Both NGR and RGD conjugated forms of ketoprofen showed higher cytotoxic activity against OVCAR3 and HT-1-80 respectively.It is well known that Arginine-Glycine-Aspartic acid (RGD) and Asparagine-Glycine-Arginine (NGR) peptides preferentially bind to integrin receptors and aminopeptidase N respectively and these two receptors play important roles in angiogenesis. Therefore ketoprofen as a non-selective cox Inhibitor was conjugated with linear RGD and NGR to take advantage of targeting capability of these two motifs and delivering ketoprofen to these cancer cells with overexpression of integrin and aminopeptidase N. In order to investigate the impact of possible steric hindrance due to the attachment of the drug to the peptide, a linear six carbon (hexanoic acid) linker was also used as a spacer. Cytotoxic effect of the synthesized compounds was evaluated against a group of cancer cell lines, including MCF-7, A2780 (α Cancer is unusual cell growth and division which can spread to different parts of body. Chemotherapy is a main approach for cancer treatment and limited by low selectivity of chemotherapy drugs for normal tissues, especially tissues with rapidly cells division and growth.v integrin is highly overexpressed on activated endothelial of cancer cells and tumor cells, but it is not expressed in resting endothelial cells and most normal organ systems, making it a potential target for antiangiogenic strategy in cancer therapy  and NGR (Asparagine-Glycine-Arginine) motifs have been recognized as targeting tools based on phage display studies . RGD is er cells . Since αer cells -19, RGD er cells . NGR pepet al. have demonstrated that ketoprofen inhibits human colon tumor cell N-acetyltransferase (NAT) activity, gene expression and DNA adduct formation for the first time in 2006  which inhibits COX-1 and COX-2 enzymes. Numerous studies demonstrate that NSAIDs have anticancer properties and reduced the risk of cancers like colon and liver  Several  in 2006 . Marjano in 2006 . Based oMaterials and methodsa-9-Fluorenylmethoxycarbonyl (Fmoc)-protected amino acids, Rink amid resin, 1-[Bis(dimethylamino)methylene]-1H- 1,2,3-triazolo pyridinium 3-oxid hexafluorophosphate (HATU) were purchased from GL Biochem Company (Shanghi) Ltd. 1HNMR spectra were recorded on a 500 MHz Bruker spectrometer. ESI-MS spectra were obtained by Agilent 6410 Triple Quad LC/MS. IR spectra were recorded on a PerkinElmer IR spectrophotometer as potassium bromide discs. The purity of compounds was confirmed by thin layer chromatography using Whatman Sil All chemicals and solvents were purchased from Merck . N® HR ODS column .G/UV254 silica gel plates as the stationary phase and with suitable mobile phase with fluorescent indicator, and the spots were visualized under 254 and 366 nm illumination. All products was purified by reverse phase HPLC on a Knaur chromatography system equipped with a Waters Prep Nova-PakGeneral procedure for synthesis RGD conjugated with ketoprofen2 gas for 20 min. The completion of each coupling reaction and fmoc deprotection was tested by Kaiser Test which is the reaction of ninhydrin with amines. Rink amide resin (0.5 g) was placed in a reactor and suspended in DMF under nitrogen atmosphere for 15 min. The fmoc group was removed with 20% piperidine in DMF (3 × 5 mL). After washing with DMF, coupling reaction was conducted. For each amino acid coupling: 2eq AA, 2eq HATU, 4eq N,N-diisopropylethylamine (DIEA), 15 mL DMF was mixed and added to the resin. The mixture was purged with NAfter the last amino acid coupling reaction and removal of fmoc protecting group, the ketoprofen was coupled to the free amino group of the peptide, using the same standard procedure which is used for the coupling of amino acids. Finally resin washed and dried then linear peptide cleaved with trifluoroacetic acid (TFA) cocktail: TFA: Thioanisol: p-cresol: triisopropylsilane (TIS) (84%: 12%: 2%: 2%), during 2 h at room temperature. The reaction mixture was filtered and cold Diethyl Ether (50 mL) was added to the filtrate. The precipitate thus obtained was separated using centrifugation.RGD 3CN in H2O+ 0.1% TFA); 1H NMR  δ: 1.54 , 1.70 , 2.63-2.69 , 3.11 , 3.86 , 4.50-4.53 , 7.15-8.68 ppm ; IR (KBr): υ (cm-1) = 1656, 3330; LC-MS (ESI) m/z (M + H)+ calcd for C12H23N7O5: 346.0, found: 346.1. The title compound (50 mg) was obtained as a white solid with 63% yield at 97% purity ; 1H NMR  δ: 1.50 , 1.99 , 2.5 , 2.58-2.69 , 3.23 , 4.03 , 7.15-8.3 ppm ; IR (KBr): υ (cm-1) = 1656, 3330; LC-MS (ESI) m/z (M + H)+ calcd for C12H23N7O5: 346.0, found: 346.1.The title compound (57 mg) was obtained as a white solid with 64% yield at 93% purity ; 1H NMR  δ: 1.32-1.33, 1.45-1.46 , 1.64-1.82 , 2.5 , 2.96,3.07 , 3.64-3.69 , 3.82-3.85 , 4.15-4.16 , 4.26-4.27 , 4.38-4.43 , 7.05-8.31, 11.8  ppm; IR (KBr): υ (cm-1) 1656, 3330 ; LC-MS (ESI) m/z (M + H)+ calcd for C28H35N7O7: 581, found: 582.2. The title compound (83 mg) was obtained as a white solid with yield 65%, 98% purity 4-CO-RGD3CN in H2O+ 0.1% TFA); 1H NMR  δ: 1.05 , 1.20-1.23 , 1.28 , 1.48 , 1.7 , 2.07 , 2.8 , 3.17 , 3.38 , 3.46-3.52 , 4.16 , 4.55 , 5.88-5.93 , 7.00-7.8 ppm , IR (KBr): υ (cm-1) 1654, 3356 ; LC-MS (ESI) m/z (M + H)+ calcd for C34H47N9O7: 693, found: 694.2.The title compound (103 mg) was obtained as a white solid with yield 65%, 98% purity ; 1H NMR  δ: 1.05 , 1.25 , 1.45 , 2.5 , 2. 6 , 3.47 , 3.6 , 3.9 , 4.16 , 4.43 , 7.05-8.29 , 11.8  ppm; IR (KBr): υ (cm-1) 1656, 3330; LC-MS (ESI) m/z (M + H)+ calcd for C28H36N8O7: 581, found: 582.2. The title compound (73 mg) was obtained as a white solid with yield 57%, 98% purity 4-CO-NGR3CN in H2O+ 0.1% TFA); 1H NMR  δ: 1.05 , 1.19-1.25 , 1.46-1.48 , 1.64 , 2.28 , 2.49 , 2.90 , 2.97 , 3.40 , 3.46-3.51, 4.12 , 4.8  5.88-5.93 , 7.00-8.03 ppm ; IR (KBr): υ (cm-1); IR (KBr): υ (cm-1) 1659, 1647, 3331; LC-MS (ESI) m/z (M + H)+ calcd for C34H47N9O7: 693, found 694.1. The title compound (110 mg) was obtained as a white solid with yield 69%, 97% purity , OVCAR-3 (high αVβ3), SKOV-3 (low CD13), HT-1080 (high CD13) and MCF-7 and fibroblast cell were selected. A2780 and MCF-7 are both cancer cells, but A2780 is αv β3 integrin overexpressing cell line. Three concentrations of products were prepared and were evaluated after 48 h by using MTT method. Therefore cells cultured in culture media (RPMI1640) at 37 °C under 5% CO2 supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin and 100 μg/mL streptomycin. Nearly 104 cells were seeded into 96-well plates and were grown for 24 h in incubator. Compounds and doxorubicin added to wells, cells were incubated at 37 °C for 48 h. Cells without any adding of compound were used as negative control, Doxorubicin was used as positive control and DMSO as the solvent of the test compounds. After 48 h, 10 μL of MTT (5 mg/mL in PBS) was added at 37 °C for 4 h. The medium with MTT was removed and 100 μL DMSO was added to each well, cover the plate from light, shake it for 5 min, then optical densities were read at 570 nm with a reference wavelength of 630 nm as background using a spectrophotometer plate reader .In order to evaluate the cytotoxic activity of the synthesis compounds, five cell lines: A2780 (low αMolecular modeling (docking) studies vβ3 integrin receptor in the presence of the Mn2+ ion and with active sites of APN. The crystal structure of the extracellular domain of the αvβ3 integrin receptor in the presence of the Mn2+ ion (PDB entry code: 1L5G)    was obtae: 2DQM) .vβ3 integrin receptor. The quality of the docked structures was assessed by measuring the intermolecular energy of the ligand-enzyme assembly. Grid map dimensions for 2DQM (71.9186 × 47.8031 × 7.3077) were set surrounding active site. The energy minimized ligands were docked in binding site of APN.All the compounds were built using hyperchem version 8 and subsequently minimized. The protein structure was prepared for docking using AutoDock Tool. Polar hydrogen was added and non-polar hydrogen was merged and finally Kollman united atom charge and atom type parameters were added to 1L5G for RGD products and 2DQM for NGR products. Grid map dimensions for 1L5G (18.6378 × 41.9200 × 43.1259) were set surrounding active site. The energy minimized ligands were docked in binding site of αChemistryKetoprofen were selected as NSAIDs and fmoc peptide synthesis strategy was used to make both the peptide-drug conjugates and peptide-spacer-drug conjugates. Therefore, fmoc-6-amino hexanoic acid was needed to make the peptide-spacer-drug conjugates . The synBiological activityRGD and NGR are promising peptide sequences for targeted drug delivery to tumor cells. They can carry their drug conjugated to their target tumor. Ketoprofen was attached to N-terminus of the tripeptides RGD and NGR through its carboxylic acid group and thus an amide bond is responsible for the conjugation. In order to avoid the possible steric hindrance which is caused by the closeness of