culture media or liquid media by centrifugation for 1 minute, 2 minutes, 3 minutes, 4 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, or more. In some embodiments, bacterial cells may be separated from liquid culture media or liquid media by filtration. As the skilled person will be aware, bacterial cells from different bacterial species have different sizes. For example, an Escherichia coli cell has a diameter of approximately 0.25 to 1 pm and a length of 1-2 pm; whereas a Lactococcus lactis cell has a length of approximately 0.5-1.5 pm and a diameter of approximately 0.7-1 pm. Appropriate filter sizes for separating bacterial cells from liquid culture media or liquid media may be determined by the person skilled in the art.

[0333] Exemplary secretion domains and the exemplary secretion system via which a polypeptide comprising said secretion domain may be exported from the cell are set out in Table 1.

[0334] Table 1

[0335] Secretion system Secretion domain

[0336] Sec-dependent secretion system N-terminal PelB sequence

[0337] Sec-dependent secretion system N-terminal YebF carrier protein Autotransporter N-terminal CtxB signal sequence

[0338] C-terminal AIDA-I autotransporter Flagellar T3SS N-terminal FliC signal sequence Lactococcus secretion system N-terminal Usp45 signal sequence

[0339]

[0340] Such secretion domains may be used to target a fusion polypeptide for secretion by the cell. Accordingly, in some embodiments, the heterologous extracellular matrix (ECM) degrading polypeptide is a fusion polypeptide comprising:

[0341] a) an ECM degrading domain; and

[0342] b) a secretion domain.

[0343] In some embodiments, the heterologous extracellular matrix (ECM) degrading polypeptide comprises two secretion domains.In some embodiments, the secretion domain is selected from the group comprising or consisting of: a PelB secretion signal sequence, a YebF carrier protein, a CtxB signal sequence, an AIDA-I autotransporter, a FliC signal sequence, and a Usp45 signal sequence, or any combination thereof. As will be appreciated, where secretion of the polypeptide requires the presence of a particular cognate secretion system (for example, the FliC signal sequence and the T3SS), a cell should be selected comprising said secretion system. As described herein, however, the polypeptide secretion system may be endogenous to the cell or may be introduced to the cell, using methods known in the art.

[0344] In some embodiments, the secretion domain:

[0345] a) comprises or consists of an amino acid sequence of SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35; or of an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35; and / or

[0346] b) is encoded by a nucleotide sequence of SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36.

[0347] In some embodiments, the fusion polypeptide:

[0348] a) comprises an ECM degrading domain that comprises a hyaluronidase, and:

[0349] i) comprises or consists of an amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45; or of an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 1, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45; and / or

[0350] ii) is encoded by a nucleotide sequence of SEQ ID NO: 2, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ IDNO: 2, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46; or

[0351] b) comprises an ECM degrading domain that comprises a neuraminidase, and:

[0352] i) comprises or consists of an amino acid sequence of SEQ ID NO: 5, SEQ ID NO: 106, SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 114, or an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 5, SEQ ID NO: 106, SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 112, or SEQ ID NO: 114; and / or

[0353] ii) is encoded by a nucleotide sequence of SEQ ID NO: 6, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 6, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, or SEQ ID NO: 115.

[0354] In some embodiments, the secretion domain is not PhoA. In some embodiments, the secretion domain is not OmpF. In some embodiments, the secretion domain is not CvaC. In some embodiments, the secretion domain is not TorA. In some embodiments, the secretion domain is not FdnG. In some embodiments, the secretion domain is not DmsA. In some embodiments, the secretion domain is not HlyA.

[0355] In some embodiments, the heterologous extracellular matrix (ECM) degrading polypeptide comprises a cleavage domain. By the term "cleavage domain" it is meant a portion of a polypeptide that is capable of being cleaved or split by a polypeptide-degrading polypeptide, such as a protease or a peptidase (for example, an endopeptidase or exopeptidase). Cleavage of the cleavage domain may be site- or sequence-specific, for example the cleavage domain may comprise an amino acid sequence that is or is capable of being recognised and cleaved or split by a specific protease or peptidase that is capable of cleaving or splitting said amino acid sequence. Such an amino acid sequence may be referred to as a "protease site" or a "peptidase site".

[0356] In some embodiments, the cleavage domain is positioned between the extracellular matrix (ECM) degrading domain and the secretion domain, thereby allowing cleavage of the extracellular matrix (ECM) degrading domain from the secretion domain. It is thought that cleavage of the ECM degrading domain from the secretion domain at the cleavage domain may improve secretion of the extracellular matrix (ECM) degrading polypeptide from the cell. Without wishing to be bound by theory, some secretion domains as provided hereinare known to associate with the outer membrane or outer membrane proteins of a cell during the secretion of cargo proteins fused to said secretion domains; and it is thought that the cleavage of a cargo protein (for example, an extracellular matrix (ECM) degrading domain as provided herein) from the secretion domain may increase release of the extracellular matrix (ECM) degrading polypeptide from the surface of the cell into the extracellular milieu.

[0357] In some embodiments, the cleavage domain comprises an amino acid sequence that is or is capable of being recognised and cleaved or split by a protease or peptidase that is present in a target tissue or tumour as described herein, for example a protease or peptidase that is produced naturally by a human subject. Human proteases and peptidases are known in the art, and may be selected from the groups comprising: a meta I loprotease (including a matrix metalloprotease (MMP)), a serine protease, a cysteine protease, a threonine protease, an aspartic protease, a glutamic protease, an asparagine protease, and a peptide lyase. In some embodiments, the cleavage domain comprises an amino acid sequence that is or is capable of being recognised and cleaved or split by a matrix metalloprotease (MMP), i.e., the cleavage domain comprises a matrix meta I loprotease (MMP) site.

[0358] It may be advantageous to provide a nucleotide sequence encoding a transcriptional regulatory polypeptide, which is capable of modulating transcription from the promoter in response to the presence of a given molecule or certain conditions. Accordingly, in one embodiment, the nucleic acid further comprises nucleotide sequence encoding a transcriptional regulatory polypeptide.

[0359] The transcriptional regulatory polypeptide may suitably be a transcriptional repressor, a transcriptional activator, an operator protein, or any combination thereof. The transcriptional regulatory polypeptide may be functionally related to the secretion signal or secretion domain. Accordingly, in some embodiments, the secretion domain is a FliC signal sequence, the transcriptional regulatory peptide is selected from the group comprising or consisting of: a FliC repressor; optionally wherein the FliC repressor is a GadE repressor. It is thought that GadE represses fliC transcription by binding directly to the fliC promoter to repress transcription. GadE expression should interrupt the flagellin protein to be secreted by the T3SS and to compete with the heterologous extracellular matrix (ECM) degrading polypeptide secretion.

[0360] In some embodiments, the transcriptional regulatory peptide:a) comprises or consists of an amino acid sequence of SEQ ID NO: 49; or of an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 49; and / or

[0361] b) is encoded by a nucleotide sequence of SEQ ID NO: 50, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 50.

[0362] In some embodiments, the transcriptional regulatory peptide is selected from the group comprising or consisting of FNR; NagC; CRP; EutR; and / or LldR.

[0363] The recombinant bacterial cell may be any suitable bacterial cell. However, the cell is advantageously a bacterial cell that is genetically tractable; actively migrates to tumours; and has an intrinsic ability to trigger an immune response within a tumour. Because of the infection risks and side-effects (for example, systemic inflammation, toxic shock syndrome, and the like) associated with administering a pathogenic cell to a subject, it is considered advantageous if the cell is not a pathogenic cell or is a non-pathogenic cell. By "pathogenic cell" it is meant a cell that is capable of causing a disease in a subject, for example by presenting virulence factors such as persistence in immune cells. In some embodiments, therefore, the cell is a non-pathogenic bacterial cell, a commensal cell, or a probiotic cell. As is known in the art, a commensal cell is a cell that is a constituent of the healthy microbiome of a subject, for example on the surface of a subject's body or mucous membranes, and which does not usually cause disease or infection in a healthy subject. Accordingly, a commensal cell may be a constituent of the gut microbiome, the skin microbiome, the conjunctival microbiome, the urethral and bladder microbiome, the vaginal microbiome, the uterine microbiome, the oral microbiome, the nasal microbiome, the lung microbiome, and / or the biliary tract microbiome. A probiotic cell is a cell that promotes the formation or maintenance of a healthy microbiome in a subject, and does not usually cause disease or infection in a healthy subject. A probiotic cell may be a cell that is a constituent of the healthy microbiome of a subject.

[0364] As disclosed herein, the tumour environment or microenvironment may be hypoxic, anaerobic, anoxic, or microaerobic. The recombinant cell according to the present invention is therefore advantageously capable of surviving and / or replicating in a hypoxic, anaerobic, anoxic, or microaerobic environment. Thus, in some embodiments, the cell is a facultatively anaerobic cell.The recombinant bacterial cell may be any suitable bacterial cell. In some embodiments, the cell is:

[0365] a) a gram-negative bacterial cell; or

[0366] b) a gram-positive bacterial cell.

[0367] In some embodiments, the gram-negative bacterial cell is selected from the group comprising or consisting of: an Escherichia coli cell;

[0368] Optionally

[0369] wherein the gram-negative bacterial cell is an Escherichia coli K12 cell or an Escherichia coli Nissile cell;

[0370] optionally

[0371] wherein the gram-negative bacterial cell is an Escherichia coli K12 cell; optionally wherein the Escherichia coli K12 cell is selected from the group comprising or consisting of: an Escherichia coli K12 MG1655 cell, an Escherichia coli K12 58 cell, an Escherichia coli K12 679 cell, an Escherichia coli K12 WG1 cell, an Escherichia coli K12 5K cell, an Escherichia coli K12 58-161 cell, an Escherichia coli K12 AB284 cell, an Escherichia coli K12 AB311 cell, an Escherichia coli K12 AG1 cell, an Escherichia coli K12 0600 cell, an Escherichia coli K12 Cavalli Hfr cell, an Escherichia coli K12 DH1 cell, an Escherichia coli K12 DH5-0 cell, an Escherichia coli K12 DP50 cell, an Escherichia coli K12 EMG2 cell, an Escherichia coli K12 EPI100- T1R cell, an Escherichia coli K12 H1443 cell, an Escherichia coli K12 HB101 cell, an Escherichia coli K12 Hfr3000 cell, an Escherichia coli K12 Hfr 3000 X74 cell, an Escherichia coli K12 HMS 174 cell, an Escherichia coli K12 JM109 cell, an Escherichia coli K12 TGI cell, an Escherichia coli K12 TOPIO cell, an Escherichia coli K12 W1485 cell, an Escherichia coli K12 W208 cell, an Escherichia coli K12 W3110 cell, an Escherichia coli K12 W945 cell, an Escherichia coli K12 WA704 cell, and an Escherichia coli K12 WG1 cell. In some embodiments, the Escherichia coli K12 cell is an Escherichia coli K12 MG1655 cell.

[0372] E. coli K12 (such as E. coli K12 MG1655) is a non-pathogenic facultatively anaerobic rodshaped gram negative bacterium that has a good safety profile, is capable of localising to a tumour, and is genetically tractable. Considered as safe by regulators, E. coli K12 (such as E. coli K12 MG1655) has the ability to home to the tumour after intravenous administration with a high maximum tolerated dose of 5xl07CFU (Kang et al., 2020, " Imaging of tumor colonization by Escherichia coli using 18F-FDS PET", Theranostics, 2020; 10(11):4958-4966; herein incorporated by reference in its entirety).In some embodiments, the gram-positive bacterial cell is a lactic acid bacterial cell; optionally wherein the lactic acid bacterial cell is selected from the group comprising or consisting of: a Lactococcus lactis cell, a Lactobacillus cell, and a Bifidobacterium cell; optionally

[0373] wherein the gram-positive bacterial cell is a Lactococcus lactis MG1363 cell.

[0374] Lactococcus lactis MG1363, is a plasmid-free lactic acid bacteria regarded as safe by regulators. As a single-membrane gram-positive cocci, the molecular biology tools to express heterologous proteins are well defined allowing the use of the strain as a live vector to express therapeutics (Jacouton et al., 2019, " Anti-tumoral Effects of Recombinant Lactococcus lactis Strain Secreting IL-17A Cytokine", Front Microbiol., 9:3355; herein incorporated by reference in its entirety).

[0375] As discussed herein, the cell is advantageously not a pathogenic cell or is a non-pathogenic cell. Pathogenic cells may be attenuated by methods known in the art; however, the safety profile of these cells may not be comparable to that of non-pathogenic cells. Accordingly, in some embodiments, the cell is not an attenuated bacterial cell.

[0376] By "attenuated bacterial cell" it is meant a bacterial cell with reduced virulence compared to a pathogenic bacterial cell of the same strain or species that has not been "attenuated". By "attenuated" it is meant procedures that weaken an agent of disease (a pathogen). An attenuated pathogen is weakened, less vigorous compared to one that is non-attenuated. Attenuation may be due to genetic mutations. Genetic mutations may be engineered or result from passaging of the pathogen in cell culture.

[0377] Advantageously, in some embodiments, the cell is not a genotoxic cell. Advantageously, in some embodiments, the cell is a non-genotoxic cell. Advantageously, in some embodiments, the cell does not produce a genotoxin.

[0378] As used herein, the term "genotoxic" in reference to a cell indicates that the cell is capable of damaging, or that damage, a cell's genetic material (e.g., DNA), thereby causing mutations or altering its structure. As used herein, the term "genotoxin" refers to a substance or agent that is capable of damaging, or that damages, a cell's genetic material (e.g., DNA), thereby causing mutations or altering its structure. For example, a genotoxic substance or agent may be capable of damaging, or may damage, the genetic material (e.g., DNA) of a human cell comprised by the body of a subject or patient to whom the cell of the invention is administered, for example as part of a medical use or method of treatment provided herein.Genotoxins are known to the person skilled in the art, as are genotoxic cells that produce said genotoxins. Exemplary genotoxins may be selected from the group comprising or consisting of: colibactin, cytolethal distending toxin, typhoid toxin.

[0379] In some embodiments, the cell does not produce colibactin. In some embodiments, the cell is not a colibactin-producing Escherichia coli cell. Colibactin-producing cells are known to the person skilled in the art. Exemplary colibactin-producing cells may be selected from the group comprising or consisting of: Escherichia coli Nissile; Escherichia coli Nissile 1917, an Extracellular Pathogenic Escherichia coli (ExPEC). In some embodiments, the cell is not an Escherichia coli Nissile cell.

[0380] In some embodiments, the cell does not produce cytolethal distending toxin.

[0381] In some embodiments, the cell does not produce typhoid toxin.

[0382] In some embodiments, the cell does not produce a cytotoxin. In some embodiments, the cell does not produce a lytic peptide.

[0383] As used herein, the term "cytotoxin" encompasses a compound or polypeptide that capable of being toxic or causing toxicity to a cell, or is toxic to a cell, contacted with the cytotoxin; for example, a human cell comprised by the body of a subject or patient to whom the cell of the invention is administered, for example as part of a medical use or method of treatment provided herein. As used herein, the term "lytic peptide" encompasses a peptide or polypeptide that is capable of causing cell lysis, or causes cell lysis, in a cell contacted by the lytic peptide; for example, a human cell comprised by the body of a subject or patient to whom the cell of the invention is administered, for example as part of a medical use or method of treatment provided herein. Cytotoxins and lytic peptides are known to the person skilled in the art.

[0384] In some embodiments, the cell does not produce O-antigen. In some embodiments, the cell does not comprise an O-antigen. In some embodiments, the cell does not produce a bacterial capsule. In some embodiments, the cell does not comprise a bacterial capsule.

[0385] In some embodiments, the cell does not comprise an active prophage. In some embodiments, the cell comprises an inactive prophage. In some embodiments, the cell comprises a non-functional prophage.Prophages are known to the person skilled in the art. The term "prophage" includes a bacteriophage genome that is integrated into a nucleic acid (such as the bacterial genome or a plasmid) comprised by a bacterial genome. Bacteriophages can be lytic or temperate. Lytic bacteriophages infect bacterial cells and then program the synthesis of progeny bacteriophages, which are then released from the lysed cell. Temperate bacteriophages establish a stable relationship with their host bacteria in which the integrated phage DNA ( / .e., the prophage) is replicated as part of the nucleic acid comprised by the bacterial cell, and bacteriophage genes are not expressed - i.e., the prophage is not active or is inactive. However, bacteriophage particles can be released from cells containing an intact prophage by a process called induction, during which prophage genes required for lytic growth are turned on and progeny phage particles are produced and released from the cell through lysis of the cell. Not all prophages are capable of a lytic cycle. Non-functional, i.e., defective or cryptic prophages can accrue to a high level of abundancy in many bacteria as a result of mutational decay and / or the loss of one or more genes essential to the lytic cycle over thousands of bacterial replication cycles.

[0386] In some embodiments, the cell is not:

[0387] a) a Salmonella cell; optionally a Salmonella bongori cell, a Salmonella cholreaesuis cell, a Salmonella enterica cell, a Salmonella enteritidis cell, a Salmonella paratyphi cell, a Salmonella typhi cell, or a Salmonella typhimurium cell;

[0388] b) a Vibrio cell; optionally a Vibrio cholerae cell or a Vibrio fischeri cell;

[0389] c) a Shigella cell; optionally a Shigella boydii cell, a Shigella dysenteriae cell, a Shigella flexneri cell, or a Shigella sonnei cell;

[0390] d) a Lactobacillus cell; optionally a Lactobacillus bulgaricus cell or a Lactobacillus plantarum cell;

[0391] e) a Listeria cell; optionally a Listeria monocytogenes cell;

[0392] f) an Enterococcus cell; optionally an Enterococcus faecium cell;

[0393] g) a Streptococcus cell; optionally a Streptococcus pyogenes cell, a Streptococcus zooepidemic cell, or a Streptococcus pneumoniae cell;

[0394] h) a pathogenic Escherichia coli cell; optionally an Enteroinvasive Escherichia coli (EIEC) cell, an Adherent-Invasive Escherichia coli (AIEC) cell, an Enteroaggregative Escherichia coli (EAEC) cell, a Shiga Toxin-producing Escherichia coli (STEC) cell, an Enterotoxigenic Escherichia coli (ETEC) cell, an Enterohemorrhagic Escherichia coli (EHEC) cell, or an Extra intestinal Pathogenic Escherichia coli (ExPEC) cell;

[0395] i) a Bacillus cell, optionally a Bacillus CGMCC No. 5744 cell or a Bacillus subtilis; j) an Arthrobacter cell, optionally an Arthrobacter sphaeroides cell;

[0396] k) a Streptomyces cell, iptionally a Streptomyces koganeiensis cell, a Streptomyces pristinaespiralis cell, or a Streptomyces zooepidemic cell; orI) a Bacteroides cell, optionally a Bacteroides uniformis cell.

