Protein-small Molecule Research Articles

524P A phase I, open-label, dose-escalation trial of BI 1701963 in patients (pts) with KRAS mutated solid tumours: A snapshot analysis

BI 1701963 is a small-molecule protein–protein interaction inhibitor, which prevents KRAS activation by binding to the catalytic site of SOS1. This multicentre phase I first-in-human trial is evaluating BI 1701963 as monotherapy (mono tx) or in combination with trametinib in adults with KRAS-mutated solid tumours. Here, we report preliminary results from the mono tx arm, which aims to determine the maximum tolerated dose (MTD) and/or recommended phase II dose of BI 1701963, based on dose-limiting toxicities (DLTs).

Protein–Small Molecule Interactions in Native Mass Spectrometry

Small molecule drug discovery has been propelled by the continual development of novel scientific methodologies to occasion therapeutic advances. Although established biophysical methods can be used to obtain information regarding the molecular mechanisms underlying drug action, these approaches are often inefficient, low throughput, and ineffective in the analysis of heterogeneous systems including dynamic oligomeric assemblies and proteins that have undergone extensive post-translational modification. Native mass spectrometry can be used to probe protein-small molecule interactions with unprecedented speed and sensitivity, providing unique insights into polydisperse biomolecular systems that are commonly encountered during the drug discovery process. In this review, we describe potential and proven applications of native MS in the study of interactions between small, drug-like molecules and proteins, including large multiprotein complexes and membrane proteins. Approaches to quantify the thermodynamic and kinetic properties of ligand binding are discussed, alongside a summary of gas-phase ion activation techniques that have been used to interrogate the structure of protein-small molecule complexes. We additionally highlight some of the key areas in modern drug design for which native mass spectrometry has elicited significant advances. Future developments and applications of native mass spectrometry in drug discovery workflows are identified, including potential pathways toward studying protein-small molecule interactions on a whole-proteome scale.

Advances in Engineered Polymer Nanoparticle Tracking Platforms towards Cancer Immunotherapy-Current Status and Future Perspectives.

Engineering polymeric nanoparticles for their shape, size, surface chemistry, and functionalization using various targeting molecules has shown improved biomedical applications for nanoparticles. Polymeric nanoparticles have created tremendous therapeutic platforms, particularly applications related to chemo- and immunotherapies in cancer. Recently advancements in immunotherapies have broadened this field in immunology and biomedical engineering, where “immunoengineering” creates solutions to target translational science. In this regard, the nanoengineering field has offered the various techniques necessary to manufacture and assemble multifunctional polymeric nanomaterial systems. These include nanoparticles functionalized using antibodies, small molecule ligands, targeted peptides, proteins, and other novel agents that trigger and encourage biological systems to accept the engineered materials as immune enhancers or as vaccines to elevate therapeutic functions. Strategies to engineer polymeric nanoparticles with therapeutic and targeting molecules can provide solutions for developing immune vaccines via maintaining the receptor storage in T- and B cells. Furthermore, cancer immunotherapy using polymeric nanomaterials can serve as a gold standard approach for treating primary and metastasized tumors. The current status of the limited availability of immuno-therapeutic drugs highlights the importance of polymeric nanomaterial platforms to improve the outcomes via delivering anticancer agents at localized sites, thereby enhancing the host immune response in cancer therapy. This review mainly focuses on the potential scientific enhancements and recent developments in cancer immunotherapies by explicitly discussing the role of polymeric nanocarriers as nano-vaccines. We also briefly discuss the role of multifunctional nanomaterials for their therapeutic impacts on translational clinical applications.

Microscale thermophoresis for studying protein-small molecule affinity: Application to hyaluronidase