ketoprofen to the tripeptide, another from of the conjugate was also synthesized in which ketoprofen was attached to the tripeptide through a linear six carbon linker. Anti-proliferation activities of the synthesized compound were evaluated on a selected group of cancer cell lines. The results are summarized in a. No inhibitory activity was observed on MCF-7 (as a tumor cell without over-expressed RGD- or NGR-binding protein on its surface) or Fibroblasts .vβ3) on their surface. In case of A2780, keto-spacer-RGD showed higher inhibitory activity than keto-RGD (63 vs. 55% inhibition) which indicates possible role of spacer either in better binding to the receptor or in mitigating the steric hindrance for better binding of RGD to the cell surface. Surprisingly, the opposite pattern is observed in case of OVCAR-3 which shows the possible impediment of linker group for the ligand-receptor binding. b. ketoprofen caused no inhibition at 100 µm concentration either on A2780 or on OVCAR-3. However the conjugated forms of ketoprofen with RGD either with or without spacer, showed >50% inhibition on both cell lines. The two cell lines are well recognized for the overexpression of RGD receptors (A2780 with low and OVCAR-3 with high expression of αc. NGR conjugates of ketoprofen caused 3-7 times higher inhibition compared to ketoprofen on HT-1080 and SKOV-3. These are the cancer cell lines with over expression of NGR receptor (SKOV-3 with low and HT-1080 with high expression of CD13) on their surface. Therefore it could be speculated that the NGR companionship with ketoprofen has improved its cytotoxic activity on these cell lines. In both cases, the conjugated form which contains the spacer between ketoprofen and NGR showed higher activity which could be assigned to the less hindrance for ligand- receptor binding.Molecular modeling (docking) studiesAminopeptidase N (APN/CD13) which is assumed to be the target of ketoprofen-NGR conjugate is a zinc-dependent metalloprotease which is overexpressed in many disease such as cancer and inflammation. Bestatin is the first marketed APN inhibitor which has been introduced in 1976 (29). Therefor a docking study was conducted to investigate the possible binding mode of ketoprofen-spacer-NGR which has highest activity against HT-1080 cells. The result is presented in vβ3 integrin subunit that is overexpressed in cancer. In order to explore the possible mode of interaction for keto-RGD conjugate, docking study was carried out using 1L5G crystallography of αvβ3 integrin and the results are presented in 2 of asparagin moiety. The distance of Mn2+ ion in active site of enzyme from the carbonyl group of ketoprofen is 6.6A˚. Another Mn2+ ion in active site has 5.45A˚ distance from the carbonyl group of RGD peptide chain. Hydrophobic residue of (β)-Ala 218 with the distance of 3.9A˚ and 3.7A˚ has come into contact with ketoprofen rings. Hydroxyl group of (α)-Tyr178 forms hydrogen bond with NH of guanidine in keto-RGD with the distance of 3.88A˚ and also tyrosin ring has 3.49A˚ distance from CH2 side chain of Arg in keto-RGD.RGD target αRGD and NGR peptide sequences are capable of guiding a chemotherapeutic agent to its target. This strategy could be used for maximizing the efficiency of chemotherapy based on the fact that in many cancer cells, specific membrane receptors are overexpressed on the cell surface. The result of present study show that ketoprofen conjugated to RGD and NGR have higher cytotoxic activity compared to ketoprofen itself which strongly supports the hypothesis of targeting by peptide-drug conjugates.


The extent of genetic and epigenetic diversity between and within patient tumors is being mapped in ever more detail. It is clear that cancer is an evolutionary process in which tumor cell intrinsic and extrinsic forces shape clonal selection. The pre-clinical oncology pipeline uses model systems of human cancer – including mouse models, cell lines, patient-derived organoids and patient-derived xenografts – to study tumor biology and assess the efficacy of putative therapeutic agents. Model systems cannot completely replicate the environment of human tumors and, even within the same cancer model, data are often irreproducible between laboratories. One hypothesis is that ongoing evolutionary processes remain relevant in laboratory models, leading to divergence over time. In a recent edition of Nature, Ben-David and colleagues showed that different stocks of widely used cancer cell lines – a staple of cancer research over many decades – are highly heterogeneous in terms of their genetics, transcriptomics and responses to therapies. The authors find compelling evidence of positive selection based on ongoing mutational processes and chromosomal instability. Thus, the origin, culture conditions and cumulative number of population doublings of cell lines likely influence experimental outcomes. Here, we summarize the key findings of this important study and discuss the practical implications of this work for researchers using cell lines in the laboratory. Summary: A recent study in Nature demonstrated that ongoing mutational processes cause significant heterogeneity of widely used cancer cell lines. In this Editorial, the authors discuss the implications of these findings for the cancer and cell biology fields. To study human cancer biology, researchers can use patient biopsies or tissue from the surgical resection of tumors. However, this tissue represents a single snapshot in the life history of that tumor, typically at an advanced stage. In order to investigate the dynamic genetic and epigenetic course of cancer initiation, progression and metastasis, experimental model systems are required. More recent systems include genetically engineered mouse models (GEMMs) , patientin vivo, but are limited by species differences in oncogenic pathogenesis, the shorter lifespan of mice and, often, by the artificial, simultaneous introduction of multiple oncogenic driver events. Cell lines have the advantage of being derived from patients and are more easily manipulated in the laboratory. Nevertheless, the cancer research community has long appreciated that cell lines do not completely recapitulate human disease  were present in one data set but not the other. This discordance prompted the authors to examine the genetic variability within the cell lines in detail.https://portals.broadinstitute.org/ccle) and Connectivity Map (https://clue.io/cmap)]. The MCF7 strains also demonstrated substantial heterogeneity in their mutational status, chromosomal rearrangements and CNAs. Deep, targeted sequencing of 447 genes revealed that just 35% of coding non-synonymous single-nucleotide variants (SNVs) and insertions and deletions were shared by all of the strains, while 29% were unique to a single substrain. Even when specific mutations were present in all strains, the allelic fraction, defined as the number of variant reads divided by the total number of reads at a specific genomic locus, varied substantially. The authors highlight an activating PIK3CA mutation (G1633A), for which the allelic fraction fluctuated between 0.21 and 0.7, as an example likely to have functional consequences. Differences in larger genomic rearrangements, such as arm-level copy number events, and differential copy number gains and/or losses in hundreds of genes were also evident among strains. Of those events, only 13% of gains and 21% of losses were present across all strains, with 7% of gains and 13% of losses being unique to a single strain. Importantly, these events affected genes such as TP53, EGFR and PTEN, which are known to be gained or lost during breast cancer pathogenesis. This indicated that individual existing strains of the MCF7 cell line might be more biologically relevant to the study of estrogen receptor-positive breast cancer than others.Laboratories around the world maintain different substrains of cell lines, so the authors sourced 27 independent vials of MCF7, an estrogen receptor-positive breast cancer cell line, predominantly from two major Broad Institute projects [Cancer Cell Line Encyclopedia https://portals.broadinstitute.org/ccle) or the Sanger Institute's Cell Lines Project (https://cancer.sanger.ac.uk/cell_lines)] should be used for broad direction but specific mutation/pathway status should be confirmed locally.For existing cell lines, each laboratory's strain should be profiled. Reference genomes for cell lines [e.g. the Broad Institute's Cancer Cell Line Encyclopedia ((2)On adoption of new cell lines, identical cryovials of very early passage cell lines should be established on arrival and later passage working stocks derived from just one of these. Master stocks could then be used for validation of key findings prior to publication.(3)More rigorous monitoring and reporting of population doublings should become the norm, and substitute the current ‘passage number’ convention, which is imprecise. Cell lines should preferably be used within a defined window to minimize genetic drift.