[0397] The cell provided herein is a bacterial cell. Accordingly, as will be understood, the cell is not a eukaryotic cell such as a fungal cell, a mammalian cell, or a plant cell. For example, the cell is not a Pichia cell such as a Pichia pastoris cell (also known as a Komagataella pastoris cell).

[0398] In some embodiments, the heterologous extracellular matrix (ECM) degrading polypeptide does not localise to an intracellular compartment of the cell.

[0399] In some embodiments, the heterologous extracellular matrix (ECM) degrading polypeptide does not localise to a membrane or cell wall of the cell.

[0400] It may be desirable to include transcriptional regulatory elements in the nucleic acid. Accordingly, in some embodiments, the nucleic acid comprises a nucleotide sequence encoding a transcriptional regulatory element. In some embodiments, the nucleotide sequence encoding the heterologous extracellular matrix degrading (ECM) polypeptide is operably linked to the transcriptional regulatory element. In some embodiments, the transcriptional regulatory element is selected from the group comprising or consisting of: a terminator sequence, a repressor binding sequence, an insulator, an operator sequence, a tumour-specific motif (TUSP), a -10 / -35 spacer sequence, a discriminator sequence, or any combination thereof. In some embodiments, the transcriptional regulatory element is comprised by a promoter provided herein.

[0401] In some embodiments:

[0402] a) the terminator is selected from the group comprising or consisting of: rrnB T1 / T2 and L3S2P21-F1; optionally

[0403] wherein the terminator sequence comprises or consists of a nucleotide sequence of SEQ ID NO: 51 or SEQ ID NO: 247, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 51 or SEQ ID NO: 247;

[0404] b) the operator binding sequence is selected from the group comprising or consisting of: FNR, NagC, IldR, grcA, and TGIF; optionally

[0405] wherein the operator binding sequence comprises or consists of a nucleotide sequence selected from the group comprising or consisting of SEQ ID NO: 52, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 253, SEQ ID NO: 254, SEQ ID NO: 248, SEQ ID NO: 249, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 118, SEQ I DNO: 119, SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 253, SEQ ID NO: 254, SEQ ID NO: 248, or SEQ ID NO: 249;

[0406] c) the insulator sequence is selected from the group comprising or consisting of: RiboJ; optionally

[0407] wherein the insulator sequence comprises or consists of a nucleotide sequence of SEQ ID NO: 53 or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 53;

[0408] d) the tumour-specific motif (TUSP) comprises or consists of a nucleotide sequence selected of SEQ ID NO: 227, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 227;

[0409] e) the -10 / -35 spacer sequence comprises or consists of a nucleotide sequence selected from the group comprising or consisting of: SEQ ID NO: 217, SEQ ID NO: 218, SEQ ID NO: 219, SEQ ID NO: 220, SEQ ID NO 221, SEQ ID NO: 222, SEQ ID NO: 223, SEQ ID NO: 224, SEQ ID NO: 225, or SEQ ID NO: 226, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 217, SEQ ID NO: 218, SEQ ID NO: 219, SEQ ID NO: 220, SEQ ID NO 221, SEQ ID NO: 222, SEQ ID NO: 223, SEQ ID NO: 224, SEQ ID NO: 225, or SEQ ID NO: 226;

[0410] and / or

[0411] f) the discriminator sequence comprises or consists of a nucleotide sequence selected from the group comprising or consisting of: SEQ ID NO: 229, SEQ ID NO: 230, SEQ ID NO: 231, SEQ ID NO: 232, SEQ ID NO: 233, SEQ ID NO: 234, SEQ ID NO: 235, SEQ ID NO: 236, SEQ ID NO: 237, SEQ ID NO: 238, SEQ ID NO: 240, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 229, SEQ ID NO: 230, SEQ ID NO: 231, SEQ ID NO: 232, SEQ ID NO: 233, SEQ ID NO: 234, SEQ ID NO: 235, SEQ ID NO: 236, SEQ ID NO: 237, SEQ ID NO: 238, or SEQ ID NO: 240.

[0412] Suitable nucleic acids are known in the art. In some embodiments, the nucleic acid is selected from the group comprising or consisting of: a plasmid, a phagemid, and a bacterial artificial chromosome (BAC). In some embodiments, optionally the nucleic acid is a plasmid. In some embodiments, the nucleic acid integrated into the genome of the cell. In some embodiments, the nucleic acid may comprise a detectable marker, for example an antibiotic resistance marker or a colorimetric detectable marker such as LacZ, or adetectable marker that is capable of producing light. Suitable detectable markers may include, for example, fluorescent proteins (including but not limited to: GFP, YFP, RFP, mCherry, BFP, and any variant or derivative thereof), luciferase, horseradish peroxidase, iLOV, and the like. In preferred embodiments, the nucleic acid does not comprise an antibiotic resistance marker. Accordingly, in preferred embodiments, the recombinant bacterial cell provided herein is not resistant to an antibiotic.

[0413] In some embodiments, the recombinant bacterial cell is an auxotrophic cell. In some embodiments, the cell is not capable of endogenously expressing an essential gene. As is known in the art, the term "essential gene" generally relates to a gene that is essential for the normal growth, survival, and / or metabolism of a cell. Generally, a cell that cannot express an essential gene exhibits impaired survival, growth, and metabolism. Suitable methods of preventing expression of a gene are known in the art. Exemplary method of preventing expression of a gene may be selected from the group comprising or consisting of: gene deletion, gene disruption, and gene knockdown (e.g., RIMA interference, CRISPR interference, TALEN interference, and the like). In some embodiments, the essential gene has been deleted from the genome of the cell. Suitable methods of deleting a gene from the genome of a cell are known to the person skilled in the art and include but are not limited to: homologous recombination techniques, CRISPR gene editing, TALEN gene editing, zinc finger nuclease editing, and gene knock-in techniques.

[0414] As will be understood, where an endogenous copy of an essential gene is deleted, knocked down, or disrupted, in order to survive the cell must be provided with an exogenous source of the metabolite produced by the essential gene; or a heterologous copy of the essential gene must be provided to and expressed by the cell (also known as "complementation"). Suitable methods of complementing a gene deletion are known in the art. In some embodiments, the nucleic acid further comprises a complementation cassette comprising a nucleotide sequence encoding the essential gene operably linked to a complementation promoter.

[0415] In some embodiments, the essential gene is selected from the group comprising or consisting of: cysE, glnA, ilvD, leuB, lysA, serA, metA, glyA, hisB, ilvA, pheA, proA, thrC, trpC, tyrA, thyA, uraA, dapA, dapB, dapD, dapE, dapF, flhD, metB, metC, proAB, thiL, infA, dnaN, pheS, tyrS, metG, and adk. In some embodiments, the essential gene is selected from the group comprising or consisting of: dapD; infA; or a homologue thereof. In some embodiments, the essential gene is dapD or a homologue thereof. In some embodiments, the essential gene is dapD. In some embodiments, the essential gene is infA or a homologue thereof. In some embodiments, the essential gene is infA. In someembodiments, the essential gene comprises or consists of a nucleotide sequence of: SEQ ID NO: 268 or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 268. In some embodiments, the essential gene encodes a polypeptide comprising or consisting of a sequence of: SEQ ID NO: 270 or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 270. dapD is a component of the diaminopimelic acid (DAP) pathway and is essential to maintain membrane integrity of the cell.

[0416] In some embodiments, the essential gene is not a gene selected from the group comprising or consisting of: cysE, glnA, ilvD, leuB, lysA, serA, metA, glyA, hisB, ilvA, pheA, proA, thrC, trpC, tyrA, thyA, uraA, dapA, dapB, dapE, dapF, flhD, metB, metC, proAB, thiL, dnaN, pheS, tyrS, metG, and adk. In some embodiments, the essential gene is not thyA. In some embodiments, the essential gene is not dapA.

[0417] In some embodiments, the complementation promoter is a constitutive promoter. In some embodiments, the complementation promoter is selected from the group comprising or consisting of: PdapD; lacZ, uvrA, and aroL. In some embodiments, the complementation promoter is PdapD. In some embodiments, the complementation promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 269, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 269.

[0418] Methods of introducing a nucleic acid into a cell are known in the art, and include but are not limited to: electroporation, heat-shock transformation, conjugation, or any combination thereof. Methods of integrating a nucleic acid into the genome of a cell are known in the art, and include but are not limited to: homologous recombination, non-homologous end joining (NHEJ), transposition, retrotransposition, and chromosomal editing methods such as CRISPR-Cas editing, zinc-finger nuclease (ZNF) editing, TALEN editing, and any combination thereof. Such methods are described, for example, in Sambrook, Molecular Cloning: A Laboratory Manual, 4thEd. Cold Spring Harbor, N. Y.: Cold Spring Harbor Laboratory Press, 2012.

[0419] As discussed above, bacteria - including those provided herein - are known to accumulate in a tumour. Accordingly, in some embodiments, the cell is capable of accumulating in a target tissue or tumour. By "accumulating in a target tissue or tumour" or "accumulatesin a target tissue or tumour" it is meant that the cell is present at a higher level or concentration in a target tissue or tumour compared to the area surrounding the target tissue or tumour. A cell may be determined as having "accumulated in a target tissue or tumour" if, for example, a higher number of colony forming units (CFU) of the cell are recoverable or are recovered from a given volume or mass of the target tissue or tumour than are recoverable or are recovered from an equivalent volume or mass of the area surrounding the target tissue or tumour. CFU may be determined by methods known in the art, and include plate counting methods and microscopic determination methods. Such methods may be performed in vitro or in vivo.

[0420] Where the methods are performed in vivo, the cell may be administered to a subject. The cell may be determined as having "accumulated in a target tissue or tumour" if, for example, a higher number of colony forming units (CFU) of the cell are recoverable or are recovered from a given volume or mass of the target tissue or tumour in a subject than are recoverable or are recovered from an equivalent volume or mass of a tissue or tumour that is not the target tissue or tumour in the same subject. Methods of recovering tissue or tumour samples for CFU determination are known in the art.

[0421] Similarly, a cell provided herein may comprise a detectable marker, for example a detectable marker that is capable of producing light. Suitable detectable markers may include, for example, fluorescent proteins (including but not limited to: GFP, YFP, RFP, mCherry, BFP, and any variant or derivative thereof), luciferase, horseradish peroxidase, and the like. Such a cell may be administered to a subject. The cell may be determined as having "accumulated in a target tissue or tumour" if, for example, a higher level of a signal produced by the detectable marker (for example, light of a given wavelength) is detectable or detected in the target tissue or tumour in a subject than is detectable or detected in a tissue or tumour that is not the target tissue or tumour in the same subject. Methods of detecting the accumulation of luminescent and / or fluorescent cells in a subject are known in the art, and include, for example, the IVIS Spectrum In Vivo Imaging System as described for example in Warawa and Lawrenz, 2013, " Bioluminescent Imaging of Bacteria During Mouse Infection", In: Badr, C. (eds) Bioluminescent Imaging. Methods in Molecular Biology, vol 1098. Humana Press, Totowa, NJ (herein incorporated by reference in its entirety).

[0422] As will be understood, the term "target tissue" as used herein relates to a tissue that contains a tumour or a cancer.Promoters, nucleic acids, and cells of the invention

[0423] In a second aspect, the invention provides a tumour-inducible promoter, wherein the tumour-inducible promoter is a promoter as provided herein. In some embodiments, the tumour-inducible promoter is an engineered tumour-inducible promoter.

[0424] By "engineered tumour-inducible promoter" (also referred to herein as an "engineered promoter") it is meant that the nucleotide sequence of the promoter is not a natural promoter sequence. An engineered promoter may be derived from a natural promoter, however, and may be generated by modification of any promoter element or transcriptional regulatory element described herein, for example promoter elements selected from the group comprising or consisting of: a ribosome binding site (RBS), an operator binding site, an RNA polymerase binding site, a -10 / -35 spacer sequence, and a discriminator sequence. For example, a natural promoter may be modified by replacing a promoter element or transcriptional regulatory element that is endogenous to the natural promoter with a heterologous promoter element or transcriptional regulatory element. An engineered promoter may be generated by mutagenesis of a natural promoter or another engineered promoter, for example by error-prone PCR, site-directed mutagenesis, chemical mutagenesis, or any combination thereof. An engineered promoter may be generated by altering the length and / or GC of each promoter element for example to alter translation initiation, rate, efficiency, or tropism. For example, the length and / or GC content of a -10 / -35 spacer sequence or discriminator sequence may suitably be modified to achieve a desired translation initiation, rate, efficiency, or tropism. Methods of determining whether a promoter is a tumour-inducible promoter are provided herein, and may be applied to determine whether a promoter is an engineered tumour-inducible promoter.

[0425] In a further aspect, the invention provides a nucleic acid comprising a nucleotide sequence of the tumour-inducible promoter as provided herein.

[0426] In a further aspect, the invention provides a nucleic acid comprising a nucleotide sequence encoding an extracellular matrix (ECM) degrading polypeptide. In some embodiments, the extracellular matrix (ECM) degrading polypeptide encoded by the nucleotide sequence is the heterologous extracellular matrix (ECM) degrading polypeptide provided herein. In some embodiments, the nucleic acid further comprises a promoter; optionally wherein the promoter is the promoter as provided herein. In some embodiments, the nucleotide sequence encoding the extracellular matrix (ECM) degrading polypeptide is operably linked to the promoter. In some embodiments, the nucleic acid further comprises a transcriptional regulatory element; optionally wherein the transcriptional regulatoryelement is the transcriptional regulatory element as provided herein. In some embodiments, the nucleotide sequence encoding the extracellular matrix (ECM) degrading polypeptide is operably linked to the transcriptional regulatory element.

[0427] In a further aspect, also provided herein is a nucleic acid comprising a nucleotide sequence that encodes a biosynthetic gene. In some embodiments, the biosynthetic gene is an essential gene provided herein. Accordingly, in some embodiments, the nucleic acid and biosynthetic gene encoded thereby is suitable for use in complementing an auxotrophic phenotype of a cell. In some embodiments, the essential gene is operably linked to a promoter, such as the complementation promoter provided herein.

[0428] In some embodiments, the nucleic acid of any aspect provided herein, the nucleic acid is selected from the group comprising or consisting of: a plasmid, a phagemid, and a bacterial artificial chromosome (BAC); optionally

[0429] wherein the nucleic acid is a plasmid.

[0430] In a further aspect, the invention provides a cell comprising the engineered hypoxiainducible promoter provided herein, the engineered pH-inducible promoter provided herein, and / or the nucleic acid provided herein.

[0431] In some embodiments, the nucleic acid is:

[0432] a) extrachromosomal; or

[0433] b) integrated into the genome of the cell.

[0434] In some embodiments, the cell is a recombinant bacterial cell; optionally wherein the cell is a recombinant bacterial cell provided herein. In some embodiments, the recombinant bacterial cell is an auxotrophic cell provided herein.

[0435] Medical methods and medical uses of the recombinant bacterial cell provided herein

[0436] As discussed herein, the recombinant bacterial cell of the invention may be useful in medicine, particularly in treating cancer.

[0437] Accordingly, in one aspect the invention provides the recombinant bacterial cell provided herein for use in medicine.

[0438] In a related aspect, the invention provides the recombinant bacterial cell provided herein for use as a medicament.In a further aspect, the invention provides the recombinant bacterial cell provided herein for use in treating cancer.

[0439] In a further related aspect, the invention provides use of the recombinant bacterial cell provided herein in the manufacture of a medicament for the treatment of cancer.

[0440] In one aspect, the invention provides a method of treating cancer with the recombinant bacterial cell provided herein.

[0441] In one aspect, the invention provides the cell provided herein for use in medicine.

[0442] In a related aspect, the invention provides the cell provided herein for use as a medicament.

[0443] In a further aspect, the invention provides the cell provided herein for use in treating cancer.

[0444] In a further related aspect, the invention provides use of the cell provided herein in the manufacture of a medicament for the treatment of cancer.

[0445] In one aspect, the invention provides a method of treating cancer using the cell provided herein.

[0446] In some embodiments, the medical and / or therapeutic uses and methods provided herein comprise administering an effective dose of the recombinant bacterial cell or cell to a subject. The subject may be any suitable subject, however in some preferred embodiments the subject is human. Suitably, the subject may have cancer, may have been diagnosed with cancer, may be suspected to have cancer, or may be recovering from cancer.

[0447] In some embodiments, the cancer is a tumour solid cancer. In preferred embodiments, the cancer is an immune-excluded solid tumour cancer and / or an immune-excluded tumour. As used herein, the terms "immune-excluded solid tumour cancer" and "immune-excluded tumour" are interchangeable. As discussed herein, immune-excluded tumours are characterised by a tumour microenvironment (TME) which physically and biochemically excludes immune cells from the tumour core, thereby limiting the efficacy of cancer immunotherapies. Other characteristics of immune-excluded tumours include but are not limited to: high levels of stromal deposition which prevents effective infiltration of differentsubpopulations of immune cells into the tumour microenvironment; collapsed vasculature; low levels of oxygen; an acidic pH; production of glucose; production of N-acetylglucosamine; production of ethanolamine; production of cAMP; and production of lactate.

[0448] Cancers and tumours that may suitably be treated with the recombinant bacterial cell or cell provided herein are disclosed in Wilkinson et al. (2017) Oxford Handbook of Clinical Medicine (10th Ed), Oxford University Press, and Cassidy et al. (2015) Oxford Handbook of Oncology (4thEd), Oxford University Press (each herein incorporated by reference in their entirety). In some embodiments, the cancer comprises extracellular matrix. In some embodiments, the cancer is a cancer selected from the group comprising or consisting of: triple-negative breast cancer, microsatellite stable colon cancer, and pancreatic cancer. The cancer may be a primary cancer or a secondary cancer, for example a metastatic cancer. In some embodiments, the cancer is not a cancer that does not comprise extracellular matrix. In some embodiments, the cancer is not a blood cancer. In some embodiments, the cancer is not a cancer selected from the group comprising or consisting of: leukaemia, myeloma, lymphoma.

