Biology subjects

Lainscek, D.

Publications and source records attributed to Lainscek, D..

3 recordsLinked to original sources

bioRxiv2025

Hydroxychloroquine decouples reactogenicity from efficacy in mRNA and viral-vector gene delivery

Programmable nucleic-acid therapeutics, including lipid nanoparticle (LNP)-formulated mRNA vaccines, genome editors delivered as mRNA, and viral vectors, are transforming precision medicine but remain constrained by innate reactogenicity despite the introduction of chemical modifications into mRNA. Here, we show that mRNA/LNPs and viral vectors elicit a transient inflammatory burst that peaks [~]6 h after dosing in vivo. Co-administration or prophylaxis with hydroxychloroquine (HCQ), a clinically established 4-aminoquinoline, potently attenuated cytokine and chemokine induction across modalities, including strong responses to unmodified mRNA, without compromising therapeutic efficacy. Humoral and cellular immunity to SARS-CoV-2 Spike mRNA vaccines (BNT162b2 and non-modified mRNA/LNP) were preserved, as was live-virus neutralization. Transcriptomic profiling indicated selective dampening of TLR/cGAS-STING pathways with retention of type-I interferon elements compatible with effective vaccination. HCQ further mitigated AAV9-associated blood-brain barrier (BBB) disruption after stereotactic delivery to the mouse brain and reduced mRNA/LNP-associated hepatotoxicity and thrombocytopenia while maintaining therapeutic transgene expression and CRISPR base-editing in vivo. These findings identify HCQ as an anti-reactogenic adjunct that widens the safety window of nucleic-acid therapeutics without sacrificing performance.

immunology↗

bioRxiv2022

Designed allosteric protein logic

Regulation of the activity of proteins enables control of complex cellular processes. Allosteric regulation has been introduced individually into few natural proteins. Here, we present a generally applicable regulation of diverse proteins called INSRTR (inserted peptide structure regulator), based on inserting a short unstructured peptide into a solvent-accessible loop that retains protein function. Function of the target protein can be inactivated by the addition of a peptide that forms a rigid coiled-coil dimer. This platform enables the construction of ON/OFF protein switches, their regulation by small molecules, and Boolean logic functions with a rapid response in mammalian cells. INSRTR can be used to regulate a wide range of proteins, as demonstrated on ten members of protein families with diverse biological activities including enzymes, signaling mediators, DNA binders/transcriptional regulators, fluorescent protein, and antibodies regulating chimeric antigen receptor. INSRTR platform presents an extraordinary potential for regulating biological systems and applications. One sentence summaryAuthors have designed a widely applicable system to activate or inactivate function of diverse proteins or form Boolean logic gates based on formation of a coiled-coil dimer within protein loops and demonstrated its implementation on a range of 10 diverse proteins.

synthetic biology↗

bioRxiv2020

Immune response to vaccine candidates based on different types of nanoscaffolded RBD domain of the SARS-CoV-2 spike protein

Effective and safe vaccines against SARS-CoV-2 are highly desirable to prevent casualties and societal cost caused by Covid-19 pandemic. The receptor binding domain (RBD) of the surface-exposed spike protein of SARS-CoV-2 represents a suitable target for the induction of neutralizing antibodies upon vaccination. Small protein antigens typically induce weak immune response while particles measuring tens of nanometers are efficiently presented to B cell follicles and subsequently to follicular germinal center B cells in draining lymph nodes, where B cell proliferation and affinity maturation occurs. Here we prepared and analyzed the response to several DNA vaccines based on genetic fusions of RBD to four different scaffolding domains, namely to the foldon peptide, ferritin, lumazine synthase and {beta}-annulus peptide, presenting from 6 to 60 copies of the RBD on each particle. Scaffolding strongly augmented the immune response with production of neutralizing antibodies and T cell response including cytotoxic lymphocytes in mice upon immunization with DNA plasmids. The most potent response was observed for the 24-residue {beta}-annulus peptide scaffold that forms large soluble assemblies, that has the advantage of low immunogenicity in comparison to larger scaffolds. Our results support the advancement of this vaccine platform towards clinical trials.

immunology↗