(4)The optimization, agreement and implementation of international ‘standard’ culture conditions for individual cell lines together with the documentation of heterogeneity metrics (e.g. measures of genomic divergence per passage in those conditions) in datasheets.(5)The reassessment of genomic landscape after significant culture bottlenecks induced by selection and/or single cell cloning (e.g. in CRISPR-Cas9 gene editing experiments).We believe that these data signal an end to the era in which researchers informally acknowledge that different strains of cancer cell lines are heterogeneous and unstable over long-term passage, but in practice treat them as though they were clonal entities. There are already established measures to minimize irreproducibility between laboratories, such as cell line identification , mycoplain vitro or in vivo, presents opportunities for researchers to improve the reporting and reproducibility of pre-clinical cancer research.In summary, Ben-David et al. reveal the scale of heterogeneity between strains of cancer cell lines and the ongoing nature of the processes that underlie this heterogeneity. Importantly, these processes are unlikely to be similar to the ongoing genomic processes in human tumors, and mean that as cell lines – already an idiosyncratic representation of a single patient's cancer – are expanded long-term, they become even less representative of human disease. Importantly, recognizing the variability within laboratory cancer models, whether


Picornaviridae, have been shown to have anticancer activity. Picornaviruses are small icosahedral non-enveloped, positive-sense, single-stranded RNA viruses infecting a wide range of hosts. They possess several advantages for development for cancer therapy: Their genomes do not integrate into host chromosomes, do not encode oncogenes, and are easily manipulated as cDNA. This review focuses on the picornaviruses investigated for anticancer potential and the mechanisms that underpin this specificity.Oncolytic viruses (OVs) form a group of novel anticancer therapeutic agents which selectively infect and lyse cancer cells. Members of several viral families, including  Herpes simplex virus-1 has shown increased efficacy when combined with cisplatin [Cancer remains one of the leading causes of death worldwide. Globally, there were around 18 million new cases of cancer in 2018, with up to 9.5 million deaths [isplatin . Oncolytisplatin ,5. Theseisplatin , Adenoviisplatin  and Mumpisplatin , and theHerpes simplex virus-1, also known as Talimogene Laherparepvec (T-VEC), by the US Food and Drug Administration (FDA) for the treatment of malignant melanomas [This was the approval of the first oncolytic virus, modified elanomas . In addielanomas ,11. Whilelanomas . This iselanomas . They alelanomas . ®), Vaccinia virus engineered to express GM-CSF with thymidine kinase deleted (PexaVec®), Measles virus engineered to express thyroidal sodium iodide importer (MV-NIS®) and naturally occurring Reovirus (Reolysin®). These viruses vary greatly with respect to several characteristics, including the species, genera, etc. to which they belong, their progress through clinical trials, cancer types they are used to treat and degree to which they have been engineered. Despite the inherent challenges presented in producing safe and effective oncolytic viruses, there are a number of naturally occurring and engineered viruses that have entered clinical trials. Included in their ranks are Adenovirus engineered to express granulocyte macrophage colony-stimulating factor (GM-CSF) , registered as CAVATAK . The onc3 TCID50 CVA21 (TCID—Tissue culture Infective Dose) and remained tumour-free until the end of the trial at day 35 [Potential oncolysis of a panel of six melanoma cell lines was inferred by increased levels of ICAM-1 and DAF . This wat day 35 . CVA21 wt day 35 ,39. Recet day 35 . Anothert day 35 . This reHigh ICAM-1 and DAF expression was confirmed in three multiple myeloma (MM) cell lines, with lower expression on peripheral mononuclear blood cells (PMBCs). MM cell lines showed cytopathic effect (CPE) on infection with CVA21 at a MOI as low as 0.2, increasing viral tires 100 to 1000-fold after 24 h. By contrast, CVA21 infection of PMBCs showed only modest CPE at MOI of 20 . When tr7 TCID50 CVA21 with a decrease in treatment tumour mass of 17 ± 6% compared to an increase in tumour mass of 2422 ± 932% in controls 26 days post-intervention [CVA21 was investigated for oncolytic activity against breast cancer . Of ninervention . CVA21 wrvention .To evaluate the effect of CVA21 on bladder cancers, a panel of ten bladder cancer cell lines were classified based on susceptibility . Six celThe combination of CVA21 with four common chemotherapy agents was assessed on a panel including breast cancer, colorectal cancer, pancreatic cancer and healthy cell lines . These eIn addition to native virus, the RNA of CVA21 transcribed from plasmid DNA has been shown to be cytotoxic on cell lines including hepatocellular carcinoma, lung cancer, rhabdomyosarcoma, and melanoma . This wa7, 108 or 109 TCID50 CAVATAK were safe, and five out of nine patients showed transient or stable tumour mass reduction, or stabilisation of tumours. In a Phase II clinical trial of CAVATAK in late stage melanoma (CALM), 57 patients were treated with up to 3 × 108 TCID50 CAVATAK intratumourally on days 1, 3, 5, 8 and 22, followed by injections once every three weeks for 18 weeks. This treatment regime resulted in no NCI CTCAE  grade 3 or 4 events, with 38.6% of patients showing immune-related progression-free survival at six months, and 19.3% of patients showing durable objective responses [The oncolytic activity of CVA21 in a diverse range of cancers both in vitro and in vivo prompted several clinical trials . A Phaseesponses . In an eesponses .8 TCID50), medium  and high  [In a Phase I/II clinical trial on systemic treatment of resistant malignancies (STORM), the enrolled patients with late-stage cancer were grouped into three CAVATAK dose cohorts, low  ,47. The  TCID50) .8 TCID50, 3 × 108 TCID50, or one of each. The second part of the study tested the combination therapy of two doses of 3 × 108 TCID50 CAVATAK with 10 mg MC. The study established the safety of CAVATAK and MC and recorded viral replication within tumours, complete tumour response and induced inflammation and apoptotic cell death [Revisiting the study of CVA21 with mitomycin C (MC) against bladder cancer cell lines, a Phase I/II clinical trial of CAVATAK in non-invasive bladder cancers (CANON study) was undertaken . The firll death .CVA21/CAVATAK as an oncolytic agent has garnered the greatest amount of research among the coxsackieviruses. However, the infection of CVA21 in humans is common, and pre-existing adaptive immunity to this strain could result in premature clearing of the virus before being able to exert its therapeutic effect on tumours. This prompted an evaluation of alternatives to CVA21 with less prevalent rates of immunity.Coxsackievirus serotypes A13, A15 and A18  have all been shown to infect and productively replicate in multiple melanoma cell lines, while not infecting peripheral blood mononuclear cells (PMBCs) . A studyOf 28 enteroviral strains tested against 12 human cancer cell lines and a bone marrow stroma control cell line, Coxsackievirus strains B2 Ohio-1 (CVB2 Ohio-1), B3 Nancy (CVB3 Nancy) and B4 JBV (CVB4 JBV) stood out as causing cell death in A549 and LK-87 lung cancer cells with no toxicity in controls . In a seN- and 6-O-sulfated heparan sulfate (HS) as a cellular receptor [Four strains of Coxsackievirus B3  were tested against colorectal cell lines expressing varying levels of CAR and DAF . Of the receptor . HS is areceptor . CVB3 Nareceptor .Three endometrial cancer (EC) cell lines, Ishikawa, HEC-1-A and HEC-1-B, were shown to express the cellular receptors for CVB3. When infected by CVB3 2035A at an MOI of 10, there was extensive lytic destruction of all three cell lines and high titres of viral progeny produced after 48 h . Mouse sFinally, Coxsackievirus B6 (CVB6) in the form of three non-pathogenic live enterovirus vaccine strains  was tested against healthy and cancer cell lines. LEV15 stood out as having very little infectivity in untransformed cell lines and varying infectivity in the cancer cell lines tested . LEV15 wPoliovirus is the causative agent of poliomyelitis, a disease infamous for causing life-long disability and paralysis throughout human history. Infection with Poliovirus (PV) is usually asymptomatic or manifested as transient flu-like symptoms and/or gastroenteritis . An esti6 TCID50 LAPV were sufficient to control tumour growth by 40% and induced cancer cells apoptosis, as shown by terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assay [Live Attenuated Poliovirus