[0449] In one aspect, the invention provides a composition comprising the recombinant bacterial cell provided herein or the cell provided herein. In some embodiments, the composition is a pharmaceutical composition. In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier, diluent, excipient, adjuvant, buffer, salt, or the like. Suitable pharmaceutical compositions are known in the art. Exemplary pharmaceutical compositions may be found, for example, in Tovey, 2018, Pharmaceutical Formulation: The Science and Technology of Dosage Forms: Volume 64 (Drug Discovery), Royal Society of Chemistry, and Niazi, 2019, Handbook of Pharmaceutical Manufacturing Formulations (3rdEd.), CRC Press (each herein incorporated by reference in their entirety).

[0450] In a related aspect, the invention provides the composition provided herein for use in medicine.

[0451] In a further related aspect, the invention provides the composition provided herein for use as a medicament.

[0452] In a further related aspect, the invention provides the composition provided herein for use in a method of treating cancer.In a further related aspect, the invention provides use of the composition provided herein in manufacturing a medicament for the treatment of cancer.

[0453] In one aspect, the invention provides a method of treating cancer using the composition provided herein.

[0454] In some embodiments, the medical and / or therapeutic uses and methods provided herein comprise administering an effective dose of the composition to a subject.

[0455] In some embodiments, the subject has cancer.

[0456] In some embodiments, the cancer is a tumour solid cancer. In preferred embodiments, the cancer is an immune-excluded solid tumour cancer and / or an immune-excluded tumour. As used herein, the terms "immune-excluded solid tumour cancer" and "immune-excluded tumour" are interchangeable. As discussed herein, immune-excluded (or "immunologically cold") tumours are characterised by a tumour microenvironment (TME) which physically excludes immune cells from the tumour core, thereby limiting the efficacy of cancer immunotherapies. Other characteristics of immune-excluded (or "immunologically cold") tumours include but are not limited to: high levels of stromal deposition which prevents effective infiltration of different subpopulations of immune cells into the tumour microenvironment; collapsed vasculature; low levels of oxygen; an acidic pH; production of glucose; production of N-acetylglucosamine; production of ethanolamine; production of cAMP; and production of lactate.

[0457] Cancers and tumours that may suitably be treated with the recombinant bacterial cell or cell provided herein are disclosed in Wilkinson et al. (2017) Oxford Handbook of Clinical Medicine (10th Ed), Oxford University Press, and Cassidy et al. (2015) Oxford Handbook of Oncology (4thEd), Oxford University Press (each herein incorporated by reference in their entirety). In some embodiments, the cancer is a cancer selected from the group comprising or consisting of: triple-negative breast cancer, microsatellite stable colon cancer, and pancreatic cancer. The cancer may be a primary cancer or a secondary cancer, for example a metastatic cancer.

[0458] In one aspect, the invention provides a method for degrading extracellular matrix (ECM) in a tissue, comprising contacting the tissue with the recombinant bacterial cell provided herein, the cell provided herein, or the composition provided herein.The method may be an in vitro method or an in vivo method. In some embodiments, the method is an in vitro method. In some embodiments the method is an in vivo method, for example an in vivo medical method. In some embodiments, the method comprises administering the recombinant bacterial cell, the cell, or the composition to a subject. The subject may be any subject discussed herein. In some embodiments, the subject has a disease. In preferred embodiments, the disease is cancer. The cancer may be any cancer as discussed herein; however, in some embodiments, the cancer comprises extracellular matrix. In some embodiments the cancer is a solid tumour cancer, optionally wherein the cancer is an immune-excluded solid cancer. In some embodiments, the cancer is a cancer selected from the group comprising or consisting of: triple-negative breast cancer, microsatellite stable colon cancer, and pancreatic cancer. The cancer may be a primary cancer or a secondary cancer, for example a metastatic cancer. In some embodiments, the cancer is not a cancer that does not comprise extracellular matrix. In some embodiments, the cancer is not a blood cancer. In some embodiments, the cancer is not a cancer selected from the group comprising or consisting of: leukaemia, myeloma, lymphoma.

[0459] As disclosed herein, the subject may be any suitable subject; however, in some embodiments, the subject is a human subject.

[0460] In one aspect, the invention provides use of the recombinant bacterial cell provided herein, the cell provided herein, or the composition provided herein to degrade extracellular matrix (ECM). In some embodiments, the use is an in vitro use or an in vivo use. In some embodiments, the use comprises administering the recombinant bacterial cell, the cell, or the composition to a subject. The subject may be any subject provided herein.

[0461] In embodiments comprising the administration of the recombinant bacterial cell, cell, or composition provided herein, in some embodiments the recombinant bacterial cell, the cell, or the composition is administered to a subject by a route selected from the group comprising or consisting of: oral administration, intraocular administration, intravenous administration, intraarterial administration, intraperitoneal administration, intramuscular administration, intratumoural administration, buccal administration, nasal administration, pulmonary administration. In some embodiments, the recombinant bacterial cell, the cell, or the composition is administered by injection, for example by intravenous injection, intratumoural injection, intraperitoneal injection, intraocular injection, intraarterial injection, or intramuscular injection. In some embodiments, the recombinant bacterial cell, the cell, or the composition is administered as a tablet, a capsule, a powder, a bolus, a suspension, or a solution, for example by oral administration. In some embodiments,the recombinant bacterial cell, the cell, or the composition is administered as a spray, for example by nasal administration and / or by pulmonary administration.

[0462] In some embodiments, following administration the recombinant bacterial cell or cell accumulates in a target tissue or tumour in the subject. The term "accumulating in a target tissue or tumour" or "accumulates in a target tissue or tumour" is discussed herein.

[0463] In some embodiments, the recombinant bacterial cell or cell degrades the extracellular matrix (ECM) of the target tissue or tumour. For example, the recombinant bacterial cell or cell may degrade any extracellular matrix (ECM) component disclosed herein, for example by expressing an extracellular matrix (ECM) degrading polypeptide as provided herein. The extracellular matrix (ECM) degrading polypeptide may be any extracellular matrix (ECM) degrading polypeptide provided herein; however, in a preferred embodiment the extracellular matrix (ECM) degrading polypeptide is secreted by the cell. Without wishing to be bound by theory, such degradation of extracellular matrix (ECM) in a target tissue or tumour will allow the entry of immune cells and / or cancer therapeutics such as immunotherapy agents into the tissue or tumour. Without wishing to be bound by theory, it is thought that entry of immune cells into immune excluded tumours will enable the efficacy of immunotherapeutic strategies which activate said immune cells to eliminate cancer cells. Entry of a cancer therapeutic into the tissue or tumour will allow the cancer therapeutic to have the relevant therapeutic effect on the tissue or tumour, or will improve the therapeutic effect of the cancer therapeutic on the tissue or tumour. In other words, when co-administered, the recombinant bacterial cell provided herein or cell provided herein and cancer therapeutic have a synergistic therapeutic effect on the target tissue, tumour, and / or cancer.

[0464] As will be appreciated, it may be advantageous to administer the recombinant bacterial cell provided herein, the cell provided herein, or composition provided herein to a subject at the same time or at approximately the same time as the subject is administered a therapeutic agent. The therapeutic agent may be any suitable therapeutic agent; however, the therapeutic agent is preferably a cancer therapeutic. Accordingly, in one embodiment, the recombinant bacterial cell provided herein, the cell provided herein, or composition provided herein is:

[0465] a) administered prior to subsequent administration of a cancer therapeutic; and / or b) co-administered with a cancer therapeutic.

[0466] The cancer therapeutic may be any suitable cancer therapeutic; however, in preferred embodiments the cancer therapeutic is a cancer immunotherapy. In some embodiments,the cancer therapeutic is selected from the group comprising or consisting of: an antibody (optionally selected from the group comprising or consisting of: an immune checkpoint inhibitor (ICI), T-cell engager, and a bispecific antibody or an antibody-drug conjugate), a hormonal therapy, a targeted therapy, an immunotherapy, a cell therapy (optionally selected from the group comprising or consisting of: a T-cell, a CAR-T cell, and a T4 cell), a chemotherapy agent (optionally selected from the group comprising or consisting of: FOLFOX, Gemcitabine, Paclitaxel, Cisplatin, Epirubicin and Irinotecan) an Immunomodulator (optionally selected from the group comprising or consisting of: a cytokine and a chemokine), a vaccine (optionally an mRNA vaccine), a virus (optionally an oncolytic virus), and a second live biotherapeutic (optionally selected from the group comprising or consisting of: an engineered live biotherapeutic and a recombinant bacterial cell). In some embodiments, the immune checkpoint inhibitor is selected from the group comprising or consisting of: an anti-PD-1 antibody and an anti-CTLA-4 antibody, an anti-LAG3 antibody, an anti-PDL1 antibody, and anti-TIGIT antibody, an anti-TIM3 antibody, an anti-NKG2A antibody, an anti-BTLA antibody, and anti-GITR antibody, an anti-ICOS antibody, an anti-VISTA antibody. In some embodiments the cell therapy is selected from the group comprising or consisting of: A tumour infiltrating lymphocyte (TIL), a T cell with an engineered T cell receptor (TCR), a CAR T cell, a Natural Killer cell. In some embodiments the chemotherapy is selected from the group comprising or consisting of an alkylating agent (optionally temozolomide and carboplatin), an anti-metabolite (optionally 5-FU or gemcitabine), an anti-tumour antibiotic (optionally doxorubicin), a topoisomerase inhibitor (optionally irinotecan), a mitotic inhibitor (optionally a taxane, optionally paclitaxel or docetaxel), a corticosteroid (optionally dexamethasone).

[0467] In one aspect, the invention provides a pharmaceutical composition comprising the recombinant bacterial cell provided herein or the cell provided herein, and a cancer therapeutic, for use in treating cancer. In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier, diluent, excipient, adjuvant, buffer, salt, or the like. Suitable pharmaceutical compositions are known in the art. Exemplary pharmaceutical compositions may be found, for example, in Tovey, 2018, Pharmaceutical Formulation: The Science and Technology of Dosage Forms: Volume 64 (Drug Discovery), Royal Society of Chemistry, and Niazi, 2019, Handbook of Pharmaceutical Manufacturing Formulations (3rdEd.), CRC Press (each herein incorporated by reference in their entirety).

[0468] In one aspect, the invention provides a combination comprising the recombinant bacterial cell provide herein or cell provided herein and a cancer therapeutic for use in treating cancer.In one aspect, the invention provides a combination comprising a cancer therapeutic and the recombinant bacterial cell provided herein or cell provided herein for use in treating cancer.

[0469] In one embodiment, the combination may be provided as a composition, optionally wherein the composition is a pharmaceutical composition as described herein.

[0470] In one embodiment:

[0471] a) the cancer is a cancer that comprises extracellular matrix; optionally wherein the cancer is selected from the group comprising or consisting of: triple-negative breast cancer, microsatellite stable colon cancer, and pancreatic cancer; and / or b) the cancer is not a cancer that does not comprise extracellular matrix; optionally wherein the cancer is not a blood cancer; optionally wherein the cancer is not a cancer selected from the group comprising or consisting of: leukaemia, myeloma, and lymphoma.

[0472] In one embodiment, the cancer therapeutic is a cancer immunotherapy agent, optionally wherein the cancer immunotherapy agent is selected from the group comprising or consisting of: an antibody (optionally selected from the group comprising or consisting of: an immune checkpoint inhibitor (ICI), T-cell engager, and a bispecific antibody or an antibody-drug conjugate), a cell therapy (optionally selected from the group comprising or consisting of: a T-cell, a CAR-T cell, and a T4 cell), a chemotherapy agent (optionally selected from the group comprising or consisting of: FOLFOX, Gemcitabine, Paclitaxel, Cisplatin, Epirubicin and Irinotecan) an Immunomodulator (optionally selected from the group comprising or consisting of: a cytokine and a chemokine), a vaccine (optionally an mRNA vaccine), a virus (optionally an oncolytic virus), and a second live biotherapeutic (optionally selected from the group comprising or consisting of: an engineered live biotherapeutic and a recombinant bacterial cell). In some embodiments, the immune checkpoint inhibitor is selected from the group comprising or consisting of: an anti-PD-1 antibody and an anti-CTLA-4 antibody, an anti-LAG3 antibody, an anti-PDL1 antibody, and anti-TIGIT antibody, an anti-TIM3 antibody, an anti-NKG2A antibody, an anti-BTLA antibody, and anti-GITR antibody, an anti-ICOS antibody, an anti-VISTA antibody. In some embodiments the cell therapy is selected from the group comprising or consisting of: A tumour infiltrating lymphocyte (TIL), a T cell with an engineered T cell receptor (TCR), a CAR T cell, a Natural Killer cell. In some embodiments the chemotherapy is selected from the group comprising or consisting of an alkylating agent (optionally temozolomide and carboplatin), an anti-metabolite (optionally 5-FU or gemcitabine), an anti-tumour antibiotic (optionally doxorubicin), a topoisomerase inhibitor (optionally irinotecan), a mitoticinhibitor (optionally a taxane, optionally paclitaxel or docetaxel), a corticosteroid (optionally dexamethasone).

[0473] In one embodiment, the composition or combination for use is administered to a subject. In one embodiment:

[0474] a) the recombinant bacterial cell or cell is administered prior to or subsequent to administration of the cancer therapeutic; and / or

[0475] b) the recombinant bacterial cell or cell is co-administered with the cancer therapeutic.

[0476] In one embodiment, the composition or combination for use is administered to a subject by a route selected from the group comprising or consisting of: oral administration, intraocular administration, intravenous administration, intraarterial administration, intraperitoneal administration, intramuscular administration, intratumoural administration, buccal administration, nasal administration, pulmonary administration. Suitable methods of administration via these routes are provided herein.

[0477] In one embodiment, following administration the recombinant bacterial cell or cell accumulates in a target tissue or tumour in the subject. The term "accumulating in a target tissue or tumour" or "accumulates in a target tissue or tumour" is discussed herein.

[0478] In one embodiment, the recombinant bacterial cell or cell degrades the extracellular matrix (ECM) of the target tissue or tumour. For example, the recombinant bacterial cell or cell may degrade any extracellular matrix (ECM) component disclosed herein, for example by expressing an extracellular matrix (ECM) degrading polypeptide as provided herein. The extracellular matrix (ECM) degrading polypeptide may be any extracellular matrix (ECM) degrading polypeptide provided herein; however, in a preferred embodiment the extracellular matrix (ECM) degrading polypeptide is secreted by the cell. Without wishing to be bound by theory, such degradation of extracellular matrix (ECM) in a target tissue or tumour will allow the entry of immune cells and / or cancer therapeutics such as immunotherapy agents into the tissue or tumour.

[0479] Accordingly, in on embodiment, degradation of the extracellular matrix (ECM) of the target tissue or tumour allows the host's immune cells or an administered cancer therapeutic to contact and / or enter the tumour. In one embodiment, said contacting and / or entering the tumour is required for or improves the therapeutic effect of the cancer therapeutic. In other words, when co-administered, the recombinant bacterial cell provided herein or cellprovided herein and cancer therapeutic have a synergistic therapeutic effect on the target tissue, tumour, and / or cancer.

[0480] As will be understood, recombinant bacterial cells and cells provided herein, when administered to a subject, do not persist in and / or do not colonise the subject prior to, during, or following localisation to a target tissue or tumour and / or degradation of extracellular matrix (ECM) in a target tissue or tumour. For example, the recombinant bacterial cells may lyse, be cleared by the immune system during and / or following localisation to a target tissue or tumour and / or degradation of extracellular matrix (ECM) in a target tissue or tumour, or will otherwise be exhausted during and / or following localisation to a target tissue or tumour and / or degradation of extracellular matrix (ECM) in the target tissue or tumour. In this way, the recombinant bacterial cell or cell is exhausted or "used up" during the course of its therapeutic and / or medical use, when administered alone or in combination with a cancer therapeutic, including when administered as part of a composition. As set out herein, in preferred embodiments the nucleic acids provided herein do not comprise an antibiotic resistance marker. Accordingly, in preferred embodiments the recombinant bacterial cell provided herein is not resistant to an antibiotic. A recombinant bacterial cell or cell provided herein may also be cleared from a subject by antibiotic treatment. Suitable antibiotic treatments are known to the person skilled in the art.

[0481] In one aspect, the invention provides a kit of parts, comprising:

[0482] a) the recombinant bacterial cell provided herein;

[0483] b) the engineered hypoxia-inducible promoter provided herein;

[0484] c) the engineered pH-inducible promoter provided herein;

[0485] d) the nucleic acid provided herein;

[0486] e) the cell provided herein;

[0487] f) the composition provided herein;

[0488] g) a therapeutic agent, optionally an anti-cancer therapeutic.

[0489] In one aspect, the invention provides a recombinant bacterial cell, a cell, a pH-inducible promoter, a hypoxia-inducible promoter, a nucleic acid, a use, a composition, a method, and a kit as described substantially herein.

[0490] The invention also provides the following numbered paragraphs embodiments:

[0491] 1. A recombinant bacterial cell capable of expressing a heterologous extracellular matrix (ECM) degrading polypeptide that degrades an extracellular matrix component, wherein:the cell comprises a nucleic acid comprising a nucleotide sequence that encodes the heterologous extracellular matrix (ECM) degrading polypeptide and a promoter;

[0492] wherein the nucleotide sequence that encodes the heterologous extracellular matrix (ECM) degrading polypeptide is operably linked to the promoter; and

[0493] the promoter is a tumour-inducible promoter.

[0494] 2. The recombinant bacterial cell of paragraph 1, wherein the heterologous extracellular matrix (ECM) degrading polypeptide is secreted from the cell.

[0495] 3. A recombinant bacterial cell capable of expressing a heterologous extracellular matrix (ECM) degrading polypeptide that degrades an extracellular matrix component, wherein the cell comprises a nucleic acid comprising a nucleotide sequence that encodes the heterologous extracellular matrix (ECM) degrading polypeptide,

[0496] wherein the heterologous extracellular (ECM) degrading polypeptide is secreted from the cell.

[0497] 4. The recombinant bacterial cell of paragraph 3, wherein the nucleic acid further comprises a promoter; optionally

[0498] wherein the nucleotide sequence encoding the heterologous extracellular matrix degrading (ECM) polypeptide is operably linked to the promoter and wherein the promoter is a tumour-inducible promoter.

