Development and Validation of a Small Single-domain Antibody That Effectively Inhibits Matrix Metalloproteinase 8

Demeestere, Delphine; Dejonckheere, Eline; Steeland, Sophie; Hulpiau, Paco; Haustraete, Jurgen; Devoogdt, Nick; Wichert, Rielana; Becker‐Pauly, Christoph; Wonterghem, Elien Van; Dewaele, Sylviane; Imschoot, Griet Van; Aerts, Jeroen; Arckens, Lutgarde; Saeys, Yvan; Libert, Claude; Vandenbroucke, Roosmarijn E.

doi:10.1038/mt.2016.2

Cited by 21 publications

(13 citation statements)

References 64 publications

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“…We demonstrated that by panning with FliC, we could extract a sdAb, Abi-Se07, with a K D of 16.2 nM, within the mid-nM to high-pM range for sdAbs as reported by others (Muyldermans and Lauwereys, 1999; Demeestere et al, 2016; Sulea et al, 2018). Abi-Se07 contains a longer than average CDR3 loop (Figure 1), and could suggest that its CDR3 forms a convex paratope to access a cryptic epitope on FliC as observed with other long CDR3 sdAbs (Stijlemans et al, 2004; De Genst et al, 2006; Schmitz et al, 2013).…”

Section: Discussionsupporting

confidence: 74%

A Novel Single Domain Antibody Targeting FliC Flagellin of Salmonella enterica for Effective Inhibition of Host Cell Invasion

Huen

Zhang²,

Iwashkiw³

et al. 2019

Front. Microbiol.

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The enteric pathogen, Salmonella enterica is a major cause of human gastroenteritis globally and with increasing bacterial resistance to antibiotics, alternative solutions are urgently needed. Single domain antibodies (sdAbs), the smallest antibody fragments that retain antigen binding specificity and affinity, are derived from variable heavy-chain only fragments (VHH) of camelid heavy-chain-only immunoglobulins. SdAbs typically contain a single disulfide bond simplifying recombinant protein production in microbial systems. These factors make sdAbs ideally suited for the development of effective anti-bacterial therapeutics. To this end, we generated an anti-Salmonella VHH library from which we screened for high affinity sdAbs. We present a novel sdAb (Abi-Se07) that targets the Salmonella virulence factor, FliC, required for bacterial motility and invasion of host cells. We demonstrate that Abi-Se07 bound FliC with a KD of 16.2 ± 0.1 nM. In addition, Abi-Se07 exhibited cross-serovar binding to whole cells of S. enterica serovar Typhimurium, Heidelberg, and Hadar. Abi-Se07 significantly inhibited bacterial motility and significantly reduced S. enterica colonization in a more native environment of chicken jejunum epithelium. Taken together, we have identified a novel anti-Salmonella sdAb and discuss future efforts toward therapeutic development.

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Section: Discussionsupporting

confidence: 74%

A Novel Single Domain Antibody Targeting FliC Flagellin of Salmonella enterica for Effective Inhibition of Host Cell Invasion

Huen

Zhang²,

Iwashkiw³

et al. 2019

Front. Microbiol.

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“…EtOH treatment also increased MMP-1 and MMP-3; both have been involved in extracellular matrix disruption surrounding the RPE and the Bruch's membrane [20,31,57]. The increase in MMP-8 also corroborates the activation of the inflammation process in RPE under EtOH exposure [58].…”

Section: Discussionmentioning

confidence: 83%

Ethanol-Induced Oxidative Stress Modifies Inflammation and Angiogenesis Biomarkers in Retinal Pigment Epithelial Cells (ARPE-19): Role of CYP2E1 and its Inhibition by Antioxidants

et al. 2020

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The retinal pigment epithelium (RPE) plays a key role in retinal health, being essential for the protection against reactive oxygen species (ROS). Nevertheless, excessive oxidative stress can induce RPE dysfunction, promoting visual loss. Our aim is to clarify the possible implication of CYP2E1 in ethanol (EtOH)-induced oxidative stress in RPE alterations. Despite the increase in the levels of ROS, measured by fluorescence probes, the RPE cells exposed to the lowest EtOH concentrations were able to maintain cell survival, measured by the Cell Proliferation Kit II (XTT). However, EtOH-induced oxidative stress modified inflammation and angiogenesis biomarkers, analyzed by proteome array, ELISA, qPCR and Western blot. The highest EtOH concentration used stimulated a large increase in ROS levels, upregulating the cytochrome P450-2E1 (CYP2E1) and promoting cell death. The use of antioxidants such as N-acetylcysteine (NAC) and diallyl sulfide (DAS), which is also a CYP2E1 inhibitor, reverted cell death and oxidative stress, modulating also the upstream angiogenesis and inflammation regulators. Because oxidative stress plays a central role in most frequent ocular diseases, the results herein support the proposal that CYP2E1 upregulation could aggravate retinal degeneration, especially in those patients with high baseline oxidative stress levels due to their ocular pathology and should be considered as a risk factor.

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“…High serum or plasma levels of MMP-8 correlate to sepsis severity and fatality 39,40. Protection from endotoxin shock following LPS challenge was observed in mice treated with peptidic inhibitors of MMP-8 and MMP-9, and an MMP-8 selective nanobody 77–79. MMP-8 has been indicated to be a key mediator in the regulation of innate immunity 80.…”

Section: Discussionmentioning

confidence: 99%

Second Generation Triple-Helical Peptide Inhibitors of Matrix Metalloproteinases

et al. 2017

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The design of selective matrix metalloproteinase (MMP) inhibitors that also possess favorable solubility properties has proved to be especially challenging. A prior approach using collagen-model templates combined with transition state analogs produced a first generation of triple-helical peptide inhibitors (THPIs) that were effective in vitro against discrete members of the MMP family. These THPI constructs were also highly water-soluble. The present study sought improvements in the first generation THPIs by enhancing thermal stability and selectivity. A THPI selective for MMP-2 and MMP-9 was redesigned to incorporate non-native amino acids (Flp and mep), resulting in an increase of 18 °C in thermal stability. This THPI was effective in vivo in a mouse model of multiple sclerosis, reducing clinical severity and weight loss. Two other THPIs were developed to be more selective within the collagenolytic members of the MMP family. One of these THPIs was serendipitously more effective against MMP-8 than MT1-MMP and was utilized successfully in a mouse model of sepsis. The THPI targeting MMP-8 minimized lung damage, increased production of the anti-inflammatory cytokine IL-10, and vastly improved mouse survival.

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Development and Validation of a Small Single-domain Antibody That Effectively Inhibits Matrix Metalloproteinase 8

Cited by 21 publications

References 64 publications

A Novel Single Domain Antibody Targeting FliC Flagellin of Salmonella enterica for Effective Inhibition of Host Cell Invasion

A Novel Single Domain Antibody Targeting FliC Flagellin of Salmonella enterica for Effective Inhibition of Host Cell Invasion

Ethanol-Induced Oxidative Stress Modifies Inflammation and Angiogenesis Biomarkers in Retinal Pigment Epithelial Cells (ARPE-19): Role of CYP2E1 and its Inhibition by Antioxidants

Second Generation Triple-Helical Peptide Inhibitors of Matrix Metalloproteinases

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