bioRxiv Science⌕ Search

Biology subjects

Rotenberg, N.

Publications and source records attributed to Rotenberg, N..

2 recordsLinked to original sources

A trustworthy semantic knowledge network linking data-driven cell phenotypes to diseases and drugs

Genome-wide association and other genetic studies have identified genes involved in disease pathogenesis, and their encoded proteins have provided therapeutic targets. However, little is known about which cells are the functional home of these disease-associated genes and proteins. Single-cell technologies are enabling high-resolution profiling of cell phenotypes and revealing cellular contexts of disease, yet much of the resulting knowledge remains embedded in individual datasets and scientific literature. To address these gaps, the National Library of Medicine has developed Cell Knowledge Network (NLM-CKN, https://nlm-ckn.org). Reproducible computational workflows have generated cell phenotype knowledge for 2,585 cell sets and 5,139 selectively expressed genes from 88 datasets covering 10 human organs. Mapping of these cell sets to cell types grounds Cell Ontology terms in experimental evidence. This experimental knowledge is integrated with external knowledge of anatomical and cell-type hierarchies, gene-disease associations, and therapeutic interventions. A semantic schema defines node types and relations, enabling knowledge to be transformed into triple assertions and loaded into an ArangoDB database with provenance and quantitative evidence. The NLM-CKN interface supports graphical exploration of the cellular constituents of healthy human tissues. Finally, topological analyses generate insights into cellular contributions to disease pathogenesis, cellular contexts for therapeutic intervention, and drug-repurposing hypotheses.

bioinformatics↗

Improving Circulation Half-Life of Therapeutic Candidate N-TIMP2 by Unfolded Peptide Extension

Matrix Metalloproteinases (MMPs) are drivers of many diseases including cancer and are established targets for drug development. Tissue inhibitors of metalloproteinases (TIMPs) are human proteins that inhibit MMPs and are being pursued for the development of anti-MMP therapeutics. TIMPs possess many attractive properties of a drug candidate, such as complete MMP inhibition, low toxicity and immunogenicity, high tissue permeability and others. A major challenge with TIMPs, however, is their formulation and delivery, as these proteins are quickly cleared from the bloodstream due to their small size. In this study, we explore a new method for plasma half-life extension for the N-terminal domain of TIMP2 (N-TIMP2) through appending it with a long intrinsically unfolded tail containing a random combination of Pro, Ala, and Thr (PATylation). We design, produce and explore two PATylated N-TIMP2 constructs with a tail length of 100- and 200-amino acids (N-TIMP2-PAT100 and N-TIMP2-PAT200, respectively). We demonstrate that both PATylated N-TIMP2 constructs possess apparent higher molecular weights compared to the wild-type protein and retain high inhibitory activity against MMP-9. Furthermore, when injected into mice, N-TIMP2-PAT200 exhibited a significant increase in plasma half-life compared to the non-PATylated variant, enhancing the therapeutic potential of the protein. Thus, we establish that PATylation could be successfully applied to TIMP-based therapeutics and offers distinct advantages as an approach for half-life extension, such as fully genetic encoding of the gene construct, mono-dispersion, and biodegradability. Furthermore, PATylation could be easily applied to N-TIMP2 variants engineered to possess high affinity and selectivity toward individual MMP family members, thus creating attractive candidates for drug development against MMP-related diseases.

biochemistry↗