[0499] 5. The recombinant bacterial cell of any one of paragraphs 1, 2, and 4 wherein the tumour-inducible promoter is:

[0500] a) a hypoxia-inducible promoter; optionally wherein the hypoxia-inducible promoter comprises:

[0501] i) an FNR binding motif;

[0502] ii) an fnrS promoter or subsequence thereof; and / or

[0503] iii) a grcA promoter or subsequence thereof;

[0504] b) an N-acetylglucosamine-inducible promoter; optionally wherein the N- acetylglucosamine-inducible promoter comprises a NagC binding motif;

[0505] c) an ethanolamine-inducible promoter; optionally wherein the ethanolamine- inducible promoter comprises a EutR binding motif;

[0506] d) a lactate-inducible promoter; optionally wherein the lactate-inducible promoter comprises an IldR binding motif;

[0507] e) a pH-inducible promoter;

[0508] f) a cAMP-inducible promoter; optionally wherein the cAMP-inducible promoter comprises a CRP binding motif; optionallyg) a hypoxia- and N-acetylglucosamine-inducible promoter; optionally wherein the hypoxia- and N-acetylglucosamine-inducible promoter comprises a NagC binding motif and:

[0509] i) an FNR binding motif;

[0510] ii) an fnrS promoter or subsequence thereof; and / or

[0511] iii) a grcA promoter or subsequence thereof;

[0512] h) a hypoxia- and ethanolamine-inducible promoter; optionally wherein the hypoxia- and ethanolamine-inducible promoter comprises a EutR binding motif and:

[0513] i) an FNR binding motif;

[0514] ii) an fnrS promoter or subsequence thereof; and / or

[0515] iii) a grcA promoter or subsequence thereof;

[0516] i) a hypoxia- and lactate-inducible promoter; optionally wherein the hypoxia-and lactate-inducible promoter comprises an IldR binding motif and:

[0517] i) an FNR binding motif;

[0518] ii) an fnrS promoter or subsequence thereof; and / or

[0519] iii) a grcA promoter or subsequence thereof;

[0520] j) a hypoxia- and pH-inducible promoter; optionally

[0521] wherein the hypoxia- and pH-inducible promoter comprises:

[0522] i) an FNR binding motif; and / or

[0523] ii) an fnrS promoter or subsequence thereof; and / or

[0524] iii) a grcA promoter or subsequence thereof;

[0525] k) a hypoxia- and cAMP-inducible promoter; optionally wherein the hypoxia-and cAMP-inducible promoter comprises a CRP binding motif and:

[0526] i) an FNR binding motif; and / or

[0527] ii) an fnrS promoter or subsequence thereof; and / or

[0528] iii) a grcA promoter or subsequence thereof;

[0529] l) a pH- and lactate-inducible promoter; optionally wherein the pH- and lactate-inducible promoter comprises an IldR binding motif;

[0530] m) a hypoxia-, N-acetylglucosamine, and lactate-inducible promoter; optionally wherein the hypoxia-, N-acetylglucosamine, and lactate-inducible promoter comprises a NagC binding motif, an IldR binding motif, and:

[0531] i) an FNR binding motif; and / or

[0532] ii) an fnrS promoter or subsequence thereof; and / or

[0533] iii) a grcA promoter or subsequence thereof;

[0534] n) a hypoxia-, cAMP, and lactate-inducible promoter; optionally wherein the hypoxia-, cAMP, and lactate-inducible promoter comprises a CRP binding motif, an IldR binding motif, and:

[0535] i) an FNR binding motif; and / orii) an fnrS promoter or subsequence thereof; and / or

[0536] iii) a grcA promoter or subsequence thereof;

[0537] o) an ethanolamine- and lactate-inducible promoter; optionally wherein the ethanolamine- and lactate-inducible promoter comprises a EutR binding motif and an IldR binding motif;

[0538] p) an ethanolamine-, lactate-, and N-acetylglucosamine-inducible promoter; optionally wherein the ethanolamine-, lactate-, and N-acetylglucosamine-inducible promoter comprises a EutR binding motif, an IldR binding motif, and a NagC binding motif;

[0539] q) an ethanolamine-, lactate-, and hypoxia-inducible promoter; optionally wherein the ethanolamine-, lactate-, and hypoxia-inducible promoter comprises a EutR binding motif, an IldR binding motif, and:

[0540] i) an FNR binding motif; and / or

[0541] ii) an fnrS promoter or subsequence thereof; and / or

[0542] iii) a grcA promoter or subsequence thereof;

[0543] and / or

[0544] r) an ethanolamine-, lactate-, hypoxia-, and N-acetylglucosamine-inducible promoter; optionally wherein the ethanolamine-, lactate-, hypoxia- and N- acetylglucosamine-inducible promoter comprises a EutR binding motif, an IldR binding motif, a NagC binding motif and:

[0545] i) an FNR binding motif; and / or

[0546] ii) an fnrS promoter or subsequence thereof; and / or

[0547] iii) a grcA promoter or subsequence thereof;

[0548] 6. The tumour-inducible promoter of paragraph 5, wherein:

[0549] a) the FNR binding motif comprises or consists of a nucleotide sequence of: SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244;

[0550] b) the fnrS promoter or subsequence thereof comprises or consists of a nucleotide sequence of: SEQ ID NO: 250 or SEQ ID NO: 257, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 250 or SEQ ID NO: 257;c) the NagC binding motif comprises or consists of a nucleotide sequence of: SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 239, or SEQ ID NO: 259, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 239, or SEQ ID NO: 259;

[0551] d) the EutR binding motif comprises or consists of a nucleotide sequence of: SEQ ID NO: 261 or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 261;

[0552] e) the IldR binding motif comprises or consists of a nucleotide sequence of: SEQ ID NO: 248, SEQ ID NO: 249, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249;

[0553] f) the grcA promoter or subsequence thereof comprises or consists of a nucleotide sequence of: SEQ ID NO: 253 or SEQ ID NO: 254, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249;

[0554] and / or

[0555] g) CRP binding motif comprises or consists of a nucleotide sequence of: SEQ ID NO: 245 or SEQ ID NO: 246; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 245 or SEQ ID NO: 246.

[0556] 7. The recombinant bacterial cell of any one of paragraphs 1, 2, and 4-6, wherein the tumour-inducible promoter comprises:

[0557] a) a discriminator; optionally

[0558] wherein the discriminator comprises or consists of a nucleotide sequence of: SEQ ID NO: 229, SEQ ID NO: 230, SEQ ID NO: 231, SEQ ID NO: 232, SEQ ID NO: 233, SEQ ID NO: 234, SEQ ID NO: 235, SEQ ID NO: 236, SEQ ID NO: 237, SEQ ID NO: 238, SEQ ID NO: 240, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 229, SEQ ID NO: 230, SEQ ID NO: 231, SEQ ID NO: 232, SEQ ID NO: 233, SEQ ID NO: 234,SEQ ID NO: 235, SEQ ID NO: 236, SEQ ID NO: 237, SEQ ID NO: 238, or SEQ ID NO: 240; optionally

[0559] wherein the discriminator is a "high GC" discriminator; optionally wherein the "high GC" discriminator has a sequence of SEQ ID NO: 232 or SEQ ID NO: 233;

[0560] b) a -10 / -35 spacer; optionally

[0561] wherein the 10 / -35 spacer is selected from the group comprising or consisting of: SEQ ID NO: 217, SEQ ID NO: 218, SEQ ID NO: 219, SEQ ID NO: 220, SEQ ID NO 221, SEQ ID NO: 222, SEQ ID NO: 223, SEQ ID NO: 224, SEQ ID NO: 225, or SEQ ID NO: 226, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 217, SEQ ID NO: 218, SEQ ID NO: 219, SEQ ID NO: 220, SEQ ID NO 221, SEQ ID NO: 222, SEQ ID NO: 223, SEQ ID NO: 224, SEQ ID NO: 225, or SEQ ID NO: 226;

[0562] c) a TUSP binding site; optionally

[0563] wherein the TUSP binding site comprises or consists of a nucleotide sequence of: SEQ ID NO: 227, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 227;

[0564] d) a TGIF sequence; optionally

[0565] wherein the TGIF sequence comprises or consists of a nucleotide sequence of: SEQ ID NO: 230, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 230; or

[0566] e) a terminator sequence; optionally

[0567] wherein the terminator sequence comprises or consists of a nucleotide sequence of: SEQ ID NO: 247, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 247;

[0568] f) a ribosome binding site (RBS); optionally

[0569] wherein the ribosome binding site (RBS) comprises or consists of a nucleotide sequence of: SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ IDNO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, or SEQ ID NO: 105; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100. SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, or SEQ ID NO: 105.

[0570] 8. The recombinant bacterial cell of any one of paragraphs 1, 2, and 4-7, wherein the tumour-inducible promoter comprises or consists of a nucleotide sequence of:

[0571] a) SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO; 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO; 135, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO; 145, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO; 155, SEQ ID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO; 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO; 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO: 179, SEQ ID NO: 180, SEQ ID NO: 181, SEQ ID NO: 182, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO; 185, SEQ ID NO: 186, SEQ ID NO: 187, SEQ ID NO: 188, SEQ ID NO: 189, SEQ ID NO: 190, SEQ ID NO: 191, SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO; 195, SEQ ID NO: 196, SEQ ID NO: 197, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO; 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210, SEQ ID NO: 275, SEQ ID NO: 276, SEEQ ID NO: 277, or SEQ ID NO: 278, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ IDNO; 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO; 135, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO; 145, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO; 155, SEQ ID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO; 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO; 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO: 179, SEQ ID NO: 180, SEQ ID NO: 181, SEQ ID NO: 182, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO; 185, SEQ ID NO: 186, SEQ ID NO: 187, SEQ ID NO: 188, SEQ ID NO: 189, SEQ ID NO: 190, SEQ ID NO: 191, SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO; 195, SEQ ID NO: 196, SEQ ID NO: 197, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO; 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210, SEQ ID \IO: 275, SEQ ID I O: 276, SEEQ ID I 10: 277, or SEQ II > NO: 278; and / or b) SEQ ID NO: 160, SEQ ID NO: 182, SEQ ID NO: 190, SEQ ID NO: 128, SEQ ID NO: 125, SEQ ID NO: 131, SEQ ID NOL 132, SEQ ID NO: 133, SEQ ID NO: 144, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 185: SEQ ID NO: 204, SEQ ID NO: 209; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 160, SEQ ID NO: 182, SEQ ID NO: 190, SEQ ID NO: 128, SEQ ID NO: 125, SEQ ID NO: 131, SEQ ID NOL 132, SEQ ID NO: 133, SEQ ID NO: 144, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 185: SEQ ID NO: 204, SEQ ID NO: 209; and / or

[0572] c) SEQ ID NO: 160, SEQ ID NO: 182, SEQ ID NO: 190, SEQ ID NO: 128, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 160, SEQ ID NO: 182, SEQ ID NO: 190, SEQ ID NO: 128;

[0573] d) SEQ ID NO: 160; or

[0574] e) SEQ ID NO: 182; or

[0575] f) SEQ ID NO: 190; or

[0576] g) SEQ ID NO: 128; or

[0577] h) SEQ ID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278; and / or

[0578] i) SEQ ID NO: 275; orj) SEQ ID NO: 276; or

[0579] k) SEQ ID NO: 277; or

[0580] l) SEQ ID NO: 278; or

[0581] m) SEQ ID NO: 125; or

[0582] n) SEQ ID NO: 131; or

[0583] o) SEQ ID NO: 132; or

[0584] p) SEQ ID NO: 133; or

[0585] q) SEQ ID NO: 144; or

[0586] r) SEQ ID NO: 170; or

[0587] s) SEQ ID NO: 171; or

[0588] t) SEQ ID NO: 185; or

[0589] u) SEQ ID NO: 204; or

[0590] v) SEQ ID NO: 209.

[0591] 9. The recombinant bacterial cell of any one of paragraphs 1, 2, and 4-8s, wherein the tumour-inducible promoter is:

[0592] a) a hypoxia-inducible promoter, optionally wherein the hypoxia-inducible promoter comprises or consists of a nucleotide sequence of:

[0593] i) SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ D NO: 132, SEQ UD NO: 133, SEQ ID NO 135, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO; 155, SEQ ID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO; 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO; 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO: 179, SEQ ID NO: 180, SEQ ID NO: 189, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO; 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210, SEQ ID NO: 275, or SEQ ID NO: 278; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ IDNO: 54, SEQ ID NO: 55, or SEQ ID NO: 56, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ D NO: 132, SEQ UD NO: 133, SEQ ID NO 135, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: 150, SEQ ID NO: 179, SEQ ID NO: 180, SEQ ID NO: 189, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO; 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, or SEQ ID NO: 210, SEQ ID NO: 275, or SEQ ID NO: 278; and / or

[0594] ii) SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ D NO: 132, SEQ UD NO: 133, SEQ ID NO 135, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: SEQ ID NO: 150, SEQ ID NO: 179, or SEQ ID NO: 180; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ D NO: 132, SEQ UD NO: 133, SEQ ID NO 135, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: SEQ ID NO: 150, SEQ ID NO: 179, or SEQ ID NO: 180; and / or

[0595] iii) SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 128, or SEQ ID NO: 278, SEQ ID NO: 125, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO: 144, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 204, or SEQ ID NO: 209; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 128, or SEQ ID NO: 278, SEQ ID NO: 125, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO: 144, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 204, or SEQ ID NO: 209; and / or

[0596] iv) SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 128, or SEQ ID NO: 278; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%,at least 98%, or at least 99% identical to SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 128, or SEQ ID NO: 278; and / or

[0597] v) SEQ ID NO: 160; or

[0598] vi) SEQ ID NO: 275; or

[0599] vii) SEQ ID NO: 128; or

[0600] viii) SEQ ID NO: 278; or

[0601] ix) SEQ ID NO: 125; or

[0602] x) SEQ ID NO: 131; or

[0603] xi) SEQ ID NO: 132; or

[0604] x) SEQ ID NO: 133; or

[0605] xi) SEQ ID NO: 144; or

[0606] xii) SEQ ID NO: 170; or

[0607] xiii) SEQ ID NO: 171; or

[0608] xiv) SEQ ID NO: 204; or

[0609] xv) SEQ ID NO: 209;

[0610] b) an N-acetylglucosamine-inducible promoter; optionally wherein the N-acetylglucosamine-inducible promoter comprises or consists of a nucleotide sequence of:

[0611] i) the N-acetylglucosamine-inducible promoter comprises a sequence of SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 169, SEQ ID NO: 172, SEQ ID NI: 173, SEQ ID NO: 194, SEQ ID NO: 196, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, or SEQ ID NO: 207; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 169, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 194, SEQ ID NO: 196, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, or SEQ ID NO: 207; and / or

[0612] ii) SEQ ID NO: 144 or SEQ ID NO: 204; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 144 or SEQ ID NO: 204; and / or

[0613] iii) SEQ ID NO: 144; or

[0614] iv) SEQ ID NO: 204;

[0615] c) an ethanolamine-inducible promoter; optionally wherein the ethanolamine-inducible promoter comprises or consists of a nucleotide sequence of:i) SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, or SEQ ID NO: 197; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, or SEQ ID NO: 197; and / or

[0616] ii) SEQ ID NO: 192 or SEQ ID NO: 197; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 192 or SEQ ID NO: 197;

[0617] d) a lactate-inducible promoter; optionally wherein the lactate-inducible promoter comprises or consists of a nucleotide sequence of:

[0618] i) SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO: 155, SEQ ID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO; 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO; 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO; 189, SEQ ID NO: 190, SEQ ID NO: 191, SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210, SEQ ID NO: 275, or SEQ ID NO: 277; or a sequence is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO: 155, SEQ ID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO; 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO; 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO; 189, SEQ ID NO: 190, SEQ ID NO: 191, SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210, SEQ ID NO: 275, or SEQ ID NO: 277; and / or

[0619] ii) SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 190, SEQ ID NO: 277, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 204, or SEQ ID NO: 209; or anucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 190, SEQ ID NO: 277, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 204, or SEQ ID NO: 209;

[0620] iii) SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 190, or SEQ ID NO: 277; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 190, or SEQ ID NO: 277; and / or

[0621] iv) SEQ ID NO: 160; or

[0622] iv) SEQ ID NO: 275; or

[0623] v) SEQ ID NO: 190; or

[0624] vi) SEQ ID NO: 277; or

[0625] vii) SEQ ID NO: 170; or

[0626] viii) SEQ ID NO: 171; or

[0627] ix) SEQ ID NO: 204; or

[0628] x) SEQ ID NO: 209;

[0629] e) a pH-inducible promoter; optionally wherein the pH-inducible promoter comprises or consists of a nucleotide sequence of:

[0630] i) SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO 181, SEQ ID NO: 182, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 187, SEQ ID NO: 188, SEQ ID NO: 190, SEQ ID NO: 191, SEQ ID NO: 276, or SEQ ID NO: 277; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, or SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO 181, SEQ ID NO: 182, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 187, SEQ ID NO: 188, SEQ ID NO: 190, SEQ ID NO: 191, SEQ ID NO: 276, or SEQ ID NO: 277; and / or

[0631] ii) SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO 181, SEQ ID NO: 182, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 187, SEQ ID NO: 276, or SEQ ID NO: 277; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%,or at least 99% identical to SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO 181, SEQ ID NO: 182, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 187, SEQ ID NO: 276, or SEQ ID NO: 277; and / or

[0632] iii) SEQ ID NO: 182, SEQ ID NO: 276, SEQ ID NO: 190, or SEQ ID NO: 277; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 182, SEQ ID NO: 276, SEQ ID NO: 190, or SEQ ID NO: 277; and / or

[0633] iv) SEQ ID NO: 182; or

[0634] v) SEQ ID NO: 276; or

[0635] vi) SEQ ID NO: 190; or

[0636] vii) SEQ ID NO: 277; or

[0637] viii) SEQ ID NO: 185;

[0638] f) a cAMP-inducible promoter; optionally wherein the cAMP-inducible promoter comprises or consists of a nucleotide sequence of:

[0639] i) SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 170, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 189, SEQ ID NO: 208, SEQ ID NO: 209, or SEQ ID NO: 210; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to: SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 170, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 189, SEQ ID NO: 208, SEQ ID NO: 209, or SEQ ID NO: 210; and / or

[0640] ii) SEQ ID NO: 170 or SEQ ID NO: 209; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to: SEQ ID NO: 170 or SEQ ID NO: 209; and / or

[0641] iii) SEQ ID NO: 170; or

[0642] iv) SEQ ID NO: 209;

[0643] g) a hypoxia- and N-acetylglucosamine-inducible promoter; optionally wherein the hypoxia- and N-acetylglucosamine-inducible promoter comprises or consists of a nucleotide sequence of:

