Publications
2021
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(2021) Journal of Experimental Medicine. 219, 1, e20201039. Abstract
Metastasis is the major cause of death in cancer patients. Circulating tumor cells need to migrate through the endothelial layer of blood vessels to escape the hostile circulation and establish metastases at distant organ sites. Here, we identified the membrane-bound metalloprotease ADAM17 on endothelial cells as a key driver of metastasis. We show that TNFR1-dependent tumor cell-induced endothelial cell death, tumor cell extravasation, and subsequent metastatic seeding is dependent on the activity of endothelial ADAM17. Moreover, we reveal that ADAM17-mediated TNFR1 ectodomain shedding and subsequent processing by the gamma-secretase complex is required for the induction of TNF-induced necroptosis. Consequently, genetic ablation of ADAM17 in endothelial cells as well as short-term pharmacological inhibition of ADAM17 prevents long-term metastases formation in the lung. Thus, our data identified ADAM17 as a novel essential regulator of necroptosis and as a new promising target for antimetastatic and advanced-stage cancer therapies.Metastasis is the leading cause of death in cancer patients. We discovered that proteolytic processing of endothelial TNFR1 by the metalloprotease ADAM17 is essential for metastases formation. Based on our findings, we present a novel therapeutic approach to target metastasis.
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(2021) Frontiers in Genetics. 12, 676182. Abstract
The Hippo signaling pathway has been shown to be involved in regulating cellular identity, cell/tissue size maintenance and mechanotransduction. The Hippo pathway consists of a kinase cascade which determines the nucleo-cytoplasmic localization of YAP in the cell. YAP is the effector protein in the Hippo pathway, which acts as a transcriptional cofactor for TEAD. Phosphorylation of YAP upon activation of the Hippo pathway prevents it from entering the nucleus and abrogates its function in the transcription of the target genes. In Cnidaria, the information on the regulatory roles of the Hippo pathway is virtually lacking. Here, we report the existence of a complete set of Hippo pathway core components in Hydra for the first time. By studying their phylogeny and domain organization, we report evolutionary conservation of the components of the Hippo pathway. Protein modelling suggested the conservation of YAP-TEAD interaction in Hydra. Further, we characterized the expression pattern of the homologs of yap, hippo, mob and sav in Hydra using whole-mount RNA in situ hybridization and report their possible role in stem cell maintenance. Immunofluorescence assay revealed that Hvul_YAP expressing cells occur in clusters in the body column and are excluded in the terminally differentiated regions. Actively proliferating cells marked by Ki67 exhibit YAP colocalization in their nuclei. Strikingly, a subset of these colocalized cells is actively recruited to the newly developing bud. Disruption of the YAP-TEAD interaction increased the budding rate indicating a critical role of YAP in regulating cell proliferation in Hydra. Collectively, we posit that the Hippo pathway is an essential signaling system in Hydra; its components are ubiquitously expressed in the Hydra body column and play a crucial role in Hydra tissue homeostasis.
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(2021) JACS Au. 1, 7, p. 1076-1085 Abstract
Although it is well-known that limited local mutations of enzymes, such as matrix metalloproteinases (MMPs), may change enzyme activity by orders of magnitude as well as its stability, the completely rational design of proteins is still challenging. These local changes alter the electrostatic potential and thus local electrostatic fields, which impacts the dynamics of water molecules close the protein surface. Here we show by a combined computational design, experimental, and molecular dynamics (MD) study that local mutations have not only a local but also a global effect on the solvent: In the specific case of the matrix metalloprotease MMP14, we found that the nature of local mutations, coupled with surface morphology, have the ability to influence large patches of the water hydrogen-bonding network at the protein surface, which is correlated with stability. The solvent contribution can be experimentally probed via terahertz (THz) spectroscopy, thus opening the door to the exciting perspective of rational protein design in which a systematic tuning of hydration water properties allows manipulation of protein stability and enzymatic activity.