Studying the binding affinity between large biomolecules (i.e. enzyme) and small molecules (molecular weight less than 1000 Da) is an extremely challenging task when using conventional techniques because of the difficulty associated with the detection of the minuscule variations caused by the binding. Microscale thermophoresis (MST) is a sensitive novel emerging biophysical tool allowing a rapid and accurate determination of the dissociation constant (Kd) of enzyme - small molecule system. For the first time, a systematic MST study was carried out to evaluate Kd between hyaluronidase (Hyal), a model enzyme, and two small flavonoid compounds known as potent Hyal's inhibitors, epigallocatechin gallate (EGCG) and apigenin-7-glucoside. In the optimized conditions, hyaluronidase fluorescent labelling (Hyal*) was achieved by direct labelling with RED-NHS NT647 dye using a phosphate ammonia buffer (20 mM NaH2PO4 + 50 mM NH3) + 77 mM NaCl at pH 6.6. This latter ensured a high labelling efficiency while preventing Hyal* precipitation (pI of hyaluronidase is equal to 8.6). Pre-capillary CE/UV enzymatic assays of Hyal* demonstrated the preservation of its enzymatic activity even though the tested concentration was very low (50-fold times lower than control enzymatic assay). Fluorescence signal intensity and shape as well as MST traces have shown that phosphate ammonia buffer required the addition of surfactant, Tween-20 at 0.05%, to efficiently improve the protein solubility and limited aggregate formation using standard uncoated capillaries. MST results revealed the absence of affinity between Hyal* and the tested flavonoids regardless of the incubation time and the tested inhibitory concentration range unless the hyaluronidase substrate, hyaluronic acid (HA) is added to the preparation. Kd values were estimated to be 163 µM for Hyal*/HA/EGCG and 157 µM for Hyal*/HA/apigenin-7-glucoside. The repeatability of the assays was excellent as confirmed by the very low standard deviation on Kd values.

Template Instrumentation for "Accurate Constant via Transient Incomplete Separation".

Accurate Constant via Transient Incomplete Separation (ACTIS) is a new method for finding the equilibrium dissociation constant Kd of a protein-small molecule complex based on transient incomplete separation of the complex from the unbound small molecule in a capillary. This separation is caused by differential transverse diffusion of the complex and the small molecule in a pressure-driven flow. The advection-diffusion processes underlying ACTIS can be described by a system of partial differential equations allowing for a virtual ACTIS instrument to be built and ACTIS to be studied in silico. The previous in silico studies show that large variations in the fluidic system geometry do not affect the accuracy of Kd determination, thus, proving that ACTIS is conceptually accurate. The conceptual accuracy does not preclude, however, instrumental inaccuracy caused by run-to-run signal drifts. Here we report on assembling a physical ACTIS instrument with a fluidic system that mimics the virtual one and proving the absence of signal drifts. Furthermore, we confirmed method ruggedness by assembling a second ACTIS instrument and comparing the results of experiments performed with both instruments in parallel. Despite some unintentional differences between the instruments (caused by tolerances in sizes, positions, etc.) and noticeable differences in their respective separagrams, we found that the Kd values determined for identical samples with these instruments were equal. Conclusively, the fluidic system presented here can serve as a template for reliable ACTIS instrumentation.

Effect of inoculation of starter on physicochemical properties and texture characteristics of fermented beef jerky

In this paper, a selected starter culture of Staphylococcus xylosus, Pediococcus acidilactici, and Debaryomyces hansenii was used to produce fermented beef jerky. The influence of inoculating compound starter on the basic nutrition composition, physical and chemical properties, and texture characteristics of jerkies were investigated. The study results demonstrated that the addition of compound starter quickly reduced the beef pH (from 6.46 to 4.42) and increased the total free amino acids and free fatty acids contents. The nitrite residues and histamine content in the fermentation group were significantly lower than that in the control group. The gel electrophoresis pattern showed that part of the macromolecular protein in inoculated samples was gradually degraded into small molecule protein, and some bands disappeared. It can be seen that inoculation of starter can improve the quality, safety, and texture characteristics of fermented beef jerky. Practical applications Microbial fermentation is a unique and long-standing way of meat processing and preservation. This study found that the inoculation of compound starters can increase the nutritional value and improve texture, flavor, safety of beef jerky. These results suggest that Staphylococcus xylosus, Pediococcus acidilactici, and Debaryomyces hansenii could be considered as mixed bacterial starters in fermented beef jerky to improve product quality and safety, which provides a theoretical basis for the strain screening and processing of fermented beef products.

