The complement system, as integral part of innate immunity, is an important mediator between inflammatory and homeostatic processes, and participates in the clearance of debris and the defense against microbes. Moreover, its involvement in inflammatory, immune, hematological, age-related, and biomaterial-induced disorders, and its upstream position in inflammation, render complement an interesting target for therapeutic intervention. The balance between activation and regulation of alternative complement pathway (AP), and its central protein component C3, is often what tips the scale between rapid elimination of danger and inapt attack of host cells. Furthermore, human pathogens employ AP inhibition as immune escape strategy. Molecular insight into the processes that shape the activity of C3 therefore has important implications for understanding the role of complement in health, disease and infection, and for the development of complement therapeutics. Based on accomplishments that produced seminal structures and revealed mechanisms of C3-mediated complement activity, this proposal aims at the molecular description of components/events that shape initiation and regulation of the AP and opsonic signaling. The dynamic conversion of C3 to its hydrolytically activated form C3(H2O) will be studied by a combination of structural, biochemical and biophysical methods to reveal its structure and reactivity towards activators and regulators. Careful comparison with C3b as the driving molecule of AP amplification will define determinants of AP activation and provide insight into triggering processes that contribute to inflammatory complications caused by biomaterials or blood oxygenation. Surface opsonization is a direct result of AP amplification and drives phagocytosis and stimulation of adaptive immune responses. In contrast to C3b, the molecular features of the versatile opsonin iC3b that binds to five complement receptors (CR) are still insufficiently described. Structural characterization of iC3b and exploration of its CR recognition pattern will provide important information about the factors that define selectivity and activity ~ the integrin receptor CR3 is of particular interest due to its importance in innate immunity and potential as drug target. Novel cellular assays will extend the study of opsonic signaling at the cellular level and may emerge as tools for drug screening and phagocytic monitoring. Finally, key determinants of physiological AP regulation on host cells will be defined by structural and functional studies of 'regulators of complement activation'(RCA) and microbial mimics thereof. Due to their structural modularity and high AP-directed potency, RCA are considered promising templates for complement inhibitors. Supported on the insight on RCA activity and opsonin specificity as elucidated in this proposal, surface-targeted RCA-based inhibitors will be engineered and evaluated in human and mouse models of complement-mediated disease. One candidate already showed high promise in a model of paroxysmal nocturnal hemoglobinuria, in which it largely surpassed other inhibitors. The proposed studies therefore have high impact for immunology, biomedicine, and drug discovery.

Public Health Relevance

Here we provide molecular insight into activation, regulation and signaling of a central pillar of innate immunity, the human complement system, by resolving the structures and functional mechanisms of several key proteins with direct implications for immunology and biomedicine. In view of the strong involvement of complement in inflammatory, immune, and age-related diseases, a particular focus is set on resolving molecular disease associations and the development of a novel class of potent complement inhibitors based on a promising candidate protein.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI030040-16
Application #
8578765
Study Section
Innate Immunity and Inflammation Study Section (III)
Program Officer
Leitner, Wolfgang W
Project Start
1992-12-01
Project End
2017-08-31
Budget Start
2013-09-01
Budget End
2014-08-31
Support Year
16
Fiscal Year
2013
Total Cost
$366,753
Indirect Cost
$128,023
Name
University of Pennsylvania
Department
Pathology
Type
Schools of Medicine
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Schmidt, Christoph Q; Harder, Markus J; Nichols, Eva-Maria et al. (2016) Selectivity of C3-opsonin targeted complement inhibitors: A distinct advantage in the protection of erythrocytes from paroxysmal nocturnal hemoglobinuria patients. Immunobiology 221:503-11
Mastellos, Dimitrios C; Reis, Edimara S; Yancopoulou, Despina et al. (2016) From orphan drugs to adopted therapies: Advancing C3-targeted intervention to the clinical stage. Immunobiology 221:1046-57
Hajishengallis, George; Lambris, John D (2016) More than complementing Tolls: complement-Toll-like receptor synergy and crosstalk in innate immunity and inflammation. Immunol Rev 274:233-244
Lindorfer, Margaret A; Cook, Erika M; Reis, Edimara S et al. (2016) Compstatin Cp40 blocks hematin-mediated deposition of C3b fragments on erythrocytes: Implications for treatment of malarial anemia. Clin Immunol 171:32-35
Maekawa, Tomoki; Briones, Ruel A; Resuello, Ranillo R G et al. (2016) Inhibition of pre-existing natural periodontitis in non-human primates by a locally administered peptide inhibitor of complement C3. J Clin Periodontol 43:238-49
Hajishengallis, George; Hajishengallis, Evlambia; Kajikawa, Tetsuhiro et al. (2016) Complement inhibition in pre-clinical models of periodontitis and prospects for clinical application. Semin Immunol 28:285-91
Ricklin, Daniel; Reis, Edimara S; Lambris, John D (2016) Complement in disease: a defence system turning offensive. Nat Rev Nephrol 12:383-401
Wang, Junxiang; Wang, Lu; Xiang, Ying et al. (2016) Using an in vitro xenoantibody-mediated complement-dependent cytotoxicity model to evaluate the complement inhibitory activity of the peptidic C3 inhibitor Cp40. Clin Immunol 162:37-44
Mastellos, D C; Ricklin, D; Hajishengallis, E et al. (2016) Complement therapeutics in inflammatory diseases: promising drug candidates for C3-targeted intervention. Mol Oral Microbiol 31:3-17
Hajishengallis, George; Krauss, Jennifer L; Jotwani, Ravi et al. (2016) Differential capacity for complement receptor-mediated immune evasion by Porphyromonas gingivalis depending on the type of innate leukocyte. Mol Oral Microbiol :

Showing the most recent 10 out of 159 publications