[0644] i) SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 169, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 196, SEQ ID NO: 198, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 169, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 196, SEQ ID NO: 198, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207; and / or

[0645] ii) SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 198, SEQ ID NO: 200, SEQ ID NO: 204; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 198, or SEQ ID NO: 200, SEQ ID NO: 204; or

[0646] iii) SEQ ID NO: 144; or

[0647] iv) SEQ ID NO: 204;

[0648] h) a hypoxia- and ethanolamine-inducible promoter; optionally wherein:

[0649] the hypoxia- and ethanolamine-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 195 or SEQ ID NO: 196; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 195 or SEQ ID NO: 196;

[0650] i) a hypoxia- and lactate-inducible promoter; optionally wherein the hypoxia-and lactate-inducible promoter comprises or consists of a nucleotide sequence of:

[0651] i) SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 156, SEQ ID NO: 157, SEQ IDNO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO: 189, SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207, or SEQ ID NO: 275; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 156, SEQ ID NO: 157, SEQ IDNO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176, SEQ IDNO: 177, SEQ ID NO: 178, SEQ ID NO: 189, SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207, or SEQ ID NO: 275; and / or

[0652] ii) SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 171, SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 178, or SEQ ID NO: 275; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 156, SEQ ID NO: 157, SEQ IDNO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 171, SEQ ID NO: 174, SEQ ID NO: 175 SEQ ID NO: 178, or SEQ ID NO: 275; and / or

[0653] iii) SEQ ID NO: 160 or SEQ ID NO: 275; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 160 or SEQ ID NO: 275; and / or

[0654] iv) SEQ ID NO: 160; or

[0655] v) SEQ ID NO: 275; or

[0656] vi) SEQ ID NO: 170; or

[0657] vii) SEQ ID NO: 171; or

[0658] viii) SEQ ID NO: 204; or

[0659] ix) SEQ ID NO: 209;

[0660] j) a hypoxia- and pH-inducible promoter; optionally wherein the pH-inducible and hypoxia-inducible promoter comprises or consists of a nucleotide sequence of:

[0661] SEQ ID NO: 73 or SEQ ID NO: 188; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 73 or SEQ ID NO: SEQ ID NO: 188;

[0662] k) a hypoxia- and cAMP-inducible promoter; optionally wherein the hypoxia-and cAMP-inducible promoter comprises or consists of a nucleotide sequence of:

[0663] i) SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 170, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 189, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%,at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 170, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 189, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210; and / or

[0664] ii) SEQ ID NO: 148, SEQ ID NO: 149; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 148, or SEQ ID NO: 149; and / or

[0665] iii) SEQ ID NO: 170, or SEQ ID NO: 209; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 170, or SEQ ID NO: 209; and / or

[0666] iv) SEQ ID NO: 148; or

[0667] v) SEQ ID NO: 149; or

[0668] vi) SEQ ID NO: 170; or

[0669] vii) SEQ ID NO: 209. or

[0670] l) a pH- and lactate-inducible promoter; optionally wherein the pH- and lactate-inducible promoter comprises or consists of a nucleotide sequence of:

[0671] i) SEQ ID NO: 190, SEQ ID NO: 191, or SEQ ID NO: 277; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 190, SEQ ID NO: 191, or SEQ ID NO: 277; and / or

[0672] ii) SEQ ID NO: 190 or SEQ ID NO: 277; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 190 or SEQ ID NO: 277; and / or

[0673] iii) SEQ ID NO: 190; or

[0674] iv) SEQ ID NO: 277;

[0675] m) a hypoxia-, N-acetylglucosamine, and lactate-inducible promoter; optionally wherein the hypoxia-, N-acetylglucosamine, and lactate-inducible promoter comprises or consists of a nucleotide sequence of:

[0676] i) SEQ ID NO: 169, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 205, SEQ ID NO: 206, or SEQ ID NO: 206; or sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 169, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: SEQ ID NO: 201, SEQ IDNO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 205, SEQ ID NO: 206, or SEQ ID NO: 206; and / or

[0677] ii) SEQ ID NO: 204;

[0678] n) a hypoxia-, cAMP, and lactate-inducible promoter; optionally wherein the hypoxia-, cAMP, and lactate-inducible promoter comprises or consists of a nucleotide sequence of:

[0679] i) SEQ ID NO: 170, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 189, SEQ ID NO: 208, SEQ ID NO: 209, or SEQ ID NO: 210; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 170, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 189, SEQ ID NO: 208, SEQ ID NO: 209, or SEQ ID NO: 210; and / or

[0680] ii) SEQ ID NO: 170; or

[0681] iii) SEQ ID NO: 209;

[0682] o) an ethanolamine- and lactate-inducible promoter; optionally wherein the ethanolamine- and lactate-inducible promoter comprises or consists of a nucleotide sequence of:

[0683] i) SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, or SEQ ID NO: 196; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, or SEQ ID NO: 196; and / or

[0684] ii) SEQ ID NO: 193 or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 193;

[0685] p) an ethanolamine-, lactate-, and N-acetylglucosamine-inducible promoter; optionally wherein the ethanolamine-, lactate-, and N-acetylglucosamine-inducible promoter comprises or consists of a nucleotide sequence of:

[0686] i) SEQ ID NO: 194 or SEQ ID NO: 196; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 194 or SEQ ID NO: 196; and / or

[0687] ii) SEQ ID NO: 194; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 194;

[0688] q) the tumour-inducible promoter is an ethanolamine-, lactate-, and hypoxiainducible promoter; optionally wherein the ethanolamine-, lactate-, and hypoxiainducible promoter comprises or consists of a nucleotide sequence of:i) SEQ ID NO: 195 or SEQ ID NO: 196; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 195 or SEQ ID NO: 196; and / or

[0689] ii) SEQ ID NO: 195; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 195; and / or

[0690] r) an ethanolamine-, lactate-, N-acetylglucosamine, and hypoxia-inducible promoter; optionally wherein the ethanolamine-, lactate-, N-acetylglucosamine, and hypoxia-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 196; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 196.

[0691] 9a. The recombinant bacterial cell of any one of paragraphs 5-9, wherein the hypoxiainducible promoter is not activated and does not drive expression of the heterologous ECM degrading polypeptide in nonmedia; optionally

[0692] wherein the hypoxia-inducible promoter is not activated and does not drive expression of the heterologous ECM degrading polypeptide at an oxygen concentration of:

[0693] a) above about 6%, above about 5%, above about 4%, or above about 3%; b) above 6%, above 5%, above 4%, or above 3%;

[0694] c) about 6%, about 5%, about 4%, about 3%; and / or

[0695] d) 6%, 5%, 4%, 3%.

[0696] 9b. The recombinant bacterial cell of any of paragraphs 5-9a, wherein the hypoxiainducible promoter is activated and drives expression of the heterologous ECM degrading polypeptide when the recombinant cell is incubated in culture media under mineral oil, optionally incubated at 37°C for 18 hours.

[0697] 9c. The recombinant bacterial cell of paragraph 9b, wherein the hypoxia-inducible promoter is activated and drives expression of the heterologous ECM degrading polypeptide at an oxygen concentration of:

[0698] a) between about 0% and about 2%, about 0.2% and about 1.8%, about 0.4% and about 1.6%, about 0.6% and about 1.4%, about 0.8% and about 1.2%, about 0% and about 1%, about 0.2% and about 0.8%, about 0.4 and about 0.6%, 1% and 2%, about 1.2% and about 1.8%, about 1.4% and about 1.6%;b) between 0% and 2%, 0.2% and 1.8%, 0.4% and 1.6%, 0.6% and 1.4%, 0.8% and 1.2%, 0% and 1%, 0.2% and 0.8%, 0.4 and 0.6%, 1% and 2%, 1.2% and 1.8%, 1.4% and 1.6%;

[0699] c) below about 2%, below about 1.5%, below about 1.6%, below about 1.4%, below about 1.2%, below about 1%, below about 0.8%, below about 0.6%, below about 0.4%, below about 0.2% or lower;

[0700] d) below 2%, below 1.5%, below 1.6%, below 1.4%, below 1.2%, below 1%, below 0.8%, below 0.6%, below 0.4%, below 0.2% or lower;

[0701] e) about 2%, about 1.8%, about 1.6%, about 1.4%, about 1.2%, about 1%, about 0.8%, about 0.6%, about 0.4%, about 0.2%; and / or

[0702] f) 2%, 1.8%, 1.6%, 1.4%, 1.2%, 1%, 0.8%, 0.6%, 0.4%, 0.2%.

[0703] 9d. The recombinant bacterial promoter of any one of paragraphs 5 to 9c, wherein the pH-inducible promoter is activated and drives expression of the heterologous ECM degrading polypeptide at an acidic pH; optionally

[0704] wherein the acidic pH is:

[0705] a) between about pH 5.5 and about pH 7, about pH 5.6 and about pH 6.9, about pH 5.7 and about pH 6.8, about pH 5.8 and about pH 6.7, about pH 5.9 and about pH 6.6, about pH 6.0 and about pH 6.5, about pH 6.1 and about pH 6.4, about pH 6.2 and about pH 6.3;

[0706] b) between pH 5.5 and pH 7, pH 5.6 and pH 6.9, pH 5.7 and pH 6.8, pH 5.8 and pH 6.7, pH 5.9 and pH 6.6, pH 6.0 and pH 6.5, pH 6.1 and pH 6.4, pH 6.2 and pH 6.3

[0707] c) below about pH 7, below about pH 6.9, below about pH 6.8, below about pH 6.7, below about pH 6.6, below about pH 6.5, below about pH 6.4, below about pH 6.3, below about pH 6.2, below about pH 6.1, below about pH 6.0, below about pH 5.9, below about pH 5.8, below about pH 5.7, below about pH 5.6, or lower; d) below pH 7, below pH 6.9, below pH 6.8, below pH 6.7, below pH 6.6, below pH 6.5, below pH 6.4, below pH 6.3, below pH 6.2, below pH 6.1, below pH 6.0, below pH 5.9, below pH 5.8, below pH 5.7, below pH 5.6;

[0708] e) about pH 5.5, about pH 5.6, about pH 5.7, about pH 5.8, about pH 5.9, about pH 6.0, about pH 6.1, about pH 6.2, about pH 6.3, about pH6.4, about pH 6.5, about pH 6.6, about pH 6.7, about pH 6.8, about pH 6.9, or about pH 7; and / or f) pH 5.5, pH 5.6, pH 5.7, pH 5.8, pH 5.9, pH 6.0, pH 6.1, pH 6.2, pH 6.3, pH6.4, pH 6.5, pH 6.6, pH 6.7, pH 6.8, pH 6.9, or pH 7.9e. The recombinant bacterial promoter of any one of paragraphs 5-9d, wherein the pH-inducible promoter is activated and drives expression of the heterologous ECM degrading polypeptide at an acidic pH; optionally

[0709] wherein the acidic pH is:

[0710] a) between about pH 5.9 and about pH 6.5, about pH 5.95 and about pH 6.45, about pH 6.0 and about pH 6.4, about pH 6.05 and about pH 6.35, about pH 6.1 and about 6.3, or about pH 6.15 and about pH 6.25;

[0711] b) between pH 5.9 and pH 6.5, pH 5.95 and pH 6.45, pH 6.0 and pH 6.4, pH 6.05 and pH 6.35, pH 6.1 and 6.3, or pH 6.15 and pH 6.25;

[0712] c) about pH 5.9, about pH 5.95, about pH 6.0, about pH 6.05, about pH 6.1, about pH 6.15, about pH 6.2, about pH 6.25, about pH 6.3, about pH 6.35, about pH 6.4, about pH 6.45, or about pH 6.5; and / or

[0713] d) pH 5.9, pH 5.95, pH 6.0, pH 6.05, pH 6.1, pH 6.15, pH 6.2, pH 6.25, pH 6.3, pH 6.35, pH 6.4, pH 6.45, or pH 6.5.

[0714] 9f. The recombinant bacterial cell of any of paragraphs 5-9e, wherein the pH-inducible promoter is not activated and does not drive expression of the heterologous ECM degrading polypeptide at an alkaline pH.

[0715] 9g. The recombinant bacterial cell of any one of paragraphs l-9f, wherein the tumourinducible promoter does not comprise or consist of a sequence of SEQ ID NO: 271, SEQ ID NO: 272, SEQ ID NO: 273, SEQ ID NO: 274.

[0716] 10. The recombinant bacterial cell of any one of paragraphs l-9g, wherein:

[0717] i) the extracellular matrix degrading polypeptide is:

[0718] a) a hyaluronidase;

[0719] b) a microbial hyaluronidase or a mammalian hyaluronidase;

[0720] c) a bacterial hyaluronidase;

[0721] d) chondroitin ABC lyase;

[0722] e) a microbial chondroitin ABC lyase or a mammalian chondroitin ABC lyase;

[0723] e) a bacterial chondroitin ABC lyase;

[0724] f) neuraminidase;

[0725] g) a microbial neuraminidase or a mammalian neuraminidase;

[0726] h) a bacterial neuraminidase;

[0727] i) PNGase;

[0728] j) a microbial PNGase or a mammalian PNGase;

[0729] k) a bacterial PNGase;optionally wherein the extracellular matrix component is a proteoglycan; ii) the ECM component is hyaluronic acid (HA) and the ECM degrading polypeptide is:

[0730] a) a hyaluronidase;

[0731] b) a microbial hyaluronidase or a mammalian hyaluronidase; and / or c) a bacterial hyaluronidase;

[0732] iii) the ECM component is Versican and the ECM degrading polypeptide is: a) chondroitin ABC lyase;

[0733] b) a microbial chondroitin ABC lyase or a mammalian chondroitin ABC lyase; and / or

[0734] c) a bacterial chondroitin ABC lyase;

[0735] iv) the ECM component is Versican and the ECM degrading polypeptide is: a) neuraminidase;

[0736] b) a microbial neuraminidase or a mammalian neuraminidase; and / or c) a bacterial neuraminidase;

[0737] v) the ECM component is Versican and the ECM degrading polypeptide is: a) PNGase;

[0738] b) a microbial PNGase or a mammalian PNGase; and / or

[0739] c) a bacterial PNGase;

[0740] vi) the heterologous extracellular matrix (ECM) degrading polypeptide:

[0741] a) is a hyaluronidase and comprises or consists of an amino acid sequence of SEQ ID NO: 1; is a chondroitin ABC Lyase and comprises or consists of an amino acid sequence of SEQ ID NO: 3 or 47; is a neuraminidase and comprises or consists of an amino acid sequence of SEQ ID NO: 5; or is a PNGase and comprises or consists of an amino acid sequence of SEQ ID NO: 7; or comprises or consists of an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, or SEQ ID NO: 47; and / or

[0742] b) is encoded by a nucleotide sequence of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, or SEQ ID NO: 48 or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, or SEQ ID NO: 48;

[0743] vii) the heterologous extracellular matrix (ECM) degrading polypeptide is a bacterial hyaluronidase; and / orviii) the heterologous extracellular matrix (ECM) degrading polypeptide is a bacterial hyaluronidase that:

[0744] a) comprises or consists of an amino acid sequence of SEQ ID NO: 1; or of an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 1; and / or b) is encoded by a nucleotide sequence of SEQ ID NO: 2, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 2.

[0745] 10a. The recombinant bacterial cell of any one of paragraphs 1-10, wherein the extracellular matrix (ECM) degrading polypeptide is not:

[0746] a) a hymenopteran hyaluronidase; optionally a bee venom hyaluronidase, a wasp venom hyaluronidase, or a tiger wasp venom hyaluronidase;

[0747] b) a human hyaluronidase; optionally human hyaluronidase-1 (hHyal-1), human hyaluronidase-2 (hHyal-2), human PH-20 (hPH-20), or bovine testes hyaluronidase (BTS); c) a Streptomyces koganeiensis hyaluronidase, a Streptomyces zooepidemic hyaluronidase, or a Streptomyces pristinaespiralis hyaluronidase;

[0748] d) a Streptococcus zooepidemicus hyaluronidase; or

[0749] e) a Penicillium funiculosum hyaluronidase;

[0750] f) an annelid hyaluronidase; optionally a leech hyaluronidase.

[0751] 11. The recombinant bacterial cell of any one of paragraphs l-10a, wherein the heterologous extracellular matrix (ECM) degrading polypeptide is a fusion polypeptide comprising an ECM degrading domain and:

[0752] a) a secretion domain;

[0753] b) two secretion domains;

[0754] c) a cleavage domain; and / or

[0755] d) a cleavage domain that is positioned between the ECM degrading domain and the secretion domain.

[0756] 12. The recombinant bacterial cell of paragraph 11, wherein:

[0757] i) the secretion domain is selected from the group comprising or consisting of: a PelB secretion signal sequence, a YebF carrier protein, a CtxB signal sequence, an AIDA-I autotransporter, a FliC signal sequence, and a Usp45 signal sequence, or any combination thereof;

[0758] ii) the secretion domain:a) comprises or consists of an amino acid sequence of SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35; or of an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35; and / or b) is encoded by a nucleotide sequence of SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36;

[0759] iii) the cleavage domain comprises a matrix metalloprotease (MMP) cleavage site, optionally wherein the cleavage domain:

[0760] a) comprises or consists of an amino acid sequence of SEQ ID NO: 116; or of an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 116; and / or b) is encoded by a nucleotide sequence of SEQ ID NO: 117, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 117; and / or

[0761] iv) the fusion polypeptide:

[0762] a) comprises an ECM degrading domain that comprises a hyaluronidase, and:

[0763] i) comprises or consists of an amino acid sequence of SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45; or of an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45; and / orii) is encoded by a nucleotide sequence of SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46; or

[0764] b) comprises an ECM degrading domain that comprises a neuraminidase, and:

[0765] i) comprises or consists of an amino acid sequence of SEQ ID NO: 5, SEQ ID NO: 106, SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 114or an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 5, SEQ ID NO: 106, SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 112, or SEQ ID NO: 114; and / or ii) is encoded by a nucleotide sequence of SEQ ID NO: 6, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 6, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, or SEQ ID NO: 115.

[0766] 12a. The recombinant bacterial cell of any of paragraphs 1-12, wherein the heterologous extracellular matrix (ECM) degrading polypeptide:

[0767] a) is secreted from the cell;

[0768] b) is displayed on the surface of the cell; or

[0769] c) is expressed in the cell.

[0770] 12b. The recombinant bacterial cell of paragraphs l-12a, wherein the heterologous extracellular matrix (ECM) degrading polypeptide is secreted from the cell.