A vaccine containing the Sabin 1 strain (LAPV) has been tested for oncolytic activity in six human bone and soft tissue cancer cell lines expressing moderate to high levels of CD155, as well as one murine osteosarcoma . All butL) assay .Vaccinia virus vector that provides the P1 region [One way to attenuate PV neurotoxicity is by using replication incompetent Poliovirus 1 (PV1) replicons. Replicons are constructed by deleting the P1 region that encodes the capsid proteins . Once in1 region . PV1 rep1 region .133Gmono-cre illustrates another strategy to neuro-attenuate oncolytic PV [133G, was present in both cases, and so A133Gmono-CRE PV was isolated and used in further studies. Along with pre-exposure to mono-CRE PV, four intratumoural injections of A133Gmono-CRE PV into neuro-2a xenograft tumours were sufficient to clear tumours in 82% of mice, and also effectively vaccinate against re-challenge with neuro-2a cells [133Gmono-CRE PV cultures also achieved this effect [The PV mutant Alytic PV . Initiallytic PV . The ratlytic PV . In vitr2a cells . CD8+ T-s effect . These rBy far the most promising prospect for the use of PV in a cancer therapeutic setting is the chimeric virus, PVSRIPO. PVSRIPO ”) is the Sabin strain of PV1 (PVS) with IRES replaced with that of human Rhinovirus 2 (HRV2) ,82. The The basis for neuronal attenuation of PVSRIPO is multifactorial. Wild-type polioviral IRES binds the eIF4G:4A:4B complex (eukaryotic initiation factor 4G:4A:4B), unwinding the RNA, exposing the start codon and binding eIF3, while the HRV2 IRES does not in neuron lineage cells . PVSRIPOPVSRIPO has demonstrated potent oncolytic activity in a wide range of cancer cell lines derived from gliomas , breast Infection of ex vivo glioma primary cultures derived from stage II–IV patients with PVSRIPO showed drastic CPE by 6 h and complete CPE by 12 h, resulting in viral titres comparable to those produced by established glioma cell lines . The ex 7 plaque formation units (PFU) or a high dose of 109 PFU of PVSRIPO. In the intracranial model, the distinction between the administered doses of PVSRIPO was more pronounced, with the low dose extending median survival times by 122% and the high dose increasing median survival times by 153% compared to controls [CD155 expression in human breast cancer cells is relatively low . Howevercontrols .7 PFU) or high dose (109 PFU) PVSRIPO. While there was no significant dose response relationship between the two dose cohorts, median survival compared to controls increased by 187.5% and 200%, respectively, with one of ten mice in the low dose cohort, and three of ten mice in the high dose cohort surviving to the end of the trial [8 TCID50 PVSRIPO in both tumours showed an average of a 45% decrease in tumour mass by day 10, nearly clearing the xenografts by day 28. Tumours isolated from mice sacrificed at day 10 and 28 showed a temporal progression towards tumour death, with day 10 tumours displaying a dense core of intact cancer cells surrounded by a loose mantle of dispersed tumour cell foci and immune infiltrates. At day 28, the tumours were composed primarily of scar tissue, with minor foci of heterogenous composition [PVSRIPO was also tested in a glioblastoma mouse model . Athymiche trial . The acthe trial . Athymicposition . PVSRIPOposition .In melanoma cells, PVSRIPO effectively evades the innate immune response, where other related viruses such as Enchephalomyocarditis virus (EMCV) cannot, and the mechanism of this is independent of melanoma differentiation-associated protein 5 (MDA5) and the mitochondrial antiviral signalling protein (MAVS) .In vitro studies with SUM149 breast cancer cells and DU145 prostate cancer cells showed PVSRIPO had potent activity against these cell lines and that their infection released pro-inflammatory cytokines . By day PVSRIPO has been shown to infect human THP1 macrophages, but unlike many of the cancer cells studied, PVSRIPO infection of macrophages is sublethal and serves to induce expression of major histocompatibility complex class II and costimulatory molecules and further leads to (interferon-β) IFN-β, (interleukin-12) IL-12 and (tumor necrosis factor- α)TNF-α production . The imm7, 109 or 5 × 109 TCID5 PVSRIPO showed no side-effects, and all survived to the end of the trial [7 TCID50 [Following the recommendations of the World Health Organisation (WHO) for putative Poliovirus vaccines, before starting human trials, PVSRIPO had to be tested in primates. Ex vivo cultures of macaque and human kidney cells showed no species-specific infectivity for either PVS or PVSRIPO . Macaquehe trial . The virhe trial . These rhe trial ,94. A Ph7 TCID50 . In addi7 TCID50 .Enteric cytopathic human orphan (ECHO) viruses are part of the species Enterovirus B. Several echoviruses have been evaluated for selective tropism towards different cancers, including EV7, EV1, EV5, EV12, EV15, EV17, EV26 and EV29.Most abundant information regarding clinical applications is available on Echovirus-7 (EV7), registered by the name Rigvir . Rigvir Rigvir has shown toxicity in vitro against a broad range of cell lines, including melanoma, pancreatic adenocarcinoma, muscle rhabdomyosarcoma, mesenchymal stem cells, gastric carcinoma, lung carcinoma and human normal dermal fibroblasts .Clinical trials are referred to in the literature to have occurred between 1965 and 1991, but the results from these clinical trials are not readily available [One such retrospective study showed a statistically significant increase in 3-year survival of melanoma patients when treated with Rigvir post-surgery than surgery alone or with other immunomodulators . For besIn recent times, case studies of Rigvir treatment in a diverse range of cancers have been emerging in the literature. One such study followed a stage IV melanoma, stage IIIA small-cell lung cancer and stage IV hystiocytic sarcoma on a long-term Rigvir treatment . FollowiRigvir as a monotherapy and in combination with other therapies has also been tested in patients for whom the prognosis is especially poor. One patient with a basal cell carcinoma, a diagnosis with a median expected survival of five months, was treated with Rigvir post-surgery . The treα2β1, has been identified as the cellular receptor for EV1 [α2β1. This effect could be ablated by treatment with anti-α2β1 antibodies [Other Echovirus species have also been investigated for activity against cancer. Echovirus 1 (EV1) has been tested for activity against ovarian, prostate and gastric cancer cell lines. VLA-2, also known as integrin  for EV1 . Consisttibodies . EV1 wastibodies .α2β1, EV1 selectively destroyed the cancer cell lines [3 diameter was sufficient to reduce them to an average of 1 mm3 [3 and 107 TCID50 were used to test the efficacy of EV1 in large LNCaP tumours of 300 mm3. The low dose of EV1 had a transient but significant protective effect, while the mid- and high doses of virus showed lasting significant tumour suppression [While several prostate cancer cell lines as well as healthy prostate cell lines were also shown to express ll lines . Moreoveof 1 mm3 . Differepression .α2β1 was investigated [−6 to 3 × 10−2 TCID50/cell in MKN-45, AGS and NCI-N87 and >1 TCID50/cell in Hs746T. While Hs746T infection in vitro produced only minimal CPE and viral replication after 72 h, the other three cell lines produced a 100-fold increase in EV1 viral titre after 12 h [3 TCID50) responsible for a 47.38% reduction, the medium dose (105 TCID50) for a 68.5% reduction and a 71.5% reduction in the high dose (107 TCID50) cohort. For reference, mice treated with PBS had an average flux increase of 28.75%, and most were sacrificed due to disease progression. Interestingly, circulating virus was initially different across dose groups, but then equalised [The minimum infectious dose of EV1 to produce CPE in four gastric cancer cell lines expressing stigated . These wter 12 h . MKN-45 qualised .Echovirus 5 (EV5) represents an interesting study into the oncolytic activity of the wild-type virus genome and the naked RNA transcribed from a cDNA clone . EV5 wasFinally, the broader oncolytic ability of Echoviruses was tested in a study with viruses including EV12, EV15, EV17, EV26, and EV29 against a selection of six colorectal cell lines . EV12 atBovine Enteroviruses are endemic in cattle, being routinely isolated from cow faeces, and are not pathogenic to humans . Bovine Vibrio cholera neuraminidase (VCN) showed a dose- and time-dependent inverse correlation between VCN exposure and viral titre following BEV 261 infection in a panel of susceptible cancer cells [BEV1 was first investigated for antitumour activity in 1971, wherein 65% of cancer cell lines met the threshold for significant viral killing by BEV1, compared to 4% of healthy cell lines . The perer cells .Interestingly, whereas other picornaviruses can bio-adapt to cell lines (see: Echovirus 7), this could not be achieved with