Engineered protein-small molecule conjugates empower selective enzyme inhibition

Potent, specific ligands drive precision medicine and fundamental biology. Proteins, peptides, and small molecules constitute effective ligand classes. Yet greater molecular diversity would aid the pursuit of ligands to elicit precise biological activity against challenging targets. We demonstrate a platform to discover protein-small molecule (PriSM) hybrids to combine unique pharmacophore activities and shapes with constrained, efficiently engineerable proteins. In this platform, a fibronectin protein library is displayed on yeast with a single cysteine coupled to acetazolamide via a maleimide-poly(ethylene glycol) linker. Magnetic and flow cytometric sorts enrich specific binders to carbonic anhydrase isoforms. Isolated PriSMs exhibit potent, specific inhibition of carbonic anhydrase isoforms with efficacy superior to that of acetazolamide or protein alone, including an 80-fold specificity increase and 9-fold potency gain. PriSMs are engineered with multiple linker lengths, protein conjugation sites, and sequences against two different isoforms, which reveal platform flexibility and impacts of molecular designs. PriSMs advance the molecular diversity of efficiently engineerable ligands.

Impact of Selected Small-Molecule Kinase Inhibitors on Lipid Membranes.

Small-molecule protein kinase inhibitors are used for the treatment of various diseases. Although their effect(s) on the respective kinase are generally quite well understood, surprisingly, their interaction with membranes is only barely investigated; even though these drugs necessarily come into contact with the plasma and intracellular membranes. Using biophysical methods such as NMR, ESR, and fluorescence spectroscopy in combination with lipid vesicles, we studied the membrane interaction of the kinase inhibitors sunitinib, erlotinib, idelalisib, and lenvatinib; these drugs are characterized by medium log p values, a parameter reflecting the overall hydrophobicity of the molecules, which is one important parameter to predict the interaction with lipid membranes. While all four molecules tend to embed in a similar region of the lipid membrane, their presence has different impacts on membrane structure and dynamics. Most notably, sunitinib, exhibiting the lowest log p value of the four inhibitors, effectively influences membrane integrity, while the others do not. This shows that the estimation of the effect of drug molecules on lipid membranes can be rather complex. In this context, experimental studies on lipid membranes are necessary to (i) identify drugs that may disturb membranes and (ii) characterize drug–membrane interactions on a molecular level. Such knowledge is important for understanding the efficacy and potential side effects of respective drugs.

Protein-Peptide Binding Site Detection Using 3D Convolutional Neural Networks.

Peptides and peptide-based molecules represent a promising therapeutic modality targeting intracellular protein-protein interactions, potentially combining the beneficial properties of biologics and small-molecule drugs. Protein-peptide complexes occupy a unique niche of interaction interfaces with respect to protein-protein and protein-small molecule complexes. Protein-peptide binding site identification resembles image object detection, a field that had been revolutionalized with computer vision techniques. We present a new protein-peptide binding site detection method called BiteNetPp by harnessing the power of 3D convolutional neural network. Our method employs a tensor-based representation of spatial protein structures, which is fed to 3D convolutional neural network, resulting in probability scores and coordinates of the binding "hot spots" in the input structures. We used the domain adaptation technique to fine-tune model trained on protein-small molecule complexes using a manually curated set of protein-peptide structures. BiteNetPp consistently outperforms existing state-of-the-art methods in the independent test benchmark. It takes less than a second to analyze a single-protein structure, making BiteNetPp suitable for the large-scale analysis of protein-peptide binding sites.

Protein Phosphatase 1 Regulates Human Cytomegalovirus Protein Translation by Restraining AMPK Signaling

Human cytomegalovirus (HCMV) carries the human protein phosphatase 1 (PP1) and other human proteins important for protein translation in its tegument layer for a rapid supply upon infection. However, the biological relevance behind PP1 incorporation and its role during infection is unclear. Additionally, PP1 is a difficult molecular target due to its promiscuity and similarities between the catalytic domain of multiple phosphatases. In this study, we circumvented these shortcomings by using 1E7-03, a small molecule protein–protein interaction inhibitor, as a molecular tool of noncatalytic PP1 inhibition. 1E7-03 treatment of human fibroblasts severely impaired HCMV replication and viral protein translation. More specifically, PP1 inhibition led to the deregulation of metabolic signaling pathways starting at very early time points post-infection. This effect was at least partly mediated by the prevention of AMP-activated protein kinase dephosphorylation, leading to elongation factor 2 hyperphosphorylation and reduced translation rates. These findings reveal an important mechanism of PP1 for lytic HCMV infection.

