Development of Procathepsin L (pCTS-L)-Inhibiting Lanosterol-Carrying Liposome Nanoparticles to Treat Lethal Sepsis

The pathogenesis of microbial infections and sepsis is partly attributable to dysregulated innate immune responses propagated by late-acting proinflammatory mediators such as procathepsin L (pCTS-L). It was previously not known whether any natural product could inhibit pCTS-L-mediated inflammation or could be strategically developed into a potential sepsis therapy. Here, we report that systemic screening of a NatProduct Collection of 800 natural products led to the identification of a lipophilic sterol, lanosterol (LAN), as a selective inhibitor of pCTS-L-induced production of cytokines [e.g., Tumor Necrosis Factor (TNF) and Interleukin-6 (IL-6)] and chemokines [e.g., Monocyte Chemoattractant Protein-1 (MCP-1) and Epithelial Neutrophil-Activating Peptide (ENA-78)] in innate immune cells. To improve its bioavailability, we generated LAN-carrying liposome nanoparticles and found that these LAN-containing liposomes (LAN-L) similarly inhibited pCTS-L-induced production of several chemokines [e.g., MCP-1, Regulated upon Activation, Normal T Cell Expressed and Presumably Secreted (RANTES) and Macrophage Inflammatory Protein-2 (MIP-2)] in human blood mononuclear cells (PBMCs). In vivo, these LAN-carrying liposomes effectively rescued mice from lethal sepsis even when the first dose was given at 24 h post the onset of this disease. This protection was associated with a significant attenuation of sepsis-induced tissue injury and systemic accumulation of serval surrogate biomarkers [e.g., IL-6, Keratinocyte-derived Chemokine (KC), and Soluble Tumor Necrosis Factor Receptor I (sTNFRI)]. These findings support an exciting possibility to develop liposome nanoparticles carrying anti-inflammatory sterols as potential therapies for human sepsis and other inflammatory diseases.

[1]  K. Tracey,et al.  Identification of procathepsin L (pCTS-L)–neutralizing monoclonal antibodies to treat potentially lethal sepsis , 2023, Science advances.

[2]  David L. Williams,et al.  Lactate promotes endothelial-to-mesenchymal transition via Snail1 lactylation after myocardial infarction , 2023, Science advances.

[3]  K. Raza,et al.  Proliposome-Based Nanostrategies: Challenges and Development as Drug Delivery Systems , 2022, AAPS PharmSciTech.

[4]  Abdelwahab Omri,et al.  Lipid-Based Drug Delivery Systems for Diseases Managements , 2022, Biomedicines.

[5]  David L. Williams,et al.  Lactate induces vascular permeability via disruption of VE-cadherin in endothelial cells during sepsis , 2022, Science advances.

[6]  Neha Dhiman,et al.  A drift on liposomes to proliposomes: recent advances and promising approaches , 2022, Journal of liposome research.

[7]  K. Tracey,et al.  Human Dermcidin Protects Mice Against Hepatic Ischemia-Reperfusion–Induced Local and Remote Inflammatory Injury , 2022, Frontiers in Immunology.

[8]  David S. Fischer,et al.  Single-cell meta-analysis of SARS-CoV-2 entry genes across tissues and demographics , 2021, Nature Medicine.

[9]  D. Heffernan,et al.  Emerging therapeutic targets for sepsis , 2021, Expert opinion on therapeutic targets.

[10]  Yutian Li,et al.  Tissue-Resident Macrophages in the Control of Infection and Resolution of Inflammation. , 2020, Shock.

[11]  K. Tracey,et al.  Identification of tetranectin-targeting monoclonal antibodies to treat potentially lethal sepsis , 2020, Science Translational Medicine.

[12]  T. Buchman,et al.  Sepsis Among Medicare Beneficiaries: 1. The Burdens of Sepsis, 2012–2018* , 2020, Critical care medicine.

[13]  Niranjan Kissoon,et al.  Global, regional, and national sepsis incidence and mortality, 1990–2017: analysis for the Global Burden of Disease Study , 2020, The Lancet.

[14]  C. Deutschman,et al.  Buprenorphine Markedly Elevates a Panel of Surrogate Markers in a Murine Model of Sepsis , 2019, Shock.

[15]  K. Tracey,et al.  Enhanced Macrophage Pannexin 1 Expression and Hemichannel Activation Exacerbates Lethal Experimental Sepsis , 2019, Scientific Reports.

[16]  Minoru Yoshida.  Recent advances in target identification of bioactive natural products , 2019, Bioscience, biotechnology, and biochemistry.

[17]  K. Tracey,et al.  Connexin 43 Hemichannel as a Novel Mediator of Sterile and Infectious Inflammatory Diseases , 2018, Scientific Reports.

[18]  M. A. Lasunción,et al.  Lanosterol Modulates TLR4-Mediated Innate Immune Responses in Macrophages. , 2017, Cell reports.

[19]  Huan Yang,et al.  Cathepsin L Promotes Vascular Intimal Hyperplasia after Arterial Injury , 2017, Molecular medicine.

[20]  Y. Li,et al.  Cathepsin L activity correlates with proteinuria in chronic kidney disease in humans , 2017, International Urology and Nephrology.

[21]  Mingzi M. Zhang,et al.  Using natural products for drug discovery: the impact of the genomics era , 2017, Expert opinion on drug discovery.

[22]  A. Cilla,et al.  Impact of Lipid Components and Emulsifiers on Plant Sterols Bioaccessibility from Milk-Based Fruit Beverages. , 2016, Journal of agricultural and food chemistry.

[23]  Susan Hua,et al.  Advances and Challenges of Liposome Assisted Drug Delivery , 2015, Front. Pharmacol..