[0771] 12c. The recombinant bacterial cell of paragraphs ll-12b, wherein the secretion domain is not:

[0772] a) PhoA;

[0773] b) OmpF;

[0774] c) CvaC;d) TorA;

[0775] e) FdnG;

[0776] f) DmsA; and / or

[0777] h) HlyA.

[0778] 13. The recombinant bacterial cell of any of paragraphs l-12b, wherein the bacterial cell comprises a nucleic acid comprising a nucleotide sequence encoding a transcriptional regulatory polypeptide; optionally

[0779] wherein the transcriptional regulatory polypeptide and the heterologous extracellular matrix (ECM) degrading polypeptide are comprised by the same nucleic acid.

[0780] 14. The recombinant bacterial cell of paragraph 13, wherein where the secretion domain is a FliC signal sequence, the transcriptional regulatory peptide is selected from the group comprising or consisting of: a FliC repressor; optionally wherein the FliC repressor is a GadE repressor; optionally

[0781] wherein the transcriptional regulatory peptide:

[0782] a) comprises or consists of an amino acid sequence of SEQ ID NO: 49; or of an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 49; and / or

[0783] b) is encoded by a nucleotide sequence of SEQ ID NO: 50, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 50.

[0784] 15. The recombinant bacterial cell of any one of paragraphs 13-14, wherein the transcriptional regulatory peptide is selected from the group comprising or consisting of:

[0785] a) FNR;

[0786] b) NagC;

[0787] c) CRP;

[0788] d) EutR; and / or

[0789] e) LldR.

[0790] 15a. The recombinant bacterial cell of any of paragraphs 1-15, wherein the hypoxiainducible promoter comprises a modified regulatory element selected from the group comprising or consisting of: a modified RNA polymerase binding sequence; a modified operator sequence; or any combination thereof.15b. The recombinant bacterial cell of any of paragraphs l-15a, wherein the pH-inducible promoter comprises a modified regulatory element selected from the group comprising or consisting of: a modified RNA polymerase binding sequence; a modified operator sequence; or any combination thereof.

[0791] 16. The recombinant bacterial cell of any one of paragraphs l-15b, wherein the cell is an auxotrophic cell; optionally

[0792] wherein the cell is not capable of endogenously expressing an essential gene; optionally

[0793] wherein the essential gene has been deleted from the genome of the cell.

[0794] 17. The recombinant bacterial cell of paragraph 16, wherein the nucleic acid further comprises a complementation cassette comprising a nucleotide sequence encoding the essential gene operably linked to a complementation promoter.

[0795] 18. The recombinant bacterial cell of paragraph 16 or 17, wherein the essential gene is selected from the group comprising or consisting of: dapD, infA, or a homologue thereof; optionally

[0796] wherein the essential gene is dapD or a homologue thereof; optionally wherein the essential gene comprises or consists of a nucleotide sequence of: SEQ ID NO: 268 or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 268.

[0797] 19. The recombinant bacterial cell of any one of paragraphs 17-18, wherein the complementation promoter is a constitutive promoter; optionally

[0798] wherein the complementation promoter is selected from the group comprising or consisting of: PdapD, lacZ, uvrA, and aroL; optionally

[0799] wherein the complementation promoter is PdapD; optionally

[0800] wherein the complementation promoter comprises or consisted of a nucleotide sequence of: SEQ ID NO 269; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 269.

[0801] 20. The recombinant bacterial cell of any one of paragraphs 1-19, wherein:

[0802] a) the nucleic acid not comprise a nucleotide sequence encoding an antibiotic resistance marker; and / or

[0803] b) the recombinant bacterial cell is not resistant to an antibiotic.21. The recombinant bacterial cell of any of paragraphs 1-20, wherein:

[0804] i) the cell is a non-pathogenic bacterial cell, a commensal cell, or a probiotic cell;

[0805] ii) the cell is a facultatively anaerobic cell;

[0806] and / or

[0807] iii) the cell is:

[0808] a) a gram-negative bacterial cell; optionally

[0809] wherein the gram-negative bacterial cell is selected from the group comprising or consisting of: an Escherichia coli cell;

[0810] optionally

[0811] wherein the gram-negative bacterial cell is an Escherichia coli K12 cell;

[0812] optionally wherein the Escherichia coli K12 cell is Escherichia coli K12 MG1655 cell;

[0813] or

[0814] b) a gram-positive bacterial cell; optionally

[0815] wherein the gram-positive bacterial cell is a lactic acid bacterial cell; optionally wherein the lactic acid bacterial cell is selected from the group comprising or consisting of: a Lactococcus lactis cell, a Lactobacillus cell, and a Bifidobacterium cell

[0816] optionally

[0817] wherein the gram-positive bacterial cell is a Lactococcus lactis MG1363 cell.

[0818] 21a. The recombinant bacterial cell of any of paragraphs 1-21, wherein the cell is not an attenuated bacterial cell.

[0819] 21b. The recombinant bacterial cell of any of paragraphs l-21a, wherein the cell is not:

[0820] a) a Salmonella cell; optionally a Salmonella bongori cell, a Salmonella cholreaesuis cell, a Salmonella enterica cell, a Salmonella enteritidis cell, a Salmonella paratyphi cell, a Salmonella typhi cell, or a Salmonella typhimurium cell; b) a Vibrio cell; optionally a Vibrio cholerae cell or a Vibrio fischeri cell; c) a Shigella cell; optionally a Shigella boydii cell, a Shigella dysenteriae cell, a Shigella flexneri cell, or a Shigella sonnei cell;

[0821] d) a Lactobacillus cell; optionally a Lactobacillus bulgaricus cell;

[0822] e) a Listeria cell; optionally a Listeria monocytogenes cell;

[0823] f) an Enterococcus cell; optionally an Enterococcus faecium cell;

[0824] g) a Streptococcus cell; optionally a Streptococcus pyogenes cell;h) a pathogenic Escherichia coli cell; optionally an Enteroinvasive Escherichia coli (EIEC) cell, an Adherent-Invasive Escherichia coli (AIEC) cell, an Enteroaggregative Escherichia coli (EAEC) cell, a Shiga Toxin-producing Escherichia coli (STEC) cell, an Enterotoxigenic Escherichia coli (ETEC) cell, an Enterohemorrhagic Escherichia coli (EHEC) cell, or an Extra intestinal Pathogenic Escherichia coli (Ex PEC) cell;

[0825] i) an Escherichia coli Nissile cell.

[0826] 21c. The recombinant bacterial cell of any one of paragraphs l-21b, wherein:

[0827] a) the cell is not a genotoxic cell;

[0828] b) the cell is a non-genotoxic cell; and / or

[0829] c) the cell does not produce a genotoxin.

[0830] 21d. The recombinant bacterial cell of any one of paragraphs l-21c, wherein the cell does not produce a cytotoxin.

[0831] 21e. The recombinant bacterial cell of any one of paragraphs l-21d, wherein:

[0832] a) the cell does not produce O-antigen;

[0833] b) the cell does not comprise O-antigen; and / or

[0834] c) the cell does not comprise a bacterial capsule.

[0835] 21f. The recombinant bacterial cell of any one of paragraphs l-21e, wherein:

[0836] a) the cell does not comprise an active prophage;

[0837] b) the cell comprises an inactive prophage; and / or

[0838] c) the cell comprises a non-functional prophage.

[0839] 21g. The recombinant bacterial cell of any of paragraphs l-21f, wherein the heterologous extracellular matrix (ECM) degrading polypeptide does not localise to an intracellular compartment of the cell.

[0840] 21h. The recombinant bacterial cell of any of paragraphs l-21g, wherein the heterologous extracellular matrix (ECM) degrading polypeptide does not localise to a membrane or cell wall of the cell.

[0841] 21 i. The recombinant bacterial cell of any of paragraphs 2-21h, wherein the nucleic acid further comprises a nucleotide sequence encoding a transcriptional regulatory element.21j. The recombinant bacterial cell of paragraph 21i, wherein the nucleotide sequence encoding the heterologous extracellular matrix degrading (ECM) polypeptide is operably linked to the transcriptional regulatory element.

[0842] 21k. The recombinant bacterial cell of paragraph 21i or 21j, wherein the transcriptional regulatory element is selected from the group comprising or consisting of: a terminator sequence, a repressor binding sequence, an insulator, an operator sequence, or any combination thereof.

[0843] 211. The recombinant bacterial cell of paragraph 21k, wherein:

[0844] a) the terminator is selected from the group comprising or consisting of: rrnB T1 / T2; optionally

[0845] wherein the terminator sequence has a sequence of SEQ ID NO: 51; b) the repressor binding sequence is selected from the group comprising or consisting of: FNR; optionally

[0846] wherein the repressor binding sequence has a sequence of SEQ ID NO: 52; and / or

[0847] c) the insulator sequence is selected from the group comprising or consisting of: RiboJ; optionally

[0848] wherein the insulator sequence has a sequence of SEQ ID NO: 53.

[0849] 22. The recombinant bacterial cell of any of paragraphs 1-211, wherein:

[0850] a) the nucleic acid is selected from the group comprising or consisting of: a plasmid, a phagemid, and a bacterial artificial chromosome (BAC);

[0851] b) the nucleic acid is a plasmid;

[0852] or

[0853] c) the nucleic acid integrated into the genome of the cell.

[0854] 23. The recombinant bacterial cell of any of paragraphs 1-22, wherein the cell is capable of accumulating in a target tissue or tumour.

[0855] 24. A tumour-inducible promoter, wherein the tumour-inducible promoter is a promoter according to any of paragraphs 1, 2, and 4-15.

[0856] 25. A nucleic acid comprising a nucleotide sequence of the tumour-inducible promoter of paragraph 24.26. A nucleic acid comprising a nucleotide sequence encoding an extracellular matrix (ECM) degrading polypeptide operably linked to a tumour-inducible promoter.

[0857] 27. The nucleic acid of paragraph 26:

[0858] i) wherein the extracellular matrix (ECM) degrading polypeptide is the heterologous extracellular matrix (ECM) degrading polypeptide according to any of paragraphs 1-3 and 10-12;

[0859] ii) wherein the tumour-promoter is the tumour-inducible promoter of any one of paragraphs 1, 2, and 4-15 and 25;

[0860] iii) the nucleic acid is selected from the group comprising or consisting of: a plasmid, a phagemid, and a bacterial artificial chromosome (BAC);

[0861] iv) wherein the nucleic acid is a plasmid.

[0862] 28. A cell comprising the tumour-inducible promoter of paragraph 28, and / or the nucleic acid of any one of paragraphs 25-27.

[0863] 29. The cell of paragraph 28, wherein the cell is a recombinant bacterial cell; optionally wherein the cell is a recombinant bacterial cell according to any of paragraphs 1-23.

[0864] 29a. The cell of paragraph 29, wherein the nucleic acid is:

[0865] a) extrachromosomal; or

[0866] b) integrated into the genome of the cell.

[0867] 30. The recombinant bacterial cell of any of paragraphs 1-23 for use in medicine.

[0868] 31. The recombinant bacterial cell of any of paragraphs l-23for use as a medicament.

[0869] 32. The recombinant bacterial cell of any of paragraphs 1-23 for use in treating cancer.

[0870] 33. The cell of any of paragraphs 28-29a62 for use in medicine.

[0871] 34. The cell of any of paragraphs 28-29a for use as a medicament.

[0872] 35. The cell of any of paragraphs 28-29a for use in treating cancer.

[0873] 36. The recombinant bacterial cell for use of any of paragraphs 30-32 or cell for use of any of paragraphs 33-35, wherein the use comprises administering an effective dose of the cell to a subject that has cancer.37. The recombinant bacterial cell for use according to paragraph 32 or 36, or the cell for use of any of paragraph 35 or 36, wherein the cancer is a tumour solid cancer, optionally wherein the cancer is an immune-excluded solid tumour cancer and / or an immune-excluded tumour.

[0874] 38. The recombinant bacterial cell for use according to paragraph 32, 36, or 37, or cell for use according to any of paragraphs 68-70, wherein:

[0875] a) the cancer is a cancer that comprises extracellular matrix; optionally wherein the cancer is selected from the group comprising or consisting of: triple-negative breast cancer, microsatellite stable colon cancer, and pancreatic cancer; and / or b) the cancer is not a cancer that does not comprise extracellular matrix; optionally wherein the cancer is not a blood cancer; optionally wherein the cancer is not a cancer selected from the group comprising or consisting of: leukaemia, myeloma, and lymphoma.

[0876] 39. A composition comprising the recombinant bacterial cell of paragraphs 1-23 or the cell of paragraphs 28-29a, optionally wherein the composition is a pharmaceutical composition.

[0877] 40. The composition of paragraph 39 for use in medicine.

[0878] 41. The composition of paragraph 39 for use as a medicament.

[0879] 42. The composition of paragraph 39 for use in a method of treating cancer.

[0880] 43. The composition for use of any of paragraphs 40-42, wherein the use comprises administering an effective dose of the composition to a subject.

[0881] 44. The composition for use of any of paragraphs 40-43, wherein the cancer is a solid tumour cancer, optionally wherein the cancer is an immune-excluded solid cancer.

[0882] 45. The composition for use of any of paragraphs 40-43, wherein the cancer is a cancer selected from the group comprising or consisting of: triple-negative breast cancer, microsatellite stable colon cancer, and pancreatic cancer.

[0883] 46. A method for degrading extracellular matrix (ECM) in a tissue, comprising contacting the tissue with the recombinant bacterial cell of any of paragraphs 1-23, the cell of paragraphs 28-29a, or the composition of paragraph 39.47. The method of paragraph 46, wherein the method is an in vivo method.

[0884] 48. The method of paragraph 46 or 47, wherein the method comprises administering the recombinant bacterial cell, the cell, or the composition to a subject.

[0885] 49. The method of paragraph 48, wherein the subject has a disease.

[0886] 50. The method of paragraph 49, wherein the disease is cancer; optionally wherein the cancer is a solid tumour cancer, optionally wherein the cancer is an immune-excluded solid cancer.

[0887] 51. The method of paragraph 50, wherein:

[0888] a) the cancer is a cancer that comprises extracellular matrix; optionally wherein the cancer is selected from the group comprising or consisting of: triple-negative breast cancer, microsatellite stable colon cancer, and pancreatic cancer; and / or b) the cancer is not a cancer that does not comprise extracellular matrix; optionally wherein the cancer is not a blood cancer; optionally wherein the cancer is not a cancer selected from the group comprising or consisting of: leukaemia, myeloma, and lymphoma.

[0889] 52. The method of paragraph 46, wherein the method is an in vitro method.

[0890] 53. The recombinant bacterial cell for use of any of paragraphs 30-32, 66-68, cell for use of any of paragraphs 33-68, composition for use of any of paragraphs 40-45, or the method of any of paragraphs 46-52, wherein the subject is a human subject.

[0891] 54. Use of the recombinant bacterial cell of any of paragraphs 1-23 the cell of paragraphs 28-29a, or the composition of paragraph 39 to degrade extracellular matrix (ECM).

[0892] 55. Use of paragraph 54, wherein the use is an in vitro use or an in vivo use.

[0893] 56. Use of paragraphs 54-55, wherein the use comprises administering the recombinant bacterial cell, the cell, or the composition to a subject.

[0894] 57. The recombinant bacterial cell for use of any of paragraphs 30-32, 3638 and 53, cell for use of any of paragraphs 33-38 or 53, composition for use of any paragraphs 4045 and 53, method of any of paragraphs 46-53, or use of any of paragraphs 44-46, whereinthe recombinant bacterial cell, the cell, or the composition is administered to a subject by a route selected from the group comprising or consisting of: oral administration, intraocular administration, intravenous administration, intraarterial administration, intraperitoneal administration, intramuscular administration, intratumoural administration, buccal administration, nasal administration, pulmonary administration.

[0895] 58. The recombinant bacterial cell for use, composition for use, method, or use of paragraph 57, wherein following administration the recombinant bacterial cell or cell accumulates in a target tissue or tumour in the subject.

[0896] 59. The recombinant bacterial cell for use, composition for use, method, or use of paragraph 58, wherein the recombinant bacterial cell or cell degrades the extracellular matrix (ECM) of the target tissue or tumour.

[0897] 60. The recombinant bacterial cell for use of any of paragraphs 30-32, 36-38, 53 and 57-59, cell for use of any of paragraphs 33-38, 53, and 57-59, composition for use of any of paragraphs 40-45, 53, and 57-59, method of any of paragraphs 46-53, 57-59, or use of any of paragraphs 54-59, 43-44, wherein the recombinant bacterial cell, cell, or composition is:

[0898] a) administered prior to or subsequent to administration of a cancer therapeutic; and / or

[0899] b) co-administered with a cancer therapeutic.

[0900] 61. The recombinant bacterial cell for use, composition for use, method, or use of paragraph 60, wherein the cancer therapeutic is selected from the group comprising or consisting of:

[0901] a) an antibody (optionally selected from the group comprising or consisting of: an immune checkpoint inhibitor (ICI), T-cell engager, and a bispecific antibody or an antibody-drug conjugate), a cell therapy (optionally selected from the group comprising or consisting of: a T-cell, a CAR-T cell, and a T4 cell), a chemotherapy agent (optionally selected from the group comprising or consisting of: FOLFOX, Gemcitabine, Paclitaxel, Cisplatin, Epirubicin and Irinotecan) an Immunomodulator (optionally selected from the group comprising or consisting of: a cytokine and a chemokine), a vaccine (optionally an mRNA vaccine), a virus (optionally an oncolytic virus), and a second live biotherapeutic (optionally selected from the group comprising or consisting of: an engineered live biotherapeutic and a recombinant bacterial cell); and / orb) an antibody (optionally an immune checkpoint inhibitor antibody, bispecific antibody or an antibody-drug conjugate), a hormonal therapy, a targeted therapy, an immunotherapy, a cell therapy (optionally a tumour infiltrating lymphocyte (TIL), a lymphocyte with an engineered T cell receptor (TCR), a CAR T cell, or a Natural Killer cell), an immunomodulator (optionally a chemokine or a cytokine), a vaccine (optionally an mRNA vaccine), a virus (optionally an oncolytic virus), another luve biotherapeutic (optionally an engineered live biotherapeutic), a chemotherapy (optionally an alkylating agent (optionally temozolomide and carboplatin), an antimetabolite (optionally 5-FU or gemcitabine), an anti-tumour antibiotic (optionally doxorubicin), a topoisomerase inhibitor (optionally irinotecan), a mitotic inhibitor (optionally a taxane, like paclitaxel or docetaxel), a corticosteroid (optionally dexamethasone).