Abstract 2890: Mitotic inhibitors suppresses tumor formation and metastasis of triple negative breast cancer (TNBC)

Abstract Cell division cycle 20 (Cdc20) is the main activator of the anaphase-promoting complex (APC/C) and is essential in mitosis regulation. Recently, several papers reported that Cdc20 is highly conserved in various human cancers such as pancreatic, prostate, and lung cancers yet its relation to breast cancer is unclear. Here, we found that Cdc20 gene was highly expressed in three different TNBC cell lines among the mitosis regulator genes through the National Center for Biotechnology Information (NCBI). High Cdc20 level is positively correlated with metastasis-free patient survival and relapse-free patient survival in breast cancer patient database. We also checked that Cdc20 is highly conserved in all TNBC cell lines compared to other subtypes of breast cancer cell lines and the knockdown of this gene decreased cell growth and invasion in TNBC. Furthermore, we investigated that the mitosis-related small molecule protein kinase inhibitors such as VX-680, ZM447439, BI2536, and apcin also inhibit tumor cell proliferation, migration, and invasion. In conclusion, Cdc20 has an important role in TNBC cell growth and metastasis. Cdc20 might serve as a potential marker of disease progression and as a principal potential therapeutic target. Also, mitosis-related protein kinase inhibitors are additional potential therapy to suppress cancer cell progression and invasion in TNBC effectively. Citation Format: Chaeun Song, SeungBaek Lee. Mitotic inhibitors suppresses tumor formation and metastasis of triple negative breast cancer (TNBC) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2890.

Advancements in Protein based Nano Particulate system for treatment of Pulmonary Infections- A Review

In addition to the so-called small molecule drugs, proteins and peptides are of increasing interest forpharmacotherapy, due to several advantageous properties. In general, those compounds are administered parenterally. However, non-invasive routes of administration represent a great part of research. Amongst others is the pulmonary application of proteins and peptides for local delivery in the case of pulmonary diseases, such as idiopathic pulmonary fibrosis, where the alveolar epithelium is affected. To ensure an intracellular delivery, nano particles in a size range of 150 nm will be prepared via charge-mediated coacervation, characterized for their physicochemical properties and loaded with several model-proteins. The material used for nano particle preparation was chosen to be positively and negatively charged starch derivatives, which were synthesized from potato starch. Although nano particles in that size range are known to show an increased cell uptake, they do not show a high deposition in the deep lung. Thus, an advanced carrier system consisting of a fast dissolving micro particle matrix with embedded starch nano particles will be developed and characterized. Due to its aerodynamic properties, that carrier system must be able to deposit a high fraction of the applied dose in the deep lung (~50%), while at the same time demonstrating (in in vitro models) the ability to facilitate uptake of starch nano particles into cells of the alveolar epithelium after fast dissolution of the micro particle matrix.

Absorption Mode Fourier Transform Ion Mobility Mass Spectrometry Multiplexing Combined with Half-Window Apodization Windows Improves Resolution and Shortens Acquisition Times.

Fourier transform multiplexing enables the coupling of drift tube ion mobility to a wide array of mass spectrometers with improved ion utilization and duty cycles compared to dual-gate signal averaging methods. Traditionally, the data generated by this method is presented in the magnitude mode, but significant improvements in resolution and the signal-to-noise ratio (SNR) are expected if the data can be phase corrected and presented in the absorption mode. A method to simply and reliably determine and correct phase shifts in Fourier transform ion mobility mass spectrometry data using information readily available to any user is presented and evaluated for both small molecule and intact protein analyses with no modification to instrument hardware or experimental procedures. Additionally, the effects of apodization and zero padding are evaluated for both processing methods, and a strategy to use these techniques to reduce acquisition times is presented and evaluated. Resolution is improved by an average factor of 1.6, the SNR is improved by an average factor of 1.2, and acquisition times are reduced by up to 80% through the application of absorption mode processing combined with apodization and zero padding.

A covariation analysis reveals elements of selectivity in quorum sensing systems.