[24]  Ming-Wei Lin,et al.  Euphol from Euphorbia tirucalli Negatively Modulates TGF-β Responsiveness via TGF-β Receptor Segregation inside Membrane Rafts , 2015, PloS one.

[25]  D. Vodnar,et al.  Antioxidant, Antimicrobial Effects and Phenolic Profile of Lycium barbarum L. Flowers , 2015, Molecules.

[26]  Ruben Abagyan,et al.  Lanosterol reverses protein aggregation in cataracts , 2015, Nature.

[27]  V. Nekkanti,et al.  Proliposomes for oral delivery: progress and challenges. , 2015, Current pharmaceutical biotechnology.

[28]  Haichao Wang,et al.  PKM2 Regulates the Warburg Effect and Promotes HMGB1 Release in Sepsis , 2014, Nature Communications.

[29]  Haichao Wang,et al.  Carbenoxolone Blocks Endotoxin-Induced Protein Kinase R (PKR) Activation and High Mobility Group Box 1 (HMGB1) Release , 2013, Molecular Medicine.

[30]  Soodabeh Davaran,et al.  Liposome: classification, preparation, and applications , 2013, Nanoscale Research Letters.

[31]  G. F. Passos,et al.  The role of PKC/ERK1/2 signaling in the anti-inflammatory effect of tetracyclic triterpene euphol on TPA-induced skin inflammation in mice. , 2013, European journal of pharmacology.

[32]  J. Calixto,et al.  Euphol, a tetracyclic triterpene produces antinociceptive effects in inflammatory and neuropathic pain: The involvement of cannabinoid system , 2012, Neuropharmacology.

[33]  C. Peters,et al.  Decreased arthritis severity in cathepsin L-deficient mice is attributed to an impaired T helper cell compartment , 2012, Inflammation Research.

[34]  J. Calixto,et al.  Euphol prevents experimental autoimmune encephalomyelitis in mice: evidence for the underlying mechanisms. , 2012, Biochemical pharmacology.

[35]  H. Mansour,et al.  Therapeutic Liposomal Dry Powder Inhalation Aerosols for Targeted Lung Delivery , 2012, Lung.

[36]  J. Calixto,et al.  Preventive and Therapeutic Euphol Treatment Attenuates Experimental Colitis in Mice , 2011, PloS one.

[37]  K. Tracey,et al.  EGCG stimulates autophagy and reduces cytoplasmic HMGB1 levels in endotoxin-stimulated macrophages. , 2011, Biochemical pharmacology.

[38]  J. Tschopp,et al.  Colitis induced in mice with dextran sulfate sodium (DSS) is mediated by the NLRP3 inflammasome , 2010, Gut.

[39]  C. Peters,et al.  Cathepsin L inactivates human trypsinogen, whereas cathepsin L-deletion reduces the severity of pancreatitis in mice. , 2010, Gastroenterology.

[40]  Huan Yang,et al.  A Major Ingredient of Green Tea Rescues Mice from Lethal Sepsis Partly by Inhibiting HMGB1 , 2007, PloS one.

[41]  Fang-Yu Chen,et al.  The condensing effect of cholesterol in lipid bilayers. , 2007, Biophysical journal.

[42]  Christoph Peters,et al.  Cathepsin L Deficiency Reduces Diet-Induced Atherosclerosis in Low-Density Lipoprotein Receptor-Knockout Mice , 2007, Circulation.

[43]  Javed Siddiqui,et al.  Circulating Cytokine/Inhibitor Profiles Reshape the Understanding of the SIRS/CARS Continuum in Sepsis and Predict Mortality1 , 2006, The Journal of Immunology.

[44]  K. Tracey,et al.  Role of HMGB1 in apoptosis-mediated sepsis lethality , 2006, The Journal of Experimental Medicine.

[45]  T. Starc,et al.  Intestinal cholesterol absorption inhibitor ezetimibe added to cholestyramine for sitosterolemia and xanthomatosis. , 2006, Gastroenterology.

[46]  S. Krueger,et al.  Bilayer thickness and thermal response of dimyristoylphosphatidylcholine unilamellar vesicles containing cholesterol, ergosterol and lanosterol: a small-angle neutron scattering study. , 2005, Biochimica et biophysica acta.

[47]  K. Tracey,et al.  Reversing established sepsis with antagonists of endogenous high-mobility group box 1 , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[48]  R. Winter,et al.  Differential Properties of the Sterols Cholesterol, Ergosterol, β-Sitosterol, trans-7-Dehydrocholesterol, Stigmasterol and Lanosterol on DPPC Bilayer Order , 2003 .

[49]  A. Harken,et al.  TLR4 mediates LPS-induced HO-1 expression in mouse liver: Role of TNF-α and IL-1β , 2003 .

[50]  M. Bloom,et al.  From lanosterol to cholesterol: structural evolution and differential effects on lipid bilayers. , 2002, Biophysical journal.

[51]  K. Tracey,et al.  HMG-1 as a late mediator of endotoxin lethality in mice. , 1999, Science.

[52]  P. Ricciardi-Castagnoli,et al.  Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene. , 1998, Science.

[53]  K. Tracey,et al.  Spermine Inhibits Proinflammatory Cytokine Synthesis in Human Mononuclear Cells: A Counterregulatory Mechanism that Restrains the Immune Response , 1997, The Journal of experimental medicine.

[54]  C. Dinarello,et al.  Biologic basis for interleukin-1 in disease. , 1996, Blood.

[55]  Kevin J. Tracey,et al.  Anti-cachectin/TNF monoclonal antibodies prevent septic shock during lethal bacteraemia , 1987, Nature.