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(2021) Cancers. 13, 7, 1679. Abstract
Matrix metalloproteases (MMPs) undergo post-translational modifications including pro-domain shedding. The activated forms of these enzymes are effective drug targets, but generating potent biological inhibitors against them remains challenging. We report the generation of anti-MMP-7 inhibitory monoclonal antibody (GSM-192), using an alternating immunization strategy with an active site mimicry antigen and the activated enzyme. Our protocol yielded highly selective anti-MMP-7 monoclonal antibody, which specifically inhibits MMP-7 s enzyme activity with high affinity (IC50 = 132 ± 10 nM). The atomic model of the MMP-7-GSM-192 Fab complex exhibited antibody binding to unique epitopes at the rim of the enzyme active site, sterically preventing entry of substrates into the catalytic cleft. In human PDAC biopsies, tissue staining with GSM-192 showed characteristic spatial distribution of activated MMP-7. Treatment with GSM-192 in vitro induced apoptosis via stabilization of cell surface Fas ligand and retarded cell migration. Co-treatment with GSM-192 and chemotherapeutics, gemcitabine and oxaliplatin elicited a synergistic effect. Our data illustrate the advantage of precisely targeting catalytic MMP-7 mediated disease specific activity.
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(2021) Journal of Dental Research. 100, 2, p. 179-186 Abstract
The periodontal ligament (PDL) plays a critical role in providing immediate response to abrupt high loads during mastication while also facilitating slow remodeling of the alveolar bone. The PDL exceptional functionality is permitted by the unique nonuniform structure of the tissue. Two distinct areas that are critical to PDL function were previously identified: the furcation and the dense collar. Despite their hypothesized functions in tooth movement and maintenance, these 2 regions have not yet been compared within the context of their native environment. Therefore, the objective of this study is to elucidate the extracellular matrix (ECM) structure, composition, and biomechanical function of the furcation and the collar regions while maintaining the 3-dimensional (3D) structure in the murine PDL. We identify significant difference between the collar and furcation regions in both structure and mechanical properties. Specifically, we observed unique longitudinal structures in the dense collar that correlate with type VI collagen and LOX, both of which are associated with increased type I collagen density and tissue stiffness and are therefore proposed to function as scaffolds for tooth stabilization. We also found that the collar region is stiffer than the furcation region and therefore suggest that the dense collar acts as a suspense structure of the tooth within the bone during physiological loading. The furcation region of the PDL contained more proteins associated with reduced stiffness and higher tissue remodeling, as well as a dual mechanical behavior, suggesting a critical function in loads transfer and remodeling of the alveolar bone. In summary, this work unravels the nonuniform nature of the PDL within the 3D structural context and establishes understanding of regional PDL function, which opens new avenues for future studies of remodeling, regeneration, and disease.
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(2021) Matrix Biology. 96, p. 47-68 Abstract
Identification of early processes leading to complex tissue pathologies, such as inflammatory bowel diseases, \u200eposes a major scientific and clinical challenge that is imperative for improved diagnosis and treatment. Most studies of inflammation onset focus on cellular processes and signaling molecules, while overlooking the environment in which they take place, the continuously remodeled extracellular matrix. In this study, we used colitis models for investigating extracellularmatrix dynamics during disease onset, while treating the matrix as a complete and defined entity. Through the analysis of matrix structure, stiffness and composition, we unexpectedly revealed that even prior to the first clinical symptoms, the colon displays its own unique extracellularmatrix signature and found specific markers of clinical potential, which were also validated in human subjects. We also show that the emergence of this pre-symptomatic matrix is mediated by subclinical infiltration of immune cells bearing remodeling enzymes. Remarkably, whether the inflammation is chronic or acute, its matrix signature converges at pre-symptomatic states. We suggest that the existence of a pre-symptomatic extracellularmatrix is general and relevant to a wide range of diseases.