Many bacteria communicate with kin and coordinate group behaviors through a form of cell-cell signaling called acyl-homoserine lactone (AHL) quorum sensing (QS). In these systems, a signal synthase produces an AHL to which its paired receptor selectively responds. Selectivity is fundamental to cell signaling. Despite its importance, it has been challenging to determine how this selectivity is achieved and how AHL QS systems evolve and diversify. We hypothesized that we could use covariation within the protein sequences of AHL synthases and receptors to identify selectivity residues. We began by identifying about 6000 unique synthase-receptor pairs. We then used the protein sequences of these pairs to identify covariation patterns and mapped the patterns onto the LasI/R system from Pseudomonas aeruginosa PAO1. The covarying residues in both proteins cluster around the ligand-binding sites. We demonstrate that these residues are involved in system selectivity toward the cognate signal and go on to engineer the Las system to both produce and respond to an alternate AHL signal. We have thus demonstrated that covariation methods provide a powerful approach for investigating selectivity in protein-small molecule interactions and have deepened our understanding of how communication systems evolve and diversify.

'All That Glitters Is Not Gold': High-Resolution Crystal Structures of Ligand-Protein Complexes Need Not Always Represent Confident Binding Poses.

Our understanding of the structure–function relationships of biomolecules and thereby applying it to drug discovery programs are substantially dependent on the availability of the structural information of ligand–protein complexes. However, the correct interpretation of the electron density of a small molecule bound to a crystal structure of a macromolecule is not trivial. Our analysis involving quality assessment of ~0.28 million small molecule–protein binding site pairs derived from crystal structures corresponding to ~66,000 PDB entries indicates that the majority (65%) of the pairs might need little (54%) or no (11%) attention. Out of the remaining 35% of pairs that need attention, 11% of the pairs (including structures with high/moderate resolution) pose serious concerns. Unfortunately, most users of crystal structures lack the training to evaluate the quality of a crystal structure against its experimental data and, in general, rely on the resolution as a ‘gold standard’ quality metric. Our work aims to sensitize the non-crystallographers that resolution, which is a global quality metric, need not be an accurate indicator of local structural quality. In this article, we demonstrate the use of several freely available tools that quantify local structural quality and are easy to use from a non-crystallographer’s perspective. We further propose a few solutions for consideration by the scientific community to promote quality research in structural biology and applied areas.

Nanoemulsions for drug delivery

Emulsions are liquid–liquid dispersions with one liquid phase dispersed in the other liquid phase as small droplets. Nanoemulsions are nano-sized emulsions with sizes ranging from tens to hundreds of nanometers, and have great potential applications in pharmaceutics, foods and cosmetics due to their attractive properties, such as small sizes, high surface area per unit volume, improved dispersion of active hydrophobic components and enhanced absorption. The article provides an overview of nanoemulsions for drug delivery, starting with an introduction of emulsion types, nanoemulsion preparation and nanoemulsion stability. Surfactants play critical roles in producing and stabilizing nanoemulsions. Different types of surfactants are summarized including small molecule surfactants, particle surfactants, phospholipids, peptide and protein surfactants. Then the applications of nanoemulsions as nanomedicine in drug delivery are presented. Finally, clinical applications of nanoemulsions are discussed.

An Overview of the Application of Poly(lactic-co-glycolic) Acid (PLGA)-Based Scaffold for Drug Delivery in Cartilage Tissue Engineering

Poly(lactic-co-glycolic) acid (PLGA) has attracted a considerable amount of interest for biomedical application due to its biocompatibility, tailored biodegradation rate (depending on the molecular weight and copolymer ratio), approval for clinical use in humans by the U.S. Food and Drug Administration (FDA), the potential to change surface properties to create better interaction with biological materials and being suitable for export to countries and cultures where planting products with animals is unusable. For commercial use and research, PLGA has been widely studied to control small molecule drugs, proteins, and other macromolecules. This study aims to review the studies that used PLGA scaffolding and its composites as a scaffold and drug delivery in cartilage tissue engineering. It is concluded from the results that the PLGA scaffold as a synthetic scaffold, when combined with natural scaffolds or hybrids, strengthens its biological properties and performs its function better.