BEV1 . The samIn addition to murine studies, BEV1 was also tested in rabbits and dogs and shown to be well tolerated . Mice beSeneca Valley Virus (SVV) was originally discovered as a contaminant of cell culture . The sel8 viral particles per kilogram (vp/kg) SVV could clear pre-established H446 xenograft tumours and most mice treated with 107 vp/kg cleared tumours, with two exceptions. Similarly, when athymic mice with Y79 retinoblastoma xenograft tumours were treated with 108, 1011 or 1014 vp/kg SVV, six of eight, seven of seven and five of seven mice, respectively, cleared the tumours [The oncolytic potential of SVV was first highlighted in a 2007 study . Screeni tumours . This tr13 vp/kg SVV [Concurrently, SVV was shown to have activity against Weri retinoblastoma cells in vitro . Doses ap/kg SVV .12 vp/kg. SVV treatment increased the survival times in mice of both tumour sizes. Of note is that SVV could cross the blood–brain barrier (BBB) and did not infect any healthy mouse brain cells [To research SVV in the application against medulloblastoma (MB), mouse MB models were established using ex vivo primary cultures isolated from 10 paediatric patients . Primaryin cells .50  values of 1.6 × 10−3 vp/cell, 3.1 × 10−4 vp/cell and 3.9 × 10−3 vp/cell, respectively. Gene expression profiles of permissive and refractory cell lines showed that higher expression of late neurogenic transcription factor NEUROD1 and lower expression of the early neurogenic transcription factor ASCL1 correlated with permissivity to SVV [9 to 1011 vp/kg SVV, and two out of six mice completely cleared tumours when treated with 1014 vp/kg SVV. Kinetics studies showed peak replication of SVV occurred at day three [Small-cell lung cancer (SCLC) is an aggressive type of tumour. There are two SCLC groups: classic and variant, which make up for 70% and 30%, respectively [Since SVV could exert a therapeutic effect in medulloblastoma models, it raised the question of its applicability in other cancers for which the BBB presents a significant obstacle, such as gliomas. In a study of six ex vivo glioma primary cultures, four were found permissive to SVV infection at an MOI as low as 0.5, while the other two remained resistant up to an MOI of 25 . Replica12 vp/kg NTX-010 were seen in neuroblastoma, rhabdomyosarcoma, rhabdoid tumour, Wilms tumour and glioblastoma cell lines [SVV-001  is registered by Neotropix inc. as NTX-010 . A preclll lines . Taken t7 to 1011 vp/kg showed no dose-limiting toxicities, with flu-like symptoms mainly manifesting in the lower dose cohorts. Neutralising antibodies were detected as early as two weeks into treatment. In terms of outcomes, one patient showed disease stabilisation, while another five had minor responses, which were not sufficient to meet RECIST criteria  [9, 1010 or 1011 vp/kg NTX-010, and in part B, patients were given oral (days 1 to 14) and intravenous (days 1 and 8) cyclophosphamide, in combination with two doses of 1011 vp/kg NTX-010. The study showed that NTX-010 was well tolerated with a single dose-limiting toxicity event recorded. While no objective response was observed, six patients did show disease stabilisation. A rapid neutralising antibody response to NTX-010 was also shown in this trial [11 vp/kg NTX-010, or Arm B, the saline control. Grade 4 adverse events were seen in three of the Arm A patients and none of the Arm B. Between the two cohorts, there was no difference in progression-free survival, and overall survival was slightly less in Arm A (83%) vs. Arm B (85%). The trial was also prematurely terminated to investigate the death of one patient, which was confirmed to be unrelated to NTX-010 treatment [First, a phase I clinical trial of 30 patients with SCLC was undertaken . Patienttumours) . The pattumours) . Shortlytumours) . In partis trial . Finallyis trial . A cohorreatment . It is ireatment . Based oTheiler’s murine encephalomyelitis virus (TMEV) is a natural murine pathogen that has been engineered to express tumour antigens to induce a targeted antitumour immune response and thus act as a cancer vaccine.TMEV engineered to express chicken ovalbumin (TMEV–OVA) can generate host immune responses against OVA-expressing tumours ,120. The121–130) [121–130 region has been attempted in order to direct a greater proportion of the cytotoxic immune response to expressed foreign antigen. Deletion of this immunogenic region is shown to increase the relative amount of CTLs primed against foreign antigen [During the host immune response generated to TMEV expressing foreign antigen, the adaptive immune response targets a combination of the foreign antigen and TMEV antigens. TMEV is known to harbour a highly immunogenic region in the VP2 structural protein (VP2121–130) . Suppres antigen . Vector  antigen . However antigen . In fact antigen .Wild-type DA strain of TMEV has been investigated in vivo against breast cancer and melanoma cell tumours, without a significant protective effect in either . Wild-tyEncephalomyocarditis virus (EMCV) was discovered in 1945 when a captive male gibbon suddenly and inexplicably died of pulmonary oedema and myocarditis . Mice trAs far back as 1965, EMCV, referred to as Columbia-SK virus, showed oncolytic activity against fructose sarcomas in mice and rats, with a more pronounced effect in rats . The intVHL is a tumour suppressor protein with a role in tumour necrosis factor (TNF)-α mediated cell apoptosis  and oxyg24, a poly-C truncated attenuated MEV, demonstrated oncolytic activity in a miscellaneous panel of cancer cell lines that included several multiple myeloma cells of human and murine origin [24 caused significant CPE in the majority of cancer cell lines tested [24 infection. BALB/c mice bearing MPC-11 tumours were treated with either a single intravenous (IV) injection of 107 TCID50 vMC24 or intratumoural (IT) injection of 106 TCID50 vMC24. While IV injection showed only minimal tumour suppression, IT administration of vMC24 resulted in a decreased tumour volume within five days. However, three mice in the IV group and two in the IT group developed paralysis, with thee more dying overnight, suggestive of a heart failure [24 showed potential as oncolytic agent but with lethal toxicity. This prompted a search for novel strategies to attenuate vMC24, such as the incorporation into the viral genome of sequences complementary to healthy tissue specific micro-RNAs [24 variants with complementary micro-RNA sequences (miRT viruses) were generated. The miRT viruses had combinations of miR125b (expressed in brain tissue), miR124 (expressed in neurons), miR133 and miR208a (enriched in cardiomyoctyes). Among these, a construct labelled vMC24-NC stood out as having very low viral titres in the brain, spine and heart of experimentally infected C57BL/6 mice. vMC24-NC had two miR124 sequences in the 5′ UTR, and one miR133b and one miR208a in the 3′ UTR. In vivo experiments with mice bearing MPC-11 tumours showed that four of ten mice treated with vMC24-NC cleared tumours and two delayed tumour growth, with no negative side-effects. The mice treated with vMC24 had a rapid tumour regression, but seven out of ten developed paralysis. Furthermore, vMC24-NC was also shown to eliminate tumours in a dose-dependent manner while remaining genetically stable in vivo [Similar to Poliovirus, the research into oncolytic Mengovirus (MEV) focused around novel attenuation methods . A studye origin . vMC24 cs tested . Of the  failure . Therefocro-RNAs ,128. Mul in vivo .This review discussed picornavirus species investigated for anticancer activity . These v


Additionally, high extracellular lactate cultures were labelled in parallel with [U-13C] lactate, which provided qualitative information regarding the lactate uptake and metabolism. The 13C-MFA model, which incorporated the measured extracellular fluxes and the parallel labeling mass isotopomer distributions (MIDs) for five glycolysis, four tricarboxylic acid cycle (TCA), and three intracellular amino acid metabolites, predicted lower glycolysis fluxes in the high lactate cultures. All three cell lines experienced reductive carboxylation of glutamine to citrate in the TCA cycle as a result of high extracellular lactate. Reductive carboxylation previously has been observed under hypoxia and other mitochondrial stresses, whereas these cultures were grown aerobically. In addition, this is the first study to investigate the intracellular metabolic responses of different stages of breast cancer progression to high lactate exposure. These results provide insight into the role lactate accumulation has on metabolic reaction distributions in the different disease cell types while the cells are still proliferating in lactate concentrations that do not significantly decrease exponential growth rates.In cancer tumors, lactate accumulation was initially attributed to high glucose consumption associated with the Warburg Effect. Now it is