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Connell, S.

Publications and source records attributed to Connell, S..

3 recordsLinked to original sources

EGFR-targeted and MMP-activated membranolytic peptides derived from Polybia paulista MP1 kill cancer cells specifically in vitro and reduce tumour growth in vivo

Membranolytic peptides have demonstrated potential as cancer therapeutics, although targeting to cancer cells and reducing toxicity associated with activity in normal cells remain unmet challenges. We have investigated the membranolytic peptide MP1, from Polybia paulista, in order to assess ways to reduce its non-specific toxicity and thereby improve its characteristics as a cancer therapeutic. Using a panel of human breast cell lines and cell survival assays, we show that C-terminal addition of an EGFR binding sequence, with or without a linking MMP-2 cleavage sequence, generally reduced the efficacy of the peptides relative to wildtype MP1, as determined by increases in IC50 values. Critically, cell lines that show the highest sensitivities to these fusion peptides (MDA-MB-468, MDA-MB-231) expressed the highest EGFR and/or MMP-2 levels, demonstrating that these additions direct the cytotoxic activity to cells expressing these biomarkers. MMP-2 inhibition significantly reduced the cell-killing activity of peptides containing MMP-2 cleavage sites, further demonstrating targeting to this biomarker. Fusion peptides significantly induced apoptosis and reduced survival in EGFR/MMP-2 high cancer cells, while sparing EGFR/MMP-2 low cells in the context of standard tissue culture and 3D-spheroids. Finally, systemic treatment with the EGFR and MMP-2 cleavage fusion significantly reduced tumour size in MDA-MB-468 xenograft models, confirming in vivo efficacy against cancer cells and acceptable systemic toxicity. We present this EGFR-MMP-MP1 peptide as a novel cancer therapeutic for further pre-clinical and clinical development.

cancer biology↗

Ebola's Hidden Target: Virus Transmission to and Accumulation within Skin

Ebola virus (EBOV), the causative agent of Ebola virus disease (EVD), remains one of WHOs top ten threats to global health. Infectious EBOV virions can be found on the surface of skin late during systemic infection and passed from the deceased through skin-to-skin contact. Here, we assess viral load and antigen expression in the skin of EBOV-infected non-human primates (NHP) and mouse adapted-EBOV (ma-EBOV) - infected mice and use the low containment viral model, rVSV/EBOV GP, to mechanistically define skin infection in mice. Viral RNA peaked within the skin proximal to the site of injection in EBOV-infected NHPs on day 6. In contrast, mouse skin sites distal to the site of ma-EBOV injection achieved maximal viral loads by day 3. At late times of infection, viral antigen-positive cells co-localized with markers for endothelial, stromal, and immune cells in the dermis. Epidermal cells within and surrounding hair follicles also harbored viral antigen, suggesting a potential mechanism of virus trafficking to the epidermal surface. Despite robust viral infection, distal skin sites of ma-EBOV-infected mice had low expression of proinflammatory stimulated genes. A similar cellular tropism was observed in the skin of mice infected with rVSV/EBOV GP, with discrete focal areas of intense infection. When virus was applied to the surface of gently abraded skin to remove the stratum corneum, epidermal keratinocytes were robustly infected, followed by systemic viral dissemination. To define cell surface receptors critical for virus trafficking to and replication within the skin, mice lacking the phosphatidylserine receptors were infected intraperitoneally with rVSV/EBOV GP. At day 3 of infection, skin distal to the site of infection of TIM-1 knock out (KO) mice had significantly lower levels of infectious virus than the control mice, suggesting that TIM-1 is essential for efficient distribution of virus to the skin. Our findings reveal that EBOV targets specific skin cell populations at late times of viral infection and that the host receptor TIM-1 is required for optimal viral dissemination.

microbiology↗

Multiple dermal cell types support productive infection and dynamic translocation of infectious Ebola virus to the apical surface of human skin

Ebola virus (EBOV) within the Filoviridae family causes severe human disease. At late stages of infection, EBOV virions are found on the surface of patients skin; however, the permissive cell types within the skin and how infectious virus translocates to the apical skin surfaces is not known. Here, we describe a human transwell skin explant culture model and show that EBOV infection of human skin tissues via the basal media results in a time- and dose-dependent increase in infectious virus in dermal and epidermal tissue. Infectious virus was detected on the apical epidermal surface within 3 days, indicating that the virus propagates within and traffics through the tissue. In the dermis, EBOV-infected cells were of myeloid, endothelial and fibroblast origins, whereas keratinocytes harbored virus in the epidermis. Complementary studies showed that both purified skin fibroblasts and keratinocytes supported EBOV infection ex vivo and that both cell types required the phosphatidylserine receptor, Axl, and the endosomal protein, NPC1, for virus entry. Our experimental platform identified new susceptible cell types and demonstrated dynamic trafficking of EBOV virions that resulted in infectious virus on the skin surface; findings that may explain person-to-person transmission via skin contact. TeaserUsing a human skin explant model, these studies identify and characterize skin cell populations that support Ebola virus infection.

microbiology↗