High-Throughput Screening Assays to Discover Small-Molecule Inhibitors of Protein Interactions

Colas, Pierre

doi:10.2174/157016308785739875

Cited by 32 publications

(40 citation statements)

References 57 publications

(71 reference statements)

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“…The next step involves feasibility and bioactivity testing and should be carried out in vitro and in vivo. In vitro assays using cell culture preparations are used to characterise the effects of materials on isolated cell function and for screening large numbers of compounds for biological activity, toxicity and immunogenicity (193)(194). However, due to their nature using isolated cells, in vitro models are unavoidably limited in their capacity to reflect complex in vivo environments that the TEC will be exposed to and are therefore inadequate to predict in vivo or clinical performances.…”

Section: Rationale For Translating Bone Tissue Engineering Strategiesmentioning

confidence: 99%

Bone Regeneration Based on Tissue Engineering Conceptions — A 21st Century Perspective

et al. 2013

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The role of Bone Tissue Engineering in the field of Regenerative Medicine has been the topic of substantial research over the past two decades. Technological advances have improved orthopaedic implants and surgical techniques for bone reconstruction. However, improvements in surgical techniques to reconstruct bone have been limited by the paucity of autologous materials available and donor site morbidity. Recent advances in the development of biomaterials have provided attractive alternatives to bone grafting expanding the surgical options for restoring the form and function of injured bone. Specifically, novel bioactive (second generation) biomaterials have been developed that are characterised by controlled action and reaction to the host tissue environment, whilst exhibiting controlled chemical breakdown and resorption with an ultimate replacement by regenerating tissue. Future generations of biomaterials (third generation) are designed to be not only osteoconductive but also osteoinductive, i.e. to stimulate regeneration of host tissues by combining tissue engineering and in situ tissue regeneration methods with a focus on novel applications. These techniques will lead to novel possibilities for tissue regeneration and repair. At present, tissue engineered constructs that may find future use as bone grafts for complex skeletal defects, whether from post-traumatic, degenerative, neoplastic or congenital/developmental "origin" require osseous reconstruction to ensure structural and functional integrity. Engineering functional bone using combinations of cells, scaffolds and bioactive factors is a promising strategy and a particular feature for future development in the area of hybrid materials which are able to exhibit suitable biomimetic and mechanical properties. This review will discuss the state of the art in this field and what we can expect from future generations of bone regeneration concepts.

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Section: Rationale For Translating Bone Tissue Engineering Strategiesmentioning

confidence: 99%

Bone Regeneration Based on Tissue Engineering Conceptions — A 21st Century Perspective

et al. 2013

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“…PPIs have enormous potential as targets for drug discovery, because they are obligatory for all cellular functions. [52][53][54][55][56] Structural studies indicate that protein-binding interfaces contain discrete ''hot spots'' where a relatively small number of amino acids at the PPI interface contribute the majority of the binding energy, and that contact surfaces contain cavities, pockets, and grooves for small molecules to bind to and disrupt PPIs. 52,53,55,56 TIF2 is an AR coactivator implicated in the development and progression of CRPC.…”

Section: 46mentioning

confidence: 99%

“…[52][53][54][55][56] Structural studies indicate that protein-binding interfaces contain discrete ''hot spots'' where a relatively small number of amino acids at the PPI interface contribute the majority of the binding energy, and that contact surfaces contain cavities, pockets, and grooves for small molecules to bind to and disrupt PPIs. 52,53,55,56 TIF2 is an AR coactivator implicated in the development and progression of CRPC. 5,21,28,29 Agonist-induced conformational changes to the AR-LBD create the activation function 2 (AF2) surface that binds to the LXXLL motifs of SRC/p160 coactivators.…”

Section: 46mentioning

confidence: 99%

Reconfiguring the AR-TIF2 Protein–Protein Interaction HCS Assay in Prostate Cancer Cells and Characterizing the Hits from a LOPAC Screen

Fancher

Hua

Camarco

et al. 2016

ASSAY and Drug Development Technologies

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The continued activation of androgen receptor (AR) transcription and elevated expression of AR and transcriptional intermediary factor 2 (TIF2) coactivator observed in prostate cancer (CaP) recurrence and the development of castration-resistant CaP (CRPC) support a screening strategy for small-molecule inhibitors of AR-TIF2 protein-protein interactions (PPIs) to find new drug candidates. Small molecules can elicit tissue selective effects, because the cells of distinct tissues express different levels and cohorts of coregulatory proteins. We reconfigured the AR-TIF2 PPI biosensor (PPIB) assay in the PC-3 CaP cell line to determine whether AR modulators and hits from an AR-TIF2 PPIB screen conducted in U-2 OS cells would behave differently in the CaP cell background. Although we did not observe any significant differences in the compound responses between the assay performed in osteosarcoma and CaP cells, the U-2 OS AR-TIF2 PPIB assay would be more amenable to screening, because both the virus and cell culture demands are lower. We implemented a testing paradigm of counter-screens and secondary hit characterization assays that allowed us to identify and deprioritize hits that inhibited/disrupted AR-TIF2 PPIs and AR transcriptional activation (AR-TA) through antagonism of AR ligand binding or by non-specifically blocking nuclear receptor trafficking. Since AR-TIF2 PPI inhibitor/disruptor molecules act distally to AR ligand binding, they have the potential to modulate AR-TA in a cell-specific manner that is distinct from existing anti-androgen drugs, and to overcome the development of resistance to AR antagonism. We anticipate that the application of this testing paradigm to characterize the hits from an AR-TIF2 PPI high-content screening campaign will enable us to prioritize the AR-TIF2 PPI inhibitor/disruptor leads that have potential to be developed into novel therapeutics for CaP and CRPC.

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“…More recently also cellular assays based on the 2-hybrid concept became available that are 11 compatible with the scale required for HTS [60,61]. In these assays, two interacting proteins of interest are fused to complementary fragments of a reporter protein (e.g.…”

Section: Filling the Pipeline Of Ppi Modulatory Drugsmentioning

confidence: 99%

Protein-protein Interactions: Network Analysis and Applications in Drug Discovery

Bultinck¹,

Lievens²,

Tavernier

2012

CPD

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Physical interactions among proteins constitute the backbone of cellular function, making them an attractive source of therapeutic targets. Although the challenges associated with targeting proteinprotein interactions (PPIs) -in particular with small molecules -are considerable, a growing number of functional PPI modulators is being reported and clinically evaluated. An essential starting point for PPI inhibitor screening or design projects is the generation of a detailed map of the human interactome and the interactions between human and pathogen proteins. Different routes to produce these biological networks are being combined, including literature curation and computational methods. Experimental approaches to map PPIs mainly rely on the yeast two-hybrid (Y2H) technology, which have recently shown to produce reliable protein networks. However, other genetic and biochemical methods will be essential to increase both coverage and resolution of current protein networks in order to increase their utility towards the identification of novel disease-related proteins and PPIs, and their potential use as therapeutic targets.

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High-Throughput Screening Assays to Discover Small-Molecule Inhibitors of Protein Interactions

Cited by 32 publications

References 57 publications

Bone Regeneration Based on Tissue Engineering Conceptions — A 21st Century Perspective

Bone Regeneration Based on Tissue Engineering Conceptions — A 21st Century Perspective

Reconfiguring the AR-TIF2 Protein–Protein Interaction HCS Assay in Prostate Cancer Cells and Characterizing the Hits from a LOPAC Screen

Protein-protein Interactions: Network Analysis and Applications in Drug Discovery

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