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Beyond envenomation: functional dissection of Podalia orsilochus venom uncovers a procoagulant protein and additional toxic activities

ABSTRACT: Envenomation by urticating caterpillars of the family Megalopygidae is an underestimated public health issue in South America, with Podalia orsilochus being the main species implicated in cases reported in northeastern Argentina. This study aimed to characterize the toxicological properties of its venom, focusing on hematological and hemostatic effects. Venom extracts were analyzed using electrophoresis, zymography, hemolysis and coagulation assays, and chromatographic fractionation followed by immunodetection. Electrophoretic profiles revealed predominant proteins in the 10–37 kDa range, with several gelatinolytic bands detected by zymography. The venom exhibited thrombin-like and dose-dependent procoagulant activity, although weaker than that of Lonomia obliqua. A mild, direct hemolytic effect on human erythrocytes was observed, associated with degradation of Band 3, a major membrane protein. A 33-kDa protein was purified and shown to induce procoagulant activity while cross-reacting with antibodies against Losac, a Factor X-activating protein from L. obliqua venom, indicating the presence of Losac-like toxins in P. orsilochus. In conclusion, P. orsilochus venom contains components with proteolytic, hemolytic, and coagulotoxic activities, including a Losac-like protein, expanding current knowledge of lepidopteran venoms and their potential impact on human health.

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ORGANISM(S): Megalopygidae Sp. Bold:aaa3544

SUBMITTER: Maria Pia Valacco

LAB HEAD: Maria Elisa Paichoto

PROVIDER: PXD074932 | Pride | 2026-03-30

REPOSITORIES: Pride

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Project description:Regenerative therapy employing intestinal stem cells (ISCs) holds a great promise for reliable epithelial restoration. However, achieving this goal requires scalable and robust, but fully defined culture platforms that eliminate the use of xenogeneic components while preserving stem cell function. Most existing systems usually rely on the use of Matrigel, complicating clinical translation and limiting mechanistic understanding of stem cell-material interactions. In this study, a poly(ethyleneglycoldimethacrylate) (pEGDMA)-based synthetic culture surface is precisely refined through N2 plasma treatment thereon, enabling tailored modulation of surface properties. This modification enhances wettability of the polymer surface and introduces N-containing functional groups, resulting in the PoLymer-coated Ultra-stable Surface (PLUS) that significantly improves ISC attachment under xenogeneic-free conditions. Remarkably, PLUS maintains its ISC-supportive function even after three years of ambient storage. Gene expression analysis and comparative proteomic profiling clearly reveal the upregulation of factors involved in actin dynamics and cytoskeletal reorganization, indicating a structural basis for enhanced colony expansion. Consistently, colonies on PLUS exhibit a broader migratory range and migrate 1.8-fold faster during random movement than those on pristine pEGDMA. Functional validation using small-molecule perturbation assays confirms the involvement of cytoskeletal remodeling by exquisitely mediating ISC-substrate interaction. Furthermore, PLUS enables progressive wound closure via dynamic migration by up to 46.7% within 144 h through actin-dependent mechanisms, supporting the facilitated epithelium regeneration. By providing a long-term reliable bioactive surface that sustains stemness and regenerative capacity of ISCs, PLUS engages cytoskeletal machinery as a central mediator of ISC-substrate interaction, in part by promoting actin-dependent cytoskeletal reorganization, positioning it as a scalable, translational platform for intestinal stem cell-based regenerative medicine.

Project description:Background The generalist dipteran pupal parasitoid Nasonia vitripennis injects 79 venom peptides into the host before egg laying. This venom induces several important changes in the host, including developmental arrest, immunosuppression, and alterations to normal metabolism. It is hoped that diverse and potent bioactivities of N. vitripennis venom provide an opportunity for the design of novel acting drugs. However, currently very little is known about the individual functions of N. vitripennis venom peptides and less than half can be bioinformatically annotated. The paucity of annotation information complicates the design of studies that seek to better understand the potential mechanisms underlying the envenomation response. Although the RNA interference system of N. vitripennis provides an opportunity to functionally characterise venom encoding genes, with 79 candidates this represents a daunting task. For this reason we were interested in determining the expression levels of venom encoding genes in the venom gland, such that this information could be used to rank candidate venoms. To do this we carried out deep sequencing of the transcriptome of the venom gland and neighbouring ovary tissue and used RNA-seq to measure expression from the 79 venom encoding genes. The generation of a specific venom gland transcriptome dataset also provides further opportunities to investigate novel features of this highly specialised organ. Results High throughput sequencing and RNA-seq revealed that the highest expressed venom encoding gene in the venom gland was a serine protease called Nasvi2EG007167, which has previously been implicated in the apoptotic activity of N. vitripennis venom. As expected the RNA-seq confirmed that the N. vitripennis venom encoding genes are almost exclusively expressed in the venom gland relative to the neighbouring ovary tissue. Novel peptides appear to perform key roles in N. vitripennis venom function as only four of the highest 15 expressed venom encoding genes are bioinformatically annotationed. The high throughput sequencing data also provided evidence for the existence of an additional 471 novel genes in the Nasonia genome that are expressed in the venom gland and ovary. Finally, metagenomic analysis of venom gland transcripts identified viral transcripts that may play an important part in the N. vitripennis venom function. Conclusions The expression level information provided here for the 79 venom encoding genes provides an unbiased dataset that can be used by the N. vitripennis community to identify high value candidates for further functional characterisation. These candidates represent bioactive peptides that have value in drug development pipelines.

			Action	DRS
	Contaminantes.fasta	Fasta
	MEP32.raw	Raw
	MEP32a.msf	Msf
	MEP32b.msf	Msf
	checksum.txt	Txt

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Beyond envenomation: functional dissection of Podalia orsilochus venom uncovers a procoagulant protein and additional toxic activities

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