Arzneistoffe aus Bakterien: YopM als bakterielles, anti-inflammatorisches, Zell-penetrierendes Peptid für die topische Behandlung von Psoriasis

• Antragstellerinnen / Antragsteller: Professorin Dr. Karin Loser; Privatdozent Dr. Christian Rüter
• Fachliche Zuordnung: Dermatologie ; Immunologie
• Förderung: Förderung von 2014 bis 2019
• Projektkennung: Deutsche Forschungsgemeinschaft (DFG) - Projektnummer 258026263

• Erstellungsjahr: 2019

Zusammenfassung der Projektergebnisse

Psoriasis is a chronic inflammatory skin disease affecting 2-4 % of the human population. The disease is characterized by red, itchy and scaly plaques and markedly impairs the patient’s quality of life. Albeit several efficient treatment options are available the disease cannot be completely cures, which means that only the inflammatory symptoms can be adequately treated. More effective treatments are often associated with increasing toxicity and/or costs. Therefore, the development of new local, potentially low cost therapeutics is needed. The Yersinia outer protein M (YopM) was the first bacterial effector protein discovered to have the characteristics of a cell-penetrating peptide (CPP), meaning it is able to autonomously cross eukaryotic cell membranes independent of the Type 3 secretion system (T3SS). Thereafter, it can be detected in the cytoplasm as well as in the nucleus of the host cell. It has been demonstrated that, once inside the cell, recombinant YopM (rYopM) acts anti-inflammatory by downregulating the expression of pro-inflammatory cytokines, like TNF-α or IL-1β. Hence, rYopM might be a promising molecule for further investigations in the context of inflammatory skin diseases, like psoriasis. In this research project the suitability of rYopM as a therapeutic drug was tested in the the mouse model of imiquimod (IMQ)-induced psoriasis-like skin disease. We were able to demonstrate that the subcutaneous injection of rYopM as well as the topical application as a cream ameliorated ongoing inflammation. Interestingly, treatment with rYopM did not have a major effect on the T cell compartment but the compound led to a reduced proliferative activity of keratinocytes. Of note, keratinocyte hyperproliferation and misguided differentiation is a histological hallmark of psoriatic plaques. Further, rYopM reduced the protein levels of TNF-α, IL-1β and the neutrophil-attracting chemokines CXCL1 and CXCL5. By negatively regulating the levels of these proteins rYopM treatment led to a downregulated infiltration of neutrophils. To better characterize the functional groups of the protein, truncated versions were generated and tested for their anti-inflammatory efficacy in cell culture and in a mouse model. Notably, a minimal version wherein only the two N-terminal α-helices (essential for translocation) and the first three leucine-rich repeats (LRRs) are present has a comparable effect as compared to full-length rYopM on keratinocyte proliferation and neutrophil infiltration. Hence these data might suggest rYopM and its minimal variants as highly promising targets for further analysis and the development towards a potential clinical application in the context of inflammatory skin diseases including psoriasis.

Projektbezogene Publikationen (Auswahl)

• (2013)„Cell-penetrating bacterial E3-ubiqitin-ligases for use in immunetherapy”. PCT-Patent Application ref.: 13 185 412.7. Australian and US Patent granted in 2019
  Rüter C., Lubos M. and Schmidt M.A.
  
• (2015) Cutaneous RANK-RANKL Signaling Upregulates CD8-Mediated Antiviral Immunity during Herpes simplex Virus Infection by Preventing Virus-Induced Langerhans Cell Apoptosis. J Invest Dermatol 135(11):2676-2687
  Klenner L, Hafezi W, Clausen BE, Lorentzen EU, Luger TA, Beissert S, Kühn JE, Loser K
  
• NDP-MSH for the treatment of inflammatory and/or neurodegenerative disorders of the CNS (PCT/EP2014/066816; international filing date August 5th, 2014; publication February 2nd, 2015 [WO 2015/018827])
  Loser K, Luger TA
  
• (2016) Melanocortin-1 receptor activation is neuroprotective in mouse models of neuroinflammatory disease. Sci Transl Med 8(362):362ra146
  Mykicki N, Herrmann AM, Schwab N, Deenen R, Sparwasser T, Limmer A, Wachsmuth L, Klotz L, Köhrer K, Faber C, Wiendl H, Luger TA, Meuth SG, Loser K
  
• (2016) Sensitive skin. J Eur Acad Dermatol Venereol, Suppl 1:2-8
  Misery L, Loser K, Ständer S
  
• (2017) Cell-Penetrating Bacterial Effector Proteins: Better Tools than Targets. Trends in Biotechnology. 35(2):109-120
  Rüter C., and Schmidt M.A.
  
• (2017) Immunomodulatory Yersinia outer proteins (Yops) -useful tools for bacteria and humans alike. Virulence 8(7):1124-1147
  Grabowski B., Schmidt M.A. and Rüter C.
  
• (2017) The tripeptide KdPT ameliorates ongoing psoriasis-like skin inflammation in murine and human skin. Exp Dermatol 26(4):328-334
  Mykicki N, Klenner L, Baumann C, Auriemma M, Sternemann C, Soeberdt M, Elliott GR, Abels C, Luger TA, Loser K
  
• (2018) All in-Multiple parallel strategies for intracellular delivery by bacterial pathogens. Int J Med Microbiol. 308(7):872-881
  Rüter C., Lubos M-L., Norkowski S., Schmidt M.A.
  
• (2018) Novel insights into the pathogenesis of psoriasis. Clin Immunol 186:43-45
  Luger TA, Loser K
  
• (2019) Activation of human vascular endothelium in melanoma metastases induces ICAM-1 and E- selectin expression and results in increased infiltration with effector lymphocytes. Exp Dermatol.
  Weishaupt C, Steinert M, Brunner G, Schulze HJ, Fuhlbrigge RC, Goerge T, Loser K
  
• (2019) Development of Novel Quinoxaline-Based κ-Opioid Receptor Agonists for the Treatment of Neuroinflammation. J Med Chem 62(2):893-907
  Tangherlini G, Kalinin DV, Schepmann D, Che T, Mykicki N, Ständer S, Loser K, Wünsch B