[0902] 62. A pharmaceutical composition comprising the recombinant bacterial cell of any of paragraphs 1-23 or the cell of any of paragraphs 28-29a, and a cancer therapeutic, for use in treating cancer.

[0903] 63. A combination comprising the recombinant bacterial cell of any of paragraphs 1-23 or cell of paragraphs 28-29a and a cancer therapeutic for use in treating cancer.

[0904] 64. A combination comprising a cancer therapeutic and the recombinant bacterial cell of any of paragraphs 1-23 or cell of paragraphs 28-29a for use in treating cancer.

[0905] 65. The pharmaceutical composition for use of paragraph 62, or combination for use of paragraph 63 or 64, wherein:

[0906] a) the cancer is a cancer that comprises extracellular matrix; optionally wherein the cancer is selected from the group comprising or consisting of: triple-negative breast cancer, microsatellite stable colon cancer, and pancreatic cancer; and / or b) the cancer is not a cancer that does not comprise extracellular matrix; optionally wherein the cancer is not a blood cancer; optionally wherein the cancer is not a cancer selected from the group comprising or consisting of: leukaemia, myeloma, and lymphoma.

[0907] 66. The pharmaceutical composition for use of paragraphs 62 or 65, or combination for use of any of paragraphs 63-65, wherein the cancer therapeutic is a cancer immunotherapy agent, optionally wherein the cancer immunotherapy agent is selected from the group comprising or consisting of:a) an antibody (optionally selected from the group comprising or consisting of: an immune checkpoint inhibitor (ICI), T-cell engager, and a bispecific antibody or an antibody-drug conjugate), a cell therapy (optionally selected from the group comprising or consisting of: a T-cell, a CAR-T cell, and a T4 cell), a chemotherapy agent (optionally selected from the group comprising or consisting of: FOLFOX, Gemcitabine, Paclitaxel, Cisplatin, Epirubicin and Irinotecan) an Immunomodulator (optionally selected from the group comprising or consisting of: a cytokine and a chemokine), a vaccine (optionally an mRNA vaccine), a virus (optionally an oncolytic virus), and a second live biotherapeutic (optionally selected from the group comprising or consisting of: an engineered live biotherapeutic and a recombinant bacterial cell); and / or

[0908] b) an antibody (optionally an immune checkpoint inhibitor antibody, bispecific antibody or an antibody-drug conjugate), a hormonal therapy, a targeted therapy, an immunotherapy, a cell therapy (optionally a tumour infiltrating lymphocyte (TIL), a lymphocyte with an engineered T cell receptor (TCR), a CAR T cell, or a Natural Killer cell), an immunomodulator (optionally a chemokine or a cytokine), a vaccine (optionally an mRNA vaccine), a virus (optionally an oncolytic virus), another live biotherapeutic (optionally an engineered live biotherapeutic), a chemotherapy (optionally an alkylating agent (optionally temozolomide and carboplatin), an antimetabolite (optionally 5-FU or gemcitabine), an anti-tumour antibiotic (optionally doxorubicin), a topoisomerase inhibitor (optionally irinotecan), a mitotic inhibitor (optionally a taxane, like paclitaxel or docetaxel), a corticosteroid (optionally dexamethasone).

[0909] 67. The composition for use of any of paragraphs 62, 65, or 66, or combination for use of any of paragraphs 63-66, wherein the recombinant bacterial cell or cell is administered to a subject; optionally wherein:

[0910] a) the recombinant bacterial cell or cell is administered prior to or subsequent to administration of the cancer therapeutic; and / or

[0911] b) the recombinant bacterial cell or cell is co-administered with the cancer therapeutic.

[0912] 68. The composition for use of any of paragraphs 62 or 65-67, or combination for use of any of paragraphs 63-67, wherein the composition or combination for use is administered to a subject by a route selected from the group comprising or consisting of: oral administration, intraocular administration, intravenous administration, intraarterial administration, intraperitoneal administration, intramuscular administration, intratumoural administration, buccal administration, nasal administration, pulmonary administration.69. The composition for use of any of paragraphs 62 or 65-68, or combination for use of any of paragraphs 63-68, wherein following administration the recombinant bacterial cell or cell accumulates in a target tissue or tumour in the subject.

[0913] 70. The composition for use or combination for use of paragraph 69, wherein the recombinant bacterial cell or cell degrades the extracellular matrix (ECM) of the target tissue or tumour.

[0914] 71. The recombinant bacterial cell for use, composition for use, method, or use of paragraph 60, or the composition for use or combination for use of paragraph 70, wherein the degradation of the extracellular matrix (ECM) of the target tissue or tumour allows the cancer therapeutic to contact and / or enter the tumour;

[0915] optionally wherein said contacting and / or entering the tumour is required for the therapeutic effect of the cancer therapeutic.

[0916] 72. A kit of parts, comprising:

[0917] a) the recombinant bacterial cell of any of paragraphs 1-23;

[0918] b) the tumour-24; c) the nucleic acid of any of paragraphs 25-27; e) the cell of any of paragraphs 28-29a;

[0919] f) the composition of any of paragraphs 39-45;

[0920] g) a therapeutic agent, optionally an anti-cancer therapeutic.

[0921] Figure legends

[0922] Figure 1 - A) Luminescence measurement of luminescent E. coli K12 MG1655 using Photoimager Optima In Vivo Imaging System, showing successful generation of a luminescent strain. B) Bodyweight of mice administered luminescent E. coli K12 MG1655 intravenously at doses of 106, 107, and 108CFU, heat-killed bacteria, or vehicle only.

[0923] Figure 2 - Bioluminescence imaging using Photoimager Optima In Vivo Imaging System, showing localisation of luminescent E. coli K12 MG1655 to tumours in the mice.

[0924] Figure 3 - A) Bioluminescence imaging using Photoimager Optima In Vivo Imaging System, showing localisation of luminescent E. coli K12 MG1655 to tumours in the...

Claims

Claims1. A recombinant bacterial cell capable of expressing a heterologous extracellular matrix (ECM) degrading polypeptide that degrades an extracellular matrix component, wherein:the cell comprises a nucleic acid comprising a nucleotide sequence that encodes the heterologous extracellular matrix (ECM) degrading polypeptide and a promoter;wherein the nucleotide sequence that encodes the heterologous extracellular matrix (ECM) degrading polypeptide is operably linked to the promoter; andthe promoter is a tumour-inducible promoter.

2. The recombinant bacterial cell of claim 1, wherein the heterologous extracellular matrix (ECM) degrading polypeptide is secreted from the cell.

3. A recombinant bacterial cell capable of expressing a heterologous extracellular matrix (ECM) degrading polypeptide that degrades an extracellular matrix component, wherein the cell comprises a nucleic acid comprising a nucleotide sequence that encodes the heterologous extracellular matrix (ECM) degrading polypeptide,wherein the heterologous extracellular (ECM) degrading polypeptide is secreted from the cell.

4. The recombinant bacterial cell of claim 3, wherein the nucleic acid further comprises a promoter; optionallywherein the nucleotide sequence encoding the heterologous extracellular matrix degrading (ECM) polypeptide is operably linked to the promoter and wherein the promoter is a tumour-inducible promoter.

5. The recombinant bacterial cell of any one of claims 1, 2, and 4 wherein the tumourinducible promoter is:a) a hypoxia-inducible promoter; optionally wherein the hypoxia-inducible promoter comprises:i) an FNR binding motif; and / orii) an fnrS promoter or subsequence thereof; and / oriii) a grcA promoter or subsequence thereof;b) an N-acetylglucosamine-inducible promoter; optionally wherein the N- acetylglucosamine-inducible promoter comprises a NagC binding motif;c) an ethanolamine-inducible promoter; optionally wherein the ethanolamine- inducible promoter comprises a EutR binding motif;d) a lactate-inducible promoter; optionally wherein the lactate-inducible promoter comprises an IldR binding motif;e) a pH-inducible promoter;f) a cAMP-inducible promoter; optionally wherein the cAMP-inducible promoter comprises a CRP binding motif; optionallyg) a hypoxia- and N-acetylglucosamine-inducible promoter; optionally wherein the hypoxia- and N-acetylglucosamine-inducible promoter comprises a NagC binding motif and:i) an FNR binding motif; and / orii) an fnrS promoter or subsequence thereof; and / oriii) a grcA promoter or subsequence thereof;h) a hypoxia- and ethanolamine-inducible promoter; optionally wherein the hypoxia- and ethanolamine-inducible promoter comprises a EutR binding motif and:i) an FNR binding motif; and / orii) an fnrS promoter or subsequence thereof; and / oriii) a grcA promoter or subsequence thereof;i) a hypoxia- and lactate-inducible promoter; optionally wherein the hypoxia-and lactate-inducible promoter comprises an IldR binding motif and:i) an FNR binding motif; and / orii) an fnrS promoter or subsequence thereof; and / oriii) a grcA promoter or subsequence thereof;j) a hypoxia- and pH-inducible promoter; optionallywherein the hypoxia- and pH-inducible promoter comprises:i) an FNR binding motif; and / orii) an fnrS promoter or subsequence thereof; and / oriii) a grcA promoter or subsequence thereof;k) a hypoxia- and cAMP-inducible promoter; optionally wherein the hypoxia-and cAMP-inducible promoter comprises a CRP binding motif and:i) an FNR binding motif; and / orii) an fnrS promoter or subsequence thereof; and / oriii) a grcA promoter or subsequence thereof;l) a pH- and lactate-inducible promoter; optionally wherein the pH- and lactate-inducible promoter comprises an IldR binding motif;m) a hypoxia-, N-acetylglucosamine, and lactate-inducible promoter; optionally wherein the hypoxia-, N-acetylglucosamine, and lactate-inducible promoter comprises a NagC binding motif, an IldR binding motif, and:i) an FNR binding motif; and / orii) an fnrS promoter or subsequence thereof; and / oriii) a grcA promoter or subsequence thereof;n) a hypoxia-, cAMP, and lactate-inducible promoter; optionally wherein the hypoxia-, cAMP, and lactate-inducible promoter comprises a CRP binding motif, an IldR binding motif, and:i) an FNR binding motif; and / orii) an fnrS promoter or subsequence thereof; and / oriii) a grcA promoter or subsequence thereof;o) an ethanolamine- and lactate-inducible promoter; optionally wherein the ethanolamine- and lactate-inducible promoter comprises a EutR binding motif and an IldR binding motif;p) an ethanolamine-, lactate-, and N-acetylglucosamine-inducible promoter; optionally wherein the ethanolamine-, lactate-, and N-acetylglucosamine-inducible promoter comprises a EutR binding motif, an IldR binding motif, and a NagC binding motif;q) an ethanolamine-, lactate-, and hypoxia-inducible promoter; optionally wherein the ethanolamine-, lactate-, and hypoxia-inducible promoter comprises a EutR binding motif, an IldR binding motif, and:i) an FNR binding motif; and / orii) an fnrS promoter or subsequence thereof; and / oriii) a grcA promoter or subsequence thereof;and / orr) an ethanolamine-, lactate-, hypoxia-, and N-acetylglucosamine-inducible promoter; optionally wherein the ethanolamine-, lactate-, hypoxia- and N- acetylglucosamine-inducible promoter comprises a EutR binding motif, an IldR binding motif, a NagC binding motif and:i) an FNR binding motif; and / orii) an fnrS promoter or subsequence thereof; and / oriii) a grcA promoter or subsequence thereof.

6. The tumour-inducible promoter of claim 5, wherein:a) the FNR binding motif comprises or consists of a nucleotide sequence of: SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, or SEQ ID NO: 244;b) the fnrS promoter or subsequence thereof comprises or consists of a nucleotide sequence of: SEQ ID NO: 250 or SEQ ID NO: 257, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 250 or SEQ ID NO: 257;c) the NagC binding motif comprises or consists of a nucleotide sequence of: SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 239, or SEQ ID NO: 259, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 239, or SEQ ID NO: 259;d) the EutR binding motif comprises or consists of a nucleotide sequence of: SEQ ID NO: 261 or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 261;e) the IldR binding motif comprises or consists of a nucleotide sequence of: SEQ ID NO: 248, SEQ ID NO: 249, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249;f) the grcA promoter or subsequence thereof comprises or consists of a nucleotide sequence of: SEQ ID NO: 253 or SEQ ID NO: 254, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 238 or SEQ ID NO: 249;and / org) CRP binding motif comprises or consists of a nucleotide sequence of: SEQ ID NO: 245 or SEQ ID NO: 246; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 245 or SEQ ID NO: 246.

7. The recombinant bacterial cell of any one of claims 1, 2, and 4-6, wherein the tumour-inducible promoter comprises:a) a discriminator; optionallywherein the discriminator comprises or consists of a nucleotide sequence of SEQ ID NO: 229, SEQ ID NO: 230, SEQ ID NO: 231, SEQ ID NO: 232, SEQ ID NO: 233, SEQ ID NO: 234, SEQ ID NO: 235, SEQ ID NO: 236, SEQ ID NO: 237, SEQ IDNO: 238, SEQ ID NO: 240, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 229, SEQ ID NO: 230, SEQ ID NO: 231, SEQ ID NO: 232, SEQ ID NO: 233, SEQ ID NO: 234, SEQ ID NO: 235, SEQ ID NO: 236, SEQ ID NO: 237, SEQ ID NO: 238, or SEQ ID NO: 240; optionallywherein the discriminator is a "high GC" discriminator; optionally wherein the "high GC" discriminator comprises or consists of a nucleotide sequence of SEQ ID NO: 232 or SEQ ID NO: 233;b) a -10 / -35 spacer; optionallywherein the 10 / -35 spacer is selected from the group comprising or consisting of: SEQ ID NO: 217, SEQ ID NO: 218, SEQ ID NO: 219, SEQ ID NO: 220, SEQ ID NO 221, SEQ ID NO: 222, SEQ ID NO: 223, SEQ ID NO: 224, SEQ ID NO: 225, or SEQ ID NO: 226, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 217, SEQ ID NO: 218, SEQ ID NO: 219, SEQ ID NO: 220, SEQ ID NO 221, SEQ ID NO: 222, SEQ ID NO: 223, SEQ ID NO: 224, SEQ ID NO: 225, or SEQ ID NO: 226;c) a TUSP binding site; optionallywherein the TUSP binding site comprises or consists of a nucleotide sequence of SEQ ID NO: 227, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 227;d) a TGIF sequence; optionallywherein the TGIF sequence comprises or consists of a nucleotide sequence of SEQ ID NO: 230, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 230; ore) a terminator sequence; optionallywherein the terminator sequence comprises or consists of a nucleotide sequence of SEQ ID NO: 247, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 247;f) a ribosome binding site (RBS); optionallywherein the ribosome binding site (RBS) comprises or consists of a nucleotide sequence of SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ IDNO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, or SEQ ID NO: 105; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, or SEQ ID NO: 105.

8. The recombinant bacterial cell of any one of claims 1, 2, and 4-7, wherein the tumour-inducible promoter comprises or consists of a nucleotide sequence of SEQ ID NO 160, SEQ ID NO: 275, SEQ ID NO: 182, SEQ ID NO: 276, SEQ ID NO: 190, SEQ ID NO: 277, SEQ ID NO: 128, SEQ ID NO: 278, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO; 125, SEQ ID NO: 126, SEQ ID NO:: 27,, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO; 135, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO; 145, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO; 155, SEQ ID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO; 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO; 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO: 179, SEQ ID NO: 180, SEQ ID NO: 181, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO; 185, SEQ ID NO: 186, SEQ ID NO: 187, SEQ ID NO: 188, SEQ ID NO: 189, SEQ ID NO: 191, SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO; 195, SEQ ID NO: 196, SEQ ID NO: 197, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO; 205, SEQ ID NO: 206, SEQ ID NO:207, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210, or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO 160, SEQ ID NO 275, SEQ ID NO 182, SEQ ID NO: 276, SEQ ID NO: 190, SEQ ID NO: 277, SEQ ID NO 128, SEQ ID NO 278, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO 124, SEQ ID NO 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 129, SEQ ID NO 130, SEQ ID NO 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO; 135, SEQ ID NO 136, SEQ ID NO 139, SEQ ID NO: 140, SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO; 145, SEQ ID NO 146, SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO; 155, SEQ ID NO 156, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, SEQ ID NO 161, SEQ ID NO 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO; 165, SEQ ID NO 166, SEQ ID NO 167, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO 171, SEQ ID NO 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO; 175, SEQ ID NO 176, SEQ ID NO 177, SEQ ID NO: 178, SEQ ID NO: 179, SEQ ID NO: 180, SEQ ID NO 181, SEQ ID NO 183, SEQ ID NO: 184, SEQ ID NO; 185, SEQ ID NO: 186, SEQ ID NO 187, SEQ ID NO 188, SEQ ID NO: 189, SEQ ID NO: 191, SEQ ID NO: 192, SEQ ID NO 193, SEQ ID NO 194, SEQ ID NO; 195, SEQ ID NO: 196, SEQ ID NO: 197, SEQ ID NO 198, SEQ ID NO 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO 203, SEQ ID NO 204, SEQ ID NO; 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210.