The role of dermatopontin in cell adhesion in bovine placenta during early-mid pregnancy and parturition – Pilot study

Dermatopontin (DPT) is a small protein molecule thought to have a role in the formation of the extracellular architecture and adhesion. The aim of the study was to confirm the presence of DPT and to examine its role in placental cell adhesion during pregnancy, at parturition and postpartum in cows.Placental tissue samples were obtained at abattoir from healthy pregnant cows (n = 6) while parturient samples were collected during caesarian section and retrospectively divided into released up to 6 h (R; n = 5) and not released up to 6 h (NR; n = 4) foetal membranes. Maternal epithelial cells were isolated from pregnant samples and were used for the examination of the influence of DPT (5, 50 and 100 ng/mL) on cell adhesion. Parturient samples were manually divided into maternal and foetal part and individually homogenized for Western blotting and ELISA analysis.Western blotting confirmed the presence of DPT in examined tissues. ELISA test showed significant (p < 0.05) decrease in DPT concentration within examined pregnancy period with higher concentrations in maternal part (p < 0.05). Moreover, at parturition DPT concentration further decreased in maternal (p < 0.05) but increased (p < 0.05) in fetal part. The examination of not released samples showed opposite relationship in comparison to parturient samples – the increase in maternal (p < 0.05) and the decrease in fetal (p < 0.05) part of placenta. DPT facilitated the adhesion of epithelial cells in examined periods of pregnancy in increasing manner with pregnancy course.The presence of DPT in bovine placenta during pregnancy and parturition was confirmed. This protein may influence cell adhesion during attachment and detachment of placenta. Further studies on mechanisms of action of DPT in bovine placenta are necessary.

Roles of exosomes in cancer chemotherapy resistance, progression, metastasis and immunity, and their clinical applications (Review)

Exosomes are a type of vesicle that are secreted by cells, with a diameter of 40-100 nm, and that appear as a cystic shape under an electron microscope. Exosome cargo includes a variety of biologically active substances such as non-coding RNA, lipids and small molecule proteins. Exosomes can be taken up by neighboring cells upon secretion or by distant cells within the circulatory system, affecting gene expression of the recipient cells. The present review discusses the formation and secretion of exosomes, and how they can remodel the tumor microenvironment, enhancing cancer cell chemotherapy resistance and tumor progression. Exosome-mediated induction of tumor metastasis is also highlighted. More importantly, the review discusses the manner in which exosomes can change the metabolism of cancer cells and the immune system, which may help to devise novel therapeutic approaches for cancer treatment. With the development of nanotechnology, exosomes can also be used as biomarkers and for the delivery of chemical drugs, serving as a tool to diagnose and treat cancer.

The small-molecule protein ligand interface stabiliser E7820 induces differential cell line specific responses of integrin \u03b12 expression

BackgroundThe mechanism of small-molecule stabilised protein-protein interactions is of growing interest in the pharmacological discovery process. A plethora of different substances including the aromatic sulphonamide E7820 have been identified to act by such a mechanism. The process of E7820 induced CAPERα degradation and the resultant transcriptional down regulation of integrin α2 expression has previously been described for a variety of different cell lines and been made responsible for E7820’s antiangiogenic activity. Currently the application of E7820 in the treatment of various malignancies including pancreas carcinoma and breast cancer is being investigated in pre-clinical and clinical trials. It has been shown, that integrin α2 deficiency has beneficial effects on bone homeostasis in mice. To transfer E7820 treatment to bone-related pathologies, as non-healing fractures, osteoporosis and bone cancer might therefore be beneficial. However, at present no data is available on the effect of E7820 on osseous cells or skeletal malignancies.MethodsPre-osteoblastic (MC3T3 and Saos-2) cells and endothelial (eEnd2 cells and HUVECs) cells, each of human and murine origin respectively, were investigated. Vitality assay with different concentrations of E7820 were performed. All consecutive experiments were done at a final concentration of 50 ng/ml E7820. The expression and production of integrin α2 and CAPERα were investigated by quantitative real-time PCR and western blotting. Expression of CAPERα splice forms was differentiated by semi-quantitiative reverse transcriptase PCR.ResultsHere we present the first data showing that E7820 can increase integrin α2 expression in the pre-osteoblast MC3T3 cell line whilst also reproducing canonical E7820 activity in HUVECs. We show that the aberrant activity of E7820 in MC3T3 cells is likely due to differential activity of CAPERα at the integrin α2 promoter, rather than due to differential CAPERα degradation or differential expression of CAPERα spliceforms.ConclusionThe results presented here indicate that E7820 may not be suitable to treat certain malignancies of musculoskeletal origin, due to the increase in integrin α2 expression it may induce. Further investigation of the differential functioning of CAPERα and the integrin α2 promoter in cells of various origin would however be necessary to more clearly differentiate between cell lines that will positively respond to E7820 from those that will not.