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Vandeloo, M.

Publications and source records attributed to Vandeloo, M..

3 recordsLinked to original sources

What comes after de novo? Automated lead optimization of proteins with CRADLE-1

Lead optimization remains the longest and most expensive step in pre-clinical drug discovery, typically consuming 12-36 months whilst costing $5M-$15M per candidate. We introduce O_SCPLOWCRADLEC_SCPLOWO_SCPCAP-1C_SCPCAP, an automated framework for protein engineering. While O_SCPLOWCRADLEC_SCPLOWO_SCPCAP-1C_SCPCAP supports the full process of drug discovery and industrial protein engineering pipelines, including hit identification and de novo binder design, this work focuses on its application to multi-property lead optimization across protein modalities (VHHs, scFvs, IgGs, peptides, enzymes, CRISPR systems, vaccines). We show it is 4-7x faster than rational design, as measured by the number of wet lab rounds required. We provide in-vitro validation across all of the above modalities, typically optimizing multiple properties simultaneously (single and polyspecific binding down to picomolar, activity, thermostability,...). Technically, O_SCPLOWCRADLEC_SCPLOWO_SCPCAP-1C_SCPCAP starts with pre-trained foundation protein language models (PLMs), which are fine-tuned in unsupervised fashion on evolutionary neighborhoods, in supervised fashion using lab-in-the-loop data, and then deployed in a multi-model workflow. Of additional interest, we find that (a) the end-to-end system may be run in automated fashion; (b) wet lab data may be consumed in black box fashion without knowledge of the underlying biochemical mechanisms; (c) structural data may largely be superseded by sequence-function pairs.

bioengineering↗

Integrated Metabolic Complex Genetic Interaction 1 Network of Chromosome 4p Loss in Basal Breast Cancer

Basal breast cancer subtype is enriched for triple-negative breast cancer (TNBC) and exhibits a recurrent large chromosomal deletion in chromosome 4p (chr4p). Chr4p loss is associated with poor survival, evolves early in tumorigenesis and confers on cells a proliferative state. Here, we map the integrated metabolic complex genetic interaction network of chr4p in basal breast cancer to identify targetable vulnerabilities. Differential gene expression analysis of patient derived xenografts and cancer cell models revealed that chr4p loss is associated with changes in cellular energetics and reduction/oxidation balance. Analysis of DepMap pooled genome-wide CRISPR-Cas9 screens identified complex genetic interactions specific to chr4p deletion in basal breast cancer cell models. Functional assays revealed that chr4p loss is associated with disrupted mitochondrial respiratory function and reduced glycolytic capacity, suggesting metabolic rewiring. Increased reactive oxygen species and lipid peroxidation compromised antioxidant defense mechanisms. Ultimately, this study sheds light on targeted therapies for basal breast cancer harboring large chromosomal deletions.

genomics↗

Rapid, scalable, combinatorial genome engineering by Marker-less Enrichment and Recombination of Genetically Engineered loci (MERGE)

Large-scale genome engineering in yeast is feasible primarily due to prodigious homology-directed DNA repair (HDR), a plethora of genetic tools, and simple conversion between haploid and diploid forms. However, a major challenge to rationally building multi-gene processes in yeast arises due to the combinatorics of combining all of the individual edits into the same strain. Here, we present an approach for scalable, precise, multi-site genome editing that combines all edits into a single strain without the need for selection markers by using CRISPR-Cas9 and gene drives. First, we show that engineered loci become resistant to the corresponding CRISPR reagent, allowing the enrichment of distinct genotypes. Next, we demonstrate a highly efficient gene drive that selectively eliminates specific loci by integrating CRISPR-Cas9 mediated Double-Strand Break (DSB) generation and homology-directed recombination with yeast sexual assortment. The method enables Marker-less Enrichment and Recombination of Genetically Engineered loci (MERGE) in yeast. We show that MERGE converts single heterologous yeast loci to homozygous loci at [~]100% efficiency, independent of chromosomal location. Furthermore, MERGE is equally efficient at converting and combining loci, thus identifying viable intermediate genotypes. Finally, we establish the feasibility of MERGE by engineering a fungal carotenoid biosynthesis pathway and most of the human proteasome core into yeast. MERGE, therefore, lays the foundation for marker-less, highly efficient, and scalable combinatorial genome editing in yeast.

synthetic biology↗