9. The recombinant bacterial cell of any one of claims 1, 2, and 4-8, wherein the tumour-inducible promoter is:a) a hypoxia-inducible promoter, optionally wherein the hypoxia-inducible promoter comprises or consists of a nucleotide sequence of:i) SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 128, SEQ ID NO: 278, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ D NO: 132, SEQ UD NO: 133, SEQ ID NO 135, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO; 155, SEQ ID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO; 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172,SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO; 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO: 179, SEQ ID NO: 180, SEQ ID NO: 189, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO; 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 128, SEQ ID NO: 278, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 54, SEQ ID NO: 55, or SEQ ID NO: 56, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ D NO: 132, SEQ ID NO: 133, SEQ ID NO 135, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: 150, SEQ ID NO: 179, SEQ ID NO: 180, SEQ ID NO: 189, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO; 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, or SEQ ID NO: 210; and / or ii) SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 128, SEQ ID NO: 278, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127,, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ D NO: 132, SEQ UD NO: 133, SEQ ID NO 135, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: SEQ ID NO: 150, SEQ ID NO: 179, or SEQ ID NO: 180; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 128, SEQ ID NO: 278, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ D NO: 132, SEQ UD NO: 133, SEQ ID NO 135, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: SEQ ID NO: 150, SEQ ID NO: 179, or SEQ ID NO: 180;b) an N-acetylglucosamine-inducible promoter; optionally wherein the N-acetylglucosamine-inducible promoter comprises or consists of a nucleotide sequence of:the N-acetylglucosamine-inducible promoter comprises a sequence of SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 169, SEQ ID NO: 172,SEQ ID NI: 173, SEQ ID NO: 194, SEQ ID NO: 196, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, or SEQ ID NO: 207; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 169, SEQ ID NO: 172, SEQ ID NI: 173, SEQ ID NO: 194, SEQ ID NO: 196, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, or SEQ ID NO: 207; c) an ethanolamine-inducible promoter; optionally wherein the ethanolamine-inducible promoter comprises or consists of a nucleotide sequence of:i) SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, or SEQ ID NO: 197; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, or SEQ ID NO: 197; and / orii) SEQ ID NO: 192 or SEQ ID NO: 197; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 192 or SEQ ID NO: 197;d) a lactate-inducible promoter; optionally wherein the lactate-inducible promoter comprises or consists of a nucleotide sequence of:SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 190, SEQ ID NO: 277, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO: 155, SEQ ID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO; 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO; 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO; 189, SEQ ID NO: 191, SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, or SEQ ID NO: 210; or a sequence is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 190, SEQ ID NO: 277, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO: 155, SEQ ID NO: 156, SEQ ID NO: 157, SEQ IDNO: 158, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO; 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO; 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO; 189, SEQ ID NO: 191, SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, or SEQ ID NO: 210;e) a pH-inducible promoter; optionally wherein the pH-inducible promoter comprises or consists of a nucleotide sequence of:i) SEQ ID NO: 182, SEQ ID NO: 276, SEQ ID NO: 190, SEQ ID NO: 277, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO 181, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 187, SEQ ID NO: 188, or SEQ ID NO: 191; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 182, SEQ ID NO: 276, SEQ ID NO: 190, SEQ ID NO: 277, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, or SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO 181, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 187, SEQ ID NO: 188, or SEQ ID NO: 191; and / orii) SEQ ID NO: 182, SEQ ID NO: 276, SEQ ID NO: 190, SEQ ID NO: 277, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO 181, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 187; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 182, SEQ ID NO: 276, SEQ ID NO: 190, SEQ ID NO: 277, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO 181, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO: 185, SEQ ID NO: 186, or SEQ ID NO: 187;f) a cAMP-inducible promoter; optionally wherein the cAMP-inducible promoter comprises or consists of a nucleotide sequence of:SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 170, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 189, SEQ ID NO: 208, SEQ ID NO: 209, or SEQ ID NO: 210; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, atleast 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to: SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 170, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 189, SEQ ID NO: 208, SEQ ID NO: 209, or SEQ ID NO: 210;g) a hypoxia- and N-acetylglucosamine-inducible promoter; optionally wherein the hypoxia- and N-acetylglucosamine-inducible promoter comprises or consists of a nucleotide sequence of::i) SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 169, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 196, SEQ ID NO: 198, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 169, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 196, SEQ ID NO: 198, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207; and / orii) SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 198, or SEQ ID NO: 200; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 145, SEQ ID NO: 198, or SEQ ID NO: 200;h) a hypoxia- and ethanolamine-inducible promoter; optionally wherein the hypoxia- and ethanolamine-inducible promoter comprises or consists of a nucleotide sequence of:SEQ ID NO: 195 or SEQ ID NO: 196; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 195 or SEQ ID NO: 196;i) a hypoxia- and lactate-inducible promoter; optionally wherein the hypoxia- and lactate-inducible promoter comprises or consists of a nucleotide sequence of:i) SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 156, SEQ ID NO: 157, SEQ IDNO: 158, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO: 189, SEQ ID NO: 195, SEQ IDNO: 196, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, or SEQ ID NO: 207; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 156, SEQ ID NO: 157, SEQ IDNO: 158, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 170, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO: 189, SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, or SEQ ID NO: 207; and / orii) SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 156, SEQ ID NO: 157, SEQ IDNO: 158, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 171, SEQ ID NO: 174, SEQ ID NO: 175, or SEQ ID NO: 178; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 160, SEQ ID NO: 275, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 156, SEQ ID NO: 157, SEQ IDNO: 158, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 171, SEQ ID NO: 174, SEQ ID NO: 175, or SEQ ID NO: 178;j) a hypoxia- and pH-inducible promoter; optionally wherein the pH-inducible and hypoxia-inducible promoter comprises or consists of a nucleotide sequence of:SEQ ID NO: 73 or SEQ ID NO: 188; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 73 or SEQ ID NO: SEQ ID NO: 188;k) a hypoxia- and cAMP-inducible promoter; optionally wherein the hypoxia-and cAMP-inducible promoter comprises or consists of a nucleotide sequence of:i) SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 170, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 189, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%,at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 170, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 189, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210; and / orii) SEQ ID NO: 148, SEQ ID NO: 149; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 148, or SEQ ID NO: 149;l) a pH- and lactate-inducible promoter; optionally wherein the pH- and lactate-inducible promoter comprises or consists of a nucleotide sequence of:SEQ ID NO: 190, or SEQ ID NO: 277, or SEQ ID NO: 191; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 190, or SEQ ID NO: 277, or SEQ ID NO: 191;m) a hypoxia-, N-acetylglucosamine, and lactate-inducible promoter; optionally wherein:the hypoxia-, N-acetylglucosamine, and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 169, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 205, SEQ ID NO: 206, or SEQ ID NO: 206; or sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 169, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 205, SEQ ID NO: 206, or SEQ ID NO: 206; n) a hypoxia-, cAMP, and lactate-inducible promoter; optionally wherein: the hypoxia-, cAMP, and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 170, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 189, SEQ ID NO: 208, SEQ ID NO: 209, or SEQ ID NO: 210; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 170, SEQ ID NO: 176, SEQ ID NO: 177, SEQ ID NO: 189, SEQ ID NO: 208, SEQ ID NO: 209, or SEQ ID NO: 210;o) an ethanolamine- and lactate-inducible promoter; optionally wherein:i) the ethanolamine- and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, or SEQ ID NO: 196; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%,at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 195, or SEQ ID NO: 196; and / orii) the ethanolamine- and lactate-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 193 or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 193;p) an ethanolamine-, lactate-, and N-acetylglucosamine-inducible promoter; optionally wherein:i) the ethanolamine-, lactate-, and N-acetylglucosamine-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 194 or SEQ ID NO: 196; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 194 or SEQ ID NO: 196; and / orii) the ethanolamine-, lactate-, and N-acetylglucosamine-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 194; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 194;q) the tumour-inducible promoter is an ethanolamine-, lactate-, and hypoxiainducible promoter; optionally wherein:i) the ethanolamine-, lactate-, and hypoxia-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 195 or SEQ ID NO: 196; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 195 or SEQ ID NO: 196; and / orii) the ethanolamine-, lactate-, and hypoxia-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 195; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 195;and / orr) an ethanolamine-, lactate-, N-acetylglucosamine, and hypoxia-inducible promoter; optionally whereinthe ethanolamine-, lactate-, N-acetylglucosamine, and hypoxia-inducible promoter comprises or consists of a nucleotide sequence of: SEQ ID NO: 196; or a sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 196.

10. The recombinant bacterial cell of any one of claims 1-9, wherein:i) the extracellular matrix degrading polypeptide is:a) a hyaluronidase;b) a microbial hyaluronidase or a mammalian hyaluronidase;c) a bacterial hyaluronidase;d) chondroitin ABC lyase;e) a microbial chondroitin ABC lyase or a mammalian chondroitin ABC lyase;e) a bacterial chondroitin ABC lyase;f) neuraminidase;g) a microbial neuraminidase or a mammalian neuraminidase;h) a bacterial neuraminidase;i) PNGase;j) a microbial PNGase or a mammalian PNGase;k) a bacterial PNGase;optionally wherein the extracellular matrix component is a proteoglycan; ii) the ECM component is hyaluronic acid (HA) and the ECM degrading polypeptide is:a) a hyaluronidase;b) a microbial hyaluronidase or a mammalian hyaluronidase; and / or c) a bacterial hyaluronidase;iii) the ECM component is Versican and the ECM degrading polypeptide is: a) chondroitin ABC lyase;b) a microbial chondroitin ABC lyase or a mammalian chondroitin ABC lyase; and / orc) a bacterial chondroitin ABC lyase;iv) the ECM component is Versican and the ECM degrading polypeptide is: a) neuraminidase;b) a microbial neuraminidase or a mammalian neuraminidase; and / or c) a bacterial neuraminidase;v) the ECM component is Versican and the ECM degrading polypeptide is: a) PNGase;b) a microbial PNGase or a mammalian PNGase; and / or c) a bacterial PNGase;vi) the heterologous extracellular matrix (ECM) degrading polypeptide:a) is a hyaluronidase and comprises or consists of an amino acid sequence of SEQ ID NO: 1; is a chondroitin ABC Lyase and comprises or consists of an amino acid sequence of SEQ ID NO: 3 or 47; is a neuraminidase and comprises or consists of an amino acid sequence of SEQ ID NO: 5; or is a PNGase and comprises or consists of an amino acid sequence of SEQ ID NO: 7; or comprises or consists of an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, or SEQ ID NO: 47; and / orb) is encoded by a nucleotide sequence of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, or SEQ ID NO: 48 or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, or SEQ ID NO: 48;vii) the heterologous extracellular matrix (ECM) degrading polypeptide is a bacterial hyaluronidase; and / orviii) the heterologous extracellular matrix (ECM) degrading polypeptide is a bacterial hyaluronidase that:a) comprises or consists of an amino acid sequence of SEQ ID NO: 1; or of an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 1; and / or b) is encoded by a nucleotide sequence of SEQ ID NO: 2, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 2.

11. The recombinant bacterial cell of any one of claims 1-10, wherein the heterologous extracellular matrix (ECM) degrading polypeptide is a fusion polypeptide comprising an ECM degrading domain and:a) a secretion domain;b) two secretion domains;c) a cleavage domain; and / ord) a cleavage domain that is positioned between the ECM degrading domain and the secretion domain.

12. The recombinant bacterial cell of claim 11, wherein:i) the secretion domain is selected from the group comprising or consisting of: a PelB secretion signal sequence, a YebF carrier protein, a CtxB signal sequence, an AIDA-I autotransporter, a FliC signal sequence, and a Usp45 signal sequence, or any combination thereof;ii) the secretion domain:a) comprises or consists of an amino acid sequence of SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35; or of an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35; and / or b) is encoded by a nucleotide sequence of SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36;iii) the cleavage domain comprises a matrix metalloprotease (MMP) cleavage site, optionally wherein the cleavage domain:a) comprises or consists of an amino acid sequence of SEQ ID NO: 116; or of an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 116; and / or b) is encoded by a nucleotide sequence of SEQ ID NO: 117, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 117; and / oriv) the fusion polypeptide:a) comprises an ECM degrading domain that comprises a hyaluronidase, and:i) comprises or consists of an amino acid sequence of SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO:45; or of an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45; and / orii) is encoded by a nucleotide sequence of SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46; orb) comprises an ECM degrading domain that comprises a neuraminidase, and:i) comprises or consists of an amino acid sequence of SEQ ID NO: 5, SEQ ID NO: 106, SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 114or an amino acid sequence at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 5, SEQ ID NO: 106, SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 112, or SEQ ID NO: 114; and / or ii) is encoded by a nucleotide sequence of SEQ ID NO: 6, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 6, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, or SEQ ID NO: 115.

13. The recombinant bacterial cell of any of claims 1-12, wherein the bacterial cell comprises a nucleic acid comprising a nucleotide sequence encoding a transcriptional regulatory polypeptide; optionallywherein the transcriptional regulatory polypeptide and the heterologous extracellular matrix (ECM) degrading polypeptide are comprised by the same nucleic acid; optionally wherein:the transcriptional regulatory peptide is selected from the group comprising or consisting of:a) where the secretion domain is a FliC signal sequence, the transcriptional regulatory peptide is selected from the group comprising or consisting of: a FliC repressor; optionally wherein the FliC repressor is a GadE repressor; and / orb) FNR;c) NagC;d) CRP;e) EutR; and / orf) LldR.

14. The recombinant bacterial cell of any one of claims 1-13, wherein the cell is an auxotrophic cell; optionallywherein the cell is not capable of endogenously expressing an essential gene; optionallywherein the essential gene has been deleted from the genome of the cell; optionally wherein the essential gene is selected from the group comprising or consisting of: dapD, infA, or a homologue thereof; optionallywherein the essential gene is dapD or a homologue thereof; optionally wherein the essential gene comprises or consists of a nucleotide sequence encoded by SEQ ID NO: 268; or a nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 268.

15. The recombinant bacterial cell of claim 14, wherein the nucleic acid further comprises a complementation cassette comprising a nucleotide sequence encoding the essential gene operably linked to a complementation promoter; optionallywherein the complementation promoter is a constitutive promoter; optionally wherein the complementation promoter is selected from the group comprising or consisting of: PdapD, lacZ, uvrA, and aroL; optionallywherein the complementation promoter is PdapD; optionallywherein the complementation promoter has a sequence of SEQ ID NO 269; or nucleotide sequence that is at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 269.

16. The recombinant bacterial cell of any one of claims 1-15, wherein:a) the nucleic acid not comprise a nucleotide sequence encoding an antibiotic resistance marker;b) the recombinant bacterial cell is not resistant to an antibiotic; and / orc) the cell is capable of accumulating in a target tissue or tumour.

17. The recombinant bacterial cell of any of claims 1-16, wherein:i) the cell is a non-pathogenic bacterial cell, a commensal cell, or a probiotic cell;ii) the cell is a facultatively anaerobic cell;and / oriii) the cell is:a) a gram-negative bacterial cell; optionallywherein the gram-negative bacterial cell is selected from the group comprising or consisting of: an Escherichia coli cell;optionallywherein the gram-negative bacterial cell is an Escherichia coli K12 cell;optionally wherein the Escherichia coli K12 cell is Escherichia coli K12 MG1655 cell;orb) a gram-positive bacterial cell; optionallywherein the gram-positive bacterial cell is a lactic acid bacterial cell; optionally wherein the lactic acid bacterial cell is selected from the group comprising or consisting of: a Lactococcus lactis cell, a Lactobacillus cell, and a Bifidobacterium celloptionallywherein the gram-positive bacterial cell is a Lactococcus lactis MG1363 cell.

18. A tumour-inducible promoter, wherein the tumour-inducible promoter is a promoter according to any of claims 1, 2, and 4-9.

19. A nucleic acid comprising a nucleotide sequence of the tumour-inducible promoter of claim 18.

20. A nucleic acid comprising a nucleotide sequence encoding an extracellular matrix (ECM) degrading polypeptide operably linked to a tumour-inducible promoter, wherein:i) the extracellular matrix (ECM) degrading polypeptide is the heterologous extracellular matrix (ECM) degrading polypeptide according to any of claims 1-3 and 10-12;ii) the tumour-promoter is the tumour-inducible promoter of any one of claims 1, 2, and 4-9 and 18;iii) the nucleic acid is selected from the group comprising or consisting of: a plasmid, a phagemid, and a bacterial artificial chromosome (BAC); and / or iv) the nucleic acid is a plasmid.

21. A cell comprising the tumour-inducible promoter of claim 18, and / or the nucleic acid of any one of claims 19-20.

22. The cell of claim 21, wherein the cell is a recombinant bacterial cell; optionally wherein the cell is a recombinant bacterial cell according to any of claims 1-17.

23. The recombinant bacterial cell of any of claims 1-17 or cell of claim 21 or 22 for use in medicine.

24. The recombinant bacterial cell of any of claims 1-17 or cell of claim 21 or 22 for use in treating cancer.

25. The recombinant bacterial cell or the cell for use of any of claims 23-24, wherein:i) the cancer is a tumour solid cancer, optionally wherein the cancer is an immune-excluded solid tumour cancer and / or an immune-excluded tumour; ii) the cancer is a cancer that comprises extracellular matrix;iii) the cancer is a cancer that comprises extracellular matrix and is selected from the group comprising or consisting of: triple-negative breast cancer, microsatellite stable colon cancer, and pancreatic cancer; and / or iv) the cancer is not a cancer that does not comprise extracellular matrix; v) the cancer is not a cancer that does not comprise extracellular matrix and is not a blood cancer;vi) the cancer is not a cancer that does not comprise extracellular matrix, is not a blood cancer, and is not a cancer selected from the group comprising or consisting of: leukaemia, myeloma, and lymphoma; and / orvii) the recombinant bacterial cell or cell is:a) administered prior to or subsequent to administration of a cancer therapeutic;b) co-administered with a cancer therapeutic;c) co-administered with a cancer therapeutic, wherein the cancer therapeutic is selected from the group comprising or consisting of: an antibody (optionally selected from the group comprising or consisting of: animmune checkpoint inhibitor (ICI), T-cell engager, and a bispecific antibody or an antibody-drug conjugate), a cell therapy (optionally selected from the group comprising or consisting of: a T-cell, a CAR-T cell, and a T4 cell), a chemotherapy agent (optionally selected from the group comprising or consisting of: FOLFOX, Gemcitabine, Paclitaxel, Cisplatin, Epirubicin and Irinotecan) an Immunomodulator (optionally selected from the group comprising or consisting of: a cytokine and a chemokine), a vaccine (optionally an mRNA vaccine), a virus (optionally an oncolytic virus), and a second live biotherapeutic (optionally selected from the group comprising or consisting of: an engineered live biotherapeutic and a recombinant bacterial cell); ord) co-administered with a cancer therapeutic, wherein the cancer therapeutic is selected from the group comprising or consisting of: an antibody (optionally an immune checkpoint inhibitor antibody, bispecific antibody or an antibody-drug conjugate), a hormonal therapy, a targeted therapy, an immunotherapy, a cell therapy (optionally a tumour infiltrating lymphocyte (TIL), a lymphocyte with an engineered T cell receptor (TCR), a CAR T cell, or a Natural Killer cell), an immunomodulator (optionally a chemokine or a cytokine), a vaccine (optionally an mRNA vaccine), a virus (optionally an oncolytic virus), another live biotherapeutic (optionally an engineered live biotherapeutic), a chemotherapy (optionally an alkylating agent (optionally temozolomide and carboplatin), an anti-metabolite (optionally 5-FU or gemcitabine), an anti-tumour antibiotic (optionally doxorubicin), a topoisomerase inhibitor (optionally irinotecan), a mitotic inhibitor (optionally a taxane, optionally paclitaxel or docetaxel), a corticosteroid (optionally dexamethasone).

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