evident that lactate can also serve as an energy source in cancer cell metabolism. Additionally, lactate has been shown to promote metastasis, generate gene expression patterns in cancer cells consistent with “cancer stem cell” phenotypes, and result in treatment resistant tumors. Therefore, the goal of this work was to quantify the impact of lactate on metabolism in three breast cell lines , in order to better understand the role lactate may have in different disease cell types. Parallel labeling metabolic flux analysis ( For example, when lactate was injected into mice with xenografts of the human breast cancer cell line MDA-MB-231, metastasis increased ten-fold  glucose, [U-13C] L-glutamine, and [U-13C] sodium lactate, where the control cultures were only labeled with  glucose and [U-13C] L-glutamine. The lactate labeling data for the high-lactate cultures provided uptake and direct intermediate tricarboxylic acid (TCA) cycle labeling information. The intracellular metabolic fluxes were predicted for each cell line and each condition from the glucose and glutamine labeling data.The aim of this study was to determine the role of extracellular lactate on the metabolism of three different proliferating breast cell lines . Each cell line was grown in a low glucose control  media or in a high-lactate media, where sodium lactate was added to the control medium to make the high-lactate medium. The high-lactate concentration was selected for each cell line such that equivalent exponential growth rates for the control and high-lactate cultures were maintained. These equivalent exponential growth rates were important for the modeling assumptions for two reasons: 1) the quasi-steady state assumption was met equally well by both conditions, and 2) it allowed for normalizations of fluxes between the conditions for the same cell line based on biomass generation. Parallel labeling experiments were conducted with  glucose, [U-13C] L-glutamine, and [U-13C] sodium lactate, all purchased from Sigma-Aldrich.MCF 10A (ATCC CRL-10317), MCF7 (ATCC HTB-22) and MDA-MB-231 (ATCC HTB-26) cells were from the American Tissue Culture Collection (ATCC). MCF 10A cells are a non-tumorigenic breast cell line, MCF7 cells are a tumorigenic, luminal breast cancer cell line, and MDA-MB-231 cells are a metastatic, basal breast cancer cell line . Dulbecc4 cells/cm2 into either 6-well plates or T-25 flasks in the control growth media. Cells were cultured at 37°C in a 5% CO2 humidified incubator. After 24-h, the media for the MDA-MB-231 cultures was exchanged and the experimental conditions were introduced. The media exchange time is used to set time 0 for all cell lines. For MCF 10A and MCF7 cell lines the lag phase was longer, so the media was replenished at 24-h post seeding. The media was exchanged 48-h post seeding for MCF10A and MCF 7 to introduce the experimental conditions. For clarity, the full experimental setup is shown in 13C] glucose) and glutamine tracers (95% molar enriched [U-13C] L-glutamine) were performed for the control cultures, whereas the high-lactate cultures also had parallel replicates for the lactate tracer (50% molar enriched [U-13C] sodium L-lactate). Six-well plates (Nunc) were used to obtain cell numbers and glucose and lactate concentrations with six replicates for each time point. T-25 flasks were used to obtain extracellular amino acid concentrations and intracellular MID measurements in triplicate.All cell lines were seeded at 2 x 10Cell numbers were obtained using the Scepter 2.0 Handheld Automated Cell Counter (Millipore). Glucose and lactate concentrations were measured using a YSI 2700 Bioanalyzer.13C] algal amino acid solution, which were then used to quantify the amino acid concentrations in the extracellular medium.Intracellular and extracellular metabolites and specific metabolite fragments used for identification were extracted and derivatized according to the protocol outlined by Ahn and Antoniewicz , with a GC-MS analysis was performed using a Hewlett Packard 7683 GC equipped with an HP-5  capillary column, interfaced with a Hewlett Packard 5973 MS operating under ionization by electron impact as 70 eV and 200°C ion source temperature. The injection port and interface temperatures were both 250°C while helium flow was maintained at 1 mL/min. Mass spectra were recorded in full scan mode for amino acid quantification and in single ion mode (SIM) for MIDs as well as internal standards and standardization curves. MIDs were obtained by integration of single ion chromatograms and corrected for natural isotope abundances using the Metran software , 32.-1  glucose included in the MFA simulations were 3-phosphoglycerate (3PG), dihydroxyacetone phosphate (DHAP), pyruvate, lactate, and alanine. The MIDs metabolites labeled by [U-13C] glutamine included in the MFA simulations were succinate, malate, α-ketoglutarate (AKG), glutamate, citrate, glutamine, and pyruvate. The MIDs of metabolites labeled by [U-13C] lactate were not included in the MFA simulations.e Metran , which ue Metran . Metaboltal data , 36. In tal data , 21. Mosμ is the growth rate (h−1). C48 and C24 are the individual metabolite concentrations (mM) at 48 and 24 hours after the media exchange, respectively. X24 is the cell concentration (cells/mL) at 24 hours after the media exchange. In Meadows et al. (2008)  glutamine) pathways [13C] glucose, the percent isotope labeling for DHAP, 3PG, pyruvate, alanine, and lactate were 40% or higher for all three cell lines. No significant differences were observed for MIDs from glycolytic metabolites  between the three cell lines and between the control and high-lactate conditions. These results reflect the consistently high flux through glycolysis for each of the three cell lines and the equivalent growth rates between conditions within a given cell line. These results also confirm that the 24-h labeling time was sufficient to reach isotopic steady state for  glucose. The intracellular MID profiles for all three cell lines are included in Fig B in  glutamine. Using [U-13C] glutamine, the observed isotopomer labeling for intracellular glutamine, AKG and glutamate was over 70%. 13C] glutamine, as an example, for the TCA metabolites. Pyruvate and lactate were minimally enriched from the glutamine isotope across the cell lines and both conditions, where For the three cell lines and both conditions, glutamine, AKG, and glutamate were highly labeled from [U-13C] glutamine tracer. The MID profiles for these metabolites were similar for all three cell lines; however, citrate MIDs were significantly different between the control and high-lactate cultures within a cell line, indicating the effect of lactate was significant. More specifically, a significant increase in M5 citrate labeling was observed in the high-lactate conditions for all three cell lines Click here for additional data file.


Glioblastoma (GBM) is the most common and aggressive type of primary brain cancer. With median survival of less than 15 months, identification and validation of new GBM therapeutic targets is of critical importance.In this study we tested expression and performed pharmacological characterization of the calcitonin receptor (CTR) as well as other members of the calcitonin family of receptors in high-grade glioma (HGG) cell lines derived from individual patient tumours, cultured in defined conditions.1/2 and p38 MAP kinases, and Ca2+ mobilization, supportive of low cell surface receptor expression.Previous immunohistochemical data demonstrated CTR expression in GBM biopsies and we were able to confirm CALCR (gene encoding CTR) expression. However, as assessed by cAMP accumulation assay, only one of the studied cell lines expressed functional CTR, while the other cell lines have functional CGRP (CLR/RAMP1) receptors. The only CTR-expressing cell line (SB2b) showed modest coupling to the cAMP pathway and no activation of other known CTR signaling pathways, including ERKExome sequencing data failed to account for the discrepancy between functional data and expression on the cell lines that do not respond to calcitonin(s) with no deleterious non-synonymous polymorphisms detected, suggesting that other factors may be at play, such as alternative splicing or rapid constitutive receptor internalisation.This study shows that GPCR signaling can display significant variation depending on cellular system used, and effects seen in model recombinant cell lines or tumour cell lines are not always reproduced in a more physiologically relevant system and vice versa.The online version of this article (10.1186/s12885-019-5369-y) contains supplementary material, which is available to authorized users. Glioblastoma (GBM) is the most common and aggressive type of primary brain cancer , 2. GBM The calcitonin family of receptors consist of the calcitonin receptor  and the calcitonin receptor-like receptor ; both are class B (or secretin-like) G