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WO 2016/124239 Al 11 August 2016 (11.08.2016) P O P C T (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2016/124239 Al 11 August 2016 (11.08.2016) P O P C T (51) International Patent Classification: AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, A61K 38/17 (2006.01) A61K 35/74 (2015.01) BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, A61K 39/02 (2006.01) A61P 17/00 (2006.01) DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, A61K 9/16 (2006.01) HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KN, KP, KR, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, (21) International Application Number: MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PCT/EP20 15/052345 PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, (22) International Filing Date: SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, 4 February 2015 (04.02.2015) TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (25) Filing Language: English (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, (26) Publication Language: English GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, (71) Applicant: AUREALIS OY [FI/FI]; Microkatu 1, FI- 70210 Kuopio (FI). TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, (72) Inventors: WIRTH, Thomas; Ritosentie 20 C 15, FI- LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, 70340 Kuopio (FI). YRJANHEIKKL Juha; Renkitie 20, SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, FI-70820 Kuopio (FI). SAMARANAYAKE, Haritha; GW, KM, ML, MR, NE, SN, TD, TG). Metsurintie 11 B 34, FI-70150 Kuopio (FI). Published: (74) Agent: WALCHER, Armin; Patentanwalte Louis Pohlau — with international search report (Art. 21(3)) Lohrentz, Postfach 30 55, 90014 Nurnberg (DE). — with sequence listing part of description (Rule 5.2(a)) (81) Designated States (unless otherwise indicated, for every kind of national protection available): AE, AG, AL, AM, o (54) Title: RECOMBINANT PROBIOTIC BACTERIA FOR USE IN THE TREATMENT OF A SKIN DYSFUNCTION o (57) Abstract: The invention is directed to recombinant probiotic bacteria for use in the treatment of a skin dysfunction as well as a method for treating a skin dysfunction. Recombinant probiotic bacteria for use in the treatment of a skin dysfunction The invention is directed to recombinant probiotic bacteria for use in the treatment of a skin dysfunction as well as a method for treating a skin dysfunction. Macrophages are a type of white blood cell that can be found in essentially all tissues. Macrophages play a critical role in non-specific defence and also help initiate specific defence mechanisms by recruiting other immune cells such as lymphocytes. Macrophages exist as resilient tissue-specific macrophages or are derived from circulating blood monocytes which differentiate into macrophages. Macrophages display various activation states. Two opposite activation states are known as classically activated macrophages, which are also designated as M 1- polarized macrophages, and alternatively activated macrophages, which are also designated as M2-polarized macrophages. The M 1-polarized macrophages comprise immune effector cells with an active inflammatory phenotype. M 1-polarized macrophages are characterized by the expression of high levels of pro-inflammatory cytokines, high production of reactive nitrogen and oxygen intermediates, promotion of Th1 response, and strong microbicidal and tumoricidal activity. The M2-polarized macrophages are an anti-inflammatory phenotype. M2-polarized macrophages are considered to be involved in parasite containment and promotion of tissue remodelling and tumor progression and to have immunoregulatory functions. M2-macrophages are also characterized by efficient phagocytic activity, high expression of scavenging molecules. Plasticity and flexibility are features of mononuclear phagocytes of their activation states. For example, the phenotype of M 1-polarized or M2-polarized macrophages can be reversed in vitro and in vivo. Macrophages can change, depending on the stimulus in the micro-environment, the secretion pattern of cytokine and chemokines several times. For example, human primary M 1-polarized macrophages can be repolarized by secreted factors from their own counterparts, to M2-polarized macrophages, and vice versa in vitro and in vivo. In addition to presenting a first line of defence against pathogens, mononuclear phagocytes contribute to remodelling and repair of tissue under homeostatic and damaged conditions. Moreover, macrophages are essential contributors towards the control of inflammation with M 1-polarized macrophages implicated in initiating and sustaining inflammation and M2-polarized macrophages associated with the control of chronic inflammation. For example, macrophages undergo dynamic changes during different phases of wound healing. M 1-polarized macrophages mediate tissue damage and initiate inflammatory responses. Furthermore, during the early stages of repair response of a wound, the skin infiltrating macrophages show an M2-polarized phenotype and support the formation of a highly vascularized, cellular granulation tissue. Inflammation can be characterized as acute inflammation, in settings such as sepsis, trauma, and wound healing, or as chronic inflammation, for example in diseases such as rheumatoid arthritis, ulcerative colitis, Crohn's disease, etc.. Many other diseases, such as cancer, diabetes, arteriosclerosis, Alzheimer's and obesity are also associated with deregulated inflammation. The acute inflammatory response involves a cascade of events, mediated by a large array of cells and molecules that locate invading pathogens or damaged tissue, alert and recruit other cells and molecules, eliminate the offending agents and finally restore the body to equilibrium. In sepsis and trauma, this response is accompanied by macroscopic manifestations such as fever and elevated heart rate. In other tissues, inflammation manifests as redness, swelling and pain. A feed-forward-loop of inflammation leading to damage/dysfunction leading to inflammation can result in a persistent, deregulated inflammation that promotes organ dysfunction and death. A well-regulated inflammatory response is necessary for proper tissue healing. Restoration of skin integrity and homeostasis following injuries requires a complex and dynamic interplay of epithelial and mesenchymal cells together with tissue- resident and recruited hematopoietic cells to accomplish the sequential phases of the repair response: inflammation, tissue formation, and maturation. The early stage of the repair response is dominated by the inflammatory phase, which is characterized by local activation of the innate immune system resulting in an immediate influx of neutrophiles followed by subsequent invasion of blood monocytes, which differentiate into macrophages. The mid-stage of the repair response includes the phase of tissue formation, which is characterized by the development of granulation tissue that refills the dermal wound space. Granulation tissue formation encompasses the invasion of endothelial cells resulting in angiogenesis, the influx of fibroblasts and the accumulation of additional macrophages. Deposition of provisional extracellular wound matrix facilitates cell adhesion, migration and proliferation. Furthermore, at the wound edge, complex epidermal- mesenchymal interactions stimulate keratinocyte proliferation and migration to restore the epidermal barrier. Granulation tissue formation continues until the wound space is refilled and the overlaying epidermis is restored. Upon completion of the epidermal barrier, the repair response enters the late stage, which is characterized by tissue maturation. During the phase of tissue maturation, granulation tissue transforms into scar tissue. During skin repair, the innate immune response of resident cells as well as the recruited inflammatory cells combat invading microbes, contribute to the debridement, but may also support the repair process by releasing a spectrum of growth factors. However, due to the release of proinflammatory and cytotoxic mediators, uncontrolled activity of macrophages may also be detrimental to tissue repair. An imbalanced inflammation characterized by increased numbers of macrophages is a hallmark of an attenuated repair response in human diseases leading to the formation of non-bleeding chronic wounds. Additional factors that contribute to non- bleeding chronic wounds are, for example, diabetes, venal or arterial diseases, infection, and metabolic deficiencies in old age. Various methods exist to treat chronic wounds, including the use of antibiotics for treating infections, debridement, vacuum-assisted closure, and oxygenation. Further methods include, for example, the application of growth factors. For example, becaplermin is a recombinant human platelet-derived growth factor BB. Becaplermin is sold under the trade name Regranex and is indicated for the treatment of lower extremity diabetic neuropathic ulcers that extend into the subcutaneous tissue or beyond and have an adequate blood supply. Becaplermin is further indicated as an adjunct to, and not as a substitute for, foot ulcer care practices including initial sharp debridement, pressure release and infection control. However, an increased rate of mortality secondary to malignancy was observed
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