Targeting antitumoral proteins to breast cancer by local administration of functional inclusion bodies

Altres ajuts de l'Instituto de Salud Carlos III: PI15/00272, PI1702242FIS i #PI16/01224Two structurally and functionally unrelated proteins, namely Omomyc and p31, are engineered as CD44-targeted inclusion bodies produced in recombinant bacteria. In this unusual particulate form, both types of protein materials selectively penetrate and kill CD44 tumor cells in culture, and upon local administration, promote destruction of tumoral tissue in orthotropic mouse models of human breast cancer. These findings support the concept of bacterial inclusion bodies as versatile protein materials suitable for application in chronic diseases that, like cancer, can benefit from a local slow release of therapeutic protein

Androgen receptor condensates as drug targets

Transcription factors are among the most attractive therapeutic targets, but are considered largely undruggable. Here we provide evidence that small molecule-mediated partitioning of the androgen receptor, an oncogenic transcription factor, into phase-separated condensates has therapeutic effect in prostate cancer models. We show that the phase separation capacity of the androgen receptor is driven by aromatic residues and short unstable helices in its intrinsically disordered activation domain. Based on this knowledge, we developed tool compounds that covalently attach aromatic moieties to cysteines in the receptors’ activation domain. The compounds enhanced partitioning of the receptor into condensates, facilitated degradation of the receptor, inhibited androgen receptor-dependent transcriptional programs, and had antitumorigenic effect in models of prostate cancer and castration-resistant prostate cancer in vitro and in vivo. These results establish a generalizable framework to target the phase- separation capacity of intrinsically disordered regions in oncogenic transcription factors and other disease-associated proteins with therapeutic intent

Conformational conversion during controlled oligomerization into nonamylogenic protein nanoparticles

Protein materials are rapidly gaining interest in materials sciences and nanomedicine because of their intrinsic biocompatibility and full biodegradability. The controlled construction of supramolecular entities relies on the controlled oligomerization of individual polypeptides, achievable through different strategies. Because of the potential toxicity of amyloids, those based on alternative molecular organizations are particularly appealing, but the structural bases on nonamylogenic oligomerization remain poorly studied. We have applied spectrofluorimetry and spectropolarimetry to identify the conformational conversion during the oligomerization of His-tagged cationic stretches into regular nanoparticles ranging around 11 nm, useful for tumor-targeted drug delivery. We demonstrate that the novel conformation acquired by the proteins, as building blocks of these supramolecular assemblies, shows different extents of compactness and results in a beta structure enrichment that enhances their structural stability. The conformational profiling presented here offers clear clues for understanding and tailoring the process of nanoparticle formation through the use of cationic and histidine rich stretches in the context of protein materials usable in advanced nanomedical strategies.Fil: Sanchez, Julieta Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universitat Autònoma de Barcelona; España. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; ArgentinaFil: Sánchez García, Laura. Universitat Autònoma de Barcelona; España. CIBER de Bioingeniería, Biomateriales y Nanomedicina, Barcelona; EspañaFil: Pesarrodona, Mireia. Universitat Autònoma de Barcelona; España. CIBER de Bioingeniería, Biomateriales y Nanomedicina, Barcelona; EspañaFil: Serna, Naroa. Universitat Autònoma de Barcelona; España. CIBER de Bioingeniería, Biomateriales y Nanomedicina, Barcelona; EspañaFil: Sánchez Chardi, Alejandro. Universitat Autònoma de Barcelona; EspañaFil: Unzueta, Ugutz. Universitat Autònoma de Barcelona; España. CIBER de Bioingeniería, Biomateriales y Nanomedicina, Barcelona; EspañaFil: Mangues, Ramón. CIBER de Bioingeniería, Biomateriales y Nanomedicina, Barcelona; EspañaFil: Vazquez, Esther. Universitat Autònoma de Barcelona; España. CIBER de Bioingeniería, Biomateriales y Nanomedicina, Barcelona; EspañaFil: Villaverde Corrales, Antonio. Universitat Autònoma de Barcelona; España. CIBER de Bioingeniería, Biomateriales y Nanomedicina, Barcelona; Españ

Conformational conversion during controlled oligomerization into nonamylogenic protein nanoparticles

Altres ajuts: protein production and DLS have been partially performed by the ICTS "NANBIOSIS", more specifically by the Protein Production Platform of CIBER-BBN/IBB (http://www. nanbiosis.es/unit/u1-protein-production-platform-ppp/) and the Biomaterial Processing and Nanostructuring Unit (http://www.nanbiosis.es/portfolio/u6-biomaterial-processing-and-nanostructuring-unit/). J.M.S. is a Career Investigator from CONICET (Government of Argentina), N.S. was supported by a predoctoral fellowship from the Government of Navarra, and U.U. received a Sara Borrell postdoctoral fellowship from AGAUR. A.V. received an ICREA ACADEMIA award. This study has been funded by CIBER-BBN (project VENOM4-CANCER) granted to A.V.Protein materials are rapidly gaining interest in materials sciences and nanomedicine because of their intrinsic biocompatibility and full biodegradability. The controlled construction of supramolecular entities relies on the controlled oligomerization of individual polypeptides, achievable through different strategies. Because of the potential toxicity of amyloids, those based on alternative molecular organizations are particularly appealing, but the structural bases on nonamylogenic oligomerization remain poorly studied. We have applied spectrofluorimetry and spectropolarimetry to identify the conformational conversion during the oligomerization of His-tagged cationic stretches into regular nanoparticles ranging around 11 nm, useful for tumor-targeted drug delivery. We demonstrate that the novel conformation acquired by the proteins, as building blocks of these supramolecular assemblies, shows different extents of compactness and results in a beta structure enrichment that enhances their structural stability. The conformational profiling presented here offers clear clues for understanding and tailoring the process of nanoparticle formation through the use of cationic and histidine rich stretches in the context of protein materials usable in advanced nanomedical strategies