protein-coupled receptors (GPCRs). CTR and CLR can form complexes with the accessory single-transmembrane-domain proteins known as the receptor activity-modifying proteins (RAMPs)  generatiEMA/109665/2013). Furthermore, in mouse thymic lymphoma models, where CTR is expressed as an amylin receptor, amylin treatment leads to metabolic reprogramming (switch from glycolysis to oxidative phosphorylation) resulting in increased susceptibility to apoptosis  and with 1 mM DTT, 1 mM PMSF and 1x cOmplete Mini Protease Inhibitor Cocktail. Suspensions were kept on ice for 10 min to allow solubilisation of membrane proteins and centrifuged for 20 min at 16,300 g and 4 °C. The resulting supernatant (membrane protein fraction) was collected and all fractions stored at − 80 °C. 50 μg of BCA quantitated membrane fractions were denatured at room temperature in Laemmli sample buffer and subjected to electrophoresis on 8% gels and transfer to PVDF using standard methods. Membranes were probed with anti-CTR antibody  followed by HRP-conjugated secondary, enhanced chemi-lumiscence and detection with a LAS-3000 Imaging System .Cells were harvested by scraping into PBS containing 1x cOmplete Mini Protease Inhibitor Cocktail (Sigma-Aldrich) and 1 mM EDTA and subjected to small-scale subcellular protein fractionation to enrich for the membrane fraction. Briefly, cell suspensions were microfuged for 5 min at 350 g and 4 °C, supernatant discarded and cell pellet homogenised in 2.5 volumes of hypotonic buffer  for expression in primary tumours, cell lines and xenograft tumours of calcitonin receptor family genes along with calcitonin and amylin by considering all microarray data with a detection .05 Fig. . In cont.05 Fig. , possibl.05 Fig. . In cont.05 Fig. . We ther.05 Fig. a.Fig. 1MDue to the discrepancy between the microarray data and our initial western blot screen, cell lines WK1, JK2, SB2b and PB1 were selected for further analysis. We tested for mRNA expression using TaqMan qPCR, which has been reported as more reliable for detection of low expression GPCR transcripts . This sh50 = 9.4 ± 0.3) and hCT (pEC50 = 8.0 ± 0.2) were similar to the known affinities of these agonists for CTR. The similarities in affinity and cellular potency are consistent with low receptor expression and limited receptor reserve. We also observed a higher Emax for hCT (43% of forskolin) compared with sCT (35% of forskolin), consistent with higher efficacy of this lower affinity agonist [50 = 6.8 ± 0.5) is consistent with signalling via CTR in the absence of any RAMP [In a wide variety of recombinant and ex vivo settings CTR is most strongly coupled to the stimulatory hetero-trimeric Gα subunit, Gαs, that activates adenylate cyclase –47. CTR  agonist . The lowany RAMP  indicati50 = 5.9 ± 0.5) and PB1 (pEC50 = 6.1 ± 0.5) is inconsistent with an AMY receptor phenotype but is consistent with the CGRP (CLR/RAMP1) receptor, with application of 1 μM of the CGRP antagonist (CGRP(8–37)) abolishing this response   [iCa2+ mobilisation assay with sCT or amylin agonists in SB2b, WK1 or PB1 cell lines  . We were+ . We we50 = 8.6 ± 0.1 for PB1, pEC50 = 8.3 ± 0.2 for JK2 and pEC50 = 8.8 ± 0.1 for WK1) and with lower potency to adrenomedullin    [To understand what (if any) relationship CTR expression may have with tumour progression and patient outcome we extracted the raw expression data for CTR from two public databases (IVY-GAP  and TCGAM = 0.3) . For patM = 0.3)  reportedM = 0.3) . The onlM = 0.3) . To contM = 0.3) , we mappM = 0.3)  (AdditioM = 0.3) .1 receptors, albeit that the CTR was expressed at low levels. Neither of the CTR activating peptides, hCT or amylin, were expressed at levels above threshold, suggesting there is no autocrine production of these peptides by these GBM cell lines. No coding polymorphisms, apart from the well-characterised c-terminal tail leucine/proline polymorphism [Expression profiles revealed by RT qPCR indicate that each of the four model HGG cell lines investigated here generate mRNA encoding components of CTR, AMY, CGRP and AMmorphism , were demax than hCT. The molecular basis for the signalling profile observed in the SB2b GBM cell line is consistent with low or no receptor reserve but reveals apparent differences in efficacy, consistent with our data in recombinant systems [iCa2+ mobilization assay suggesting limited Gq coupling of CTR in the SB2b cell line. Additionally, we were not able to detect either pERK1/2 or p38 MAPK response in this cell consistent with no downstream effect of CTR activation on cellular metabolism or proliferation.Although, RT qPCR data show mRNA encoding multiple receptors of the CTR family in the SB2b cell line , functionally we were only able to confirm CTR. The potency with which CTR agonists elicit a cAMP response is consistent with a low level of endogenously expressed receptor. Weak responses to αCGRP, amylin and adrenomedullin can be attributed to CTR activation. In this cell line, sCT and hCT had distinct profiles with sCT demonstrating higher potency but lower E systems , and furiCa2+ assay (PB1 and WK1). However, we have previously shown that both JK2 and WK1 cell lines are susceptible to anti-CTR-immunotoxin  mediated cell killing [PB1, JK2 and WK1 cell lines  had detectable mRNA and western blot immuno-reactivity consistent with CTR expression . In spite of this we could not detect a functional CTR response as assessed by cAMP accumulation  and  killing . In othe killing , 55, sugDespite a lack of functional CTR in PB1, JK2 and WK1 cell-lines, all three displayed a potent cAMP accumulation response to αCGRP, a less potent response to adrenomedullin, with a very weak response to amylin. In addition, specific CGRP and AM receptor antagonists confirmed the presence of CLR/RAMP1 type CGRP and not an AM receptor in these cells. This is in agreement with RT qPCR gene expression data showing expression of both CLR and RAMP1. However, CGRP is unlikely to be tractable as a target for treating GBM given its broad expression in the brain. There was no clear correlation between pharmacological profiles and clinical classification of the originating GBM tumour subtype as SB2b and PB1, both of classical GBM subtype, had distinct pharmacological profiles of CTR and CLR/RAMP1 receptor.Taken together this data indicates that we have a rather incomplete understanding of CTR (and related receptor) function in certain (patho)-physiological circumstances. While the CTR is expressed in a significant subset of GBM tumours it may only be tractable as a target by leveraging the compromised blood brain barrier characteristic of these tumours while taking advantage of the rapid cycling of CTR to deliver a toxic payload.Our analysis of published CTR expression from IVY-GAP  or TCGA Additional file 1:Figure S2. MAP kinase response to sCT in SB2b cells and TCGA survival data. No detectable ERK1/2 phosphorylation (A) or p38 (B) in response to stimulation with 1 μM sCT in SB2b cell line while a robust response to 10% FBS is seen; Data are presented as mean + S.E.M. of 3 replicates of a representative experiment. (C) ERK1/2 Phosphorylation response in SB2b cell line was induced by 0.1% FBS. No suppression of the induced response after stimulation sCT or hCT was seen at the concentrations tested (C). Data are presented as mean + S.E.M. of 3 replicates of a representative experiment. (D) Log2 expression (FPKM) ofr CALCR transcript in patients with survival data from the TCGA database plotted as a scatter plot against survival. (PDF 915 kb)Additional file 2:Figure S1. cAMP and iCa2+ mobilization in response to CTR agonists. A, Characterization of cAMP accumulation (30 min) in WK1 cells in response to stimulation by sCT alone or in presence of 1 μM forskolin. Data are presented as mean + S.E.M. of 3 replicates of a representative experiment. Absence of intracellular calcium mobilization response to sCT and rAMY in WK1 (B), SB2b (C) and PB1(D) cell lines while maintaining robust response to 10 μM ATP and 1 μM ionomycin. Data are presented as peak values of response measured in relative fluorescence units. Data are presented as mean + or - S.E.M. of 3 replicates of a representative experiment. (PDF 907 kb)Additional file 3:Figure S3. Mapping reported CTR mutations to our a molecular model of the CTR [ the CTR . A, mutaAdditional file 4:Figure S4. Alignment of vertebrate CTR sequences. Alignment of a subset of validated and predicted CTR sequences from mammals and aves with reptile and amphibian sequences used as outgroups. Sequences were obtained from NCBI homologene filtering for reference sequences only. These were then manually curated and an alignment was performed using Clustalw Omega. Conserved asparagine (yellow) and cysteine (purple) residues in the N-terminus have been manually annotated and TMMHM used to predict TM helices which were manually curated and are indicated in blue. Putative LOF mutations are highlighted in red. (PDF 211 kb)


Fusarium solani strain, which is associated with the plant Aglaonema hookerianum Schott. FUS showed rapid and sub-micromolar IC50 against pancreatic cancer cell lines. Time-dependent survival analysis and microscopy imaging showed rapid morphological changes in cancer cell lines 4 h after incubation with FUS. This provides a new chemical scaffold that can be further developed to obtain more potent synthetic agents against pancreatic cancer.As a part of our ongoing research on endophytic fungi, we have isolated a sesterterpene mycotoxin, fusaproliferin (FUS), from a  Pancreatic adenocarcinoma is a leading cause of adult cancer mortality. It is presently untreatable, with a 5-year survival rate of ~5% . As earlFusarium solani strain, isolated from the plant Aglaonema hookerianum Schott. (Family: Araceae), was investigated. F. solani has been proved as a potent source of structurally-diverse natural compounds with cytotoxic activity, such as karuquinone A and karuquinone B + with a molecular formula of C27H40O5 , accounting for 8 degrees of unsaturation. The resonances at δ 170.9 and 207.9 ppm in the 13C-NMR spectrum were characteristic of the presence of two carbonyl carbons of an ester and a ketone, respectively. The 1H and 13C-NMR data, in conjunction with the DEPT-135 spectrum , three sp3 methines , three sp2 methines , one sp3 quaternary carbon (15-), five sp2 quaternary carbons , two carbonyl carbons of an ester (26-OCOCH3) and a ketone (16-CO-). The presence of three sp2 methines and five sp2 quaternary carbons, along with one each of an ester and carbonyl moiety, proved the presence of six double bonds in this compound, and thus indicated that it was a bicyclic compound. After deducting the acetyl moiety ‘OCOCH3’ , the compound consisted of 25 carbons, which indicated it as a sesterterpene.FUS was obtained as a white gum. The structure of FUS  was confl values . Accuratspectrum , proved 50, but the activity against the pancreatic cancer cell lines were notably better than the breast cancer cell lines. FUS was between 3 to 58 times more potent than gemcitabine in pancreatic cancer cell lines, but doxorubicin was superior against both breast cancer cell lines compared to FUS. The therapeutic utility of FUS was further investigated against WI38, a non-tumor lung fibroblast cell line. FUS was found to be cytotoxic against WI38 with a high micromolar IC50  (p < 0.03) and at 8 h (p < 0.01) compared to doxorubicin  a. Similaorubicin , althougThe morphological changes in the MIAPaCa2 cell line after incubating with FUS were monitored using a Nikon TS100 inverted microscope fitted with a camera. The cells appeared to show both apoptotic and necrotic damages within 4 h post-incubation, and the damages were fully evident at the 8-h time point c,d. ThesF. solani. This compound showed potent and rapid cytotoxicity against both pancreatic and breast cancer cell lines tested in this study. The complex structure and intriguing biological activity of FUS make it a good target for chemical synthesis and a lead structure for a medicinal chemistry project to develop a new anticancer drug against pancreatic cancer.In summary, FUS is a known sesterterpene mycotoxin isolated from the endophytic fungus A. hookerianum was collected from Pablakhali, Rangamati, Chittagong Hill tracts, Bangladesh on 10 August 2014 and identified by the taxonomist of Bangladesh National Herbarium, Mirpur, Dhaka. A voucher specimen of the plant has been deposited (Accession no.: DACB 40633) in the herbarium for further reference , and the data were processed by Thermo XCalibur 2.2. Column chromatography was carried out on silica gel . Organic solvents, potato dextrose agar (PDA) medium, and TLC plates were purchased from Merck, Germany.The NMR spectra were recorded on a Bruker 400 MHz NMR spectrometer using CDClA. hookerianum bearing the internal strain no. AHPE-3, AHPE-4 (Fusarium sp. (Fusarium sp. (AHPE-4), Colletotrichum sp. (AHLE-1) and Colletotrichum sp. (AHLE-4). The fungus AHPE-4 was selected for further investigation, based on the brine shrimp lethality bioassay data  was cut with a sterile scalpel and stored at 4 °C in a sterile polyethene bag prior to use. Endophytic fungi were isolated from the fresh plant parts following the procedure, established at Pharmaceutical Sciences Research Division, BCSIR Laboratories, Dhaka, Bangladesh ,12,13,14, AHPE-4 , AHLE-1 rium sp. .  and the intervening 5.8S rRNA region was amplified and sequenced using electrophoretic sequencing on an ABI 3730 × l DNA analyzer  using Big Dye Terminator v 3.1 cycle sequencing kit . The ITS regions of the fungus were amplified using PCR  and the universal ITS primers, ITS4 (5′-TCC GTA GGT GAA CCT GCG G-3′) and ITS5 (5′-GGA AGT AAA AGT CGT AAC AAG G-3′). The PCR products were purified and desalted using the Hot Start Green Master Mix  and sequenced on an ABI 3730 × l DNA analyzer . The sequences were aligned and prepared with the software Chromas  and matched against the nucleotide-nucleotide database (BLASTn) of the U.S. National Center for Biotechnology Information (NCBI) for final identification of the endophytic isolate. Finally, the sequence data (SI) were deposited in the Gen Bank database (accession number MG75792), which revealed 99% similarity other related fungal isolates of F. solani (AHPE-4), isolated from the petiole of the plant A. hookerianum, was cultivated at 28 ± 2 °C for 28 days on potato dextrose agar (PDA). The culture media were extracted with ethyl acetate for seven days in an air-tight, flat-bottom container with occasional shaking and stirring. This procedure was repeated three times to obtain the crude extract. The extract of endophytic fungi was then filtered using sterilized cotton filter followed by Whatman no. 1 filter papers. The solvent was evaporated with a rotary evaporator at low temperature (40 °C–50 °C) and reduced pressure. The fungus 2SO4 spray reagents. The column fraction of petroleum ether/15% ethyl acetate was subjected to preparative TLC on silica gel  to obtain FUS.The crude fungal extract (8 gm) was subjected to column chromatography for fractionation on silica gel (70–230 mesh) using gradients of petroleum ether/ethyl acetate, then ethyl acetate, followed by a gradient of ethyl acetate/methanol, and finally methanol, to afford a total of 15 fractions. These fractions were screened by TLC on silica gel under UV light and by spraying with vanillin-H1H-NMR, CDCl3): δ 2.40 , 1.74 , 5.27 , 2.30 , 2.06 , 2.30 , 2.11 , 5.15 , 2.11 , 1.82 , 1.82 , 1.65 , 4.08 , 5.40 , 2.40 , 1.95 , 2.69 , 2.80 , 1.66 , 1.66 , 1.59 , 1.02 , 4.31 , 4.27 , 1.33 , 2.06 , 5.56 . 13C-NMR: δC 39.1 (C-1), 121.4 (C-2), 138.2 (C-3), 40.3 (C-4), 23.8 (C-5), 124.3 (C-6), 132.9 (C-7), 34.9 (C-8), 29.7 (C-9), 76.5 (C-10), 136.5 (C-11), 128.9 (C-12), 28.7 (C-13), 49.6 (C-14), 49.0 (C-15), 207.9 (C-16), 147.3 (C-17), 146.7 (C-18), 33.7 (C-19), 15.5 (C-20), 15.3 (C-21), 10.4 (C-22), 16.2 (C-23), 66.4 (C-24), 14.5 (C-25), 170.9 (C-26), 20.9 (C-27). HRESIMS m/z 467.2778 [M + Na]+ .18 mg, white, amorphous sticky mass; , horse serum  and penicillin-streptomycin . The BXPC3 cell line was maintained in RPMI-1640 medium , supplemented with fetal bovine serum , and penicillin-streptomycin . The MDA MB 231 cell line was maintained in Dulbecco’s modified Eagle’s medium , supplemented with fetal bovine serum , l-glutamine , non-essential amino acids  and penicillin-streptomycin . The MCF7 cell line was maintained in Eagle’s Minimum Essential medium supplemented with fetal bovine serum , 0.01 mg/mL human recombinant insulin and penicillin-streptomycin . During seeding, cells were counted using a Neubauer hemocytometer  by microscopy  on a non-adherent suspension of cells that were washed in PBS, trypsinized, centrifuged at 8 °C at 8000 rpm for 5 min, and re-suspended in fresh medium.The MIA PaCa2 (pancreatic adenocarcinoma), BXPC3 (pancreatic adenocarcinoma), MDA-MB-231 (triple-negative breast cancer), MCF-7 (estrogen receptor positive breast cancer) cell lines were obtained from the American Type Culture Collection. The MIA PaCa2 cell line was maintained in Dulbecco’s modified Eagle’s medium , supplemented with fetal bovine serum (10% 2 humidified atmosphere using an appropriate medium. The cell count was adjusted to 105 cells/mL and 2500 cells (MDA-MB-231) or 5000 cells (A4 and WI-38) were added per well. The cells were incubated for 24 h, and 1 μL of the appropriate inhibitor concentrations was added to the wells in triplicate. After 96 h of continuous exposure to each compound, the cytotoxicity was determined using the 3--2,5-diphenyltetrazolium bromide (MTT)  colorimetric assay. Absorbance was quantified by spectrophotometry at λ = 570 nm . IC50 values were calculated by a dose-response analysis using the Prism GraphPad Prism® software.The cells were grown in normal cell culture conditions at 37 °C under a 5% CO


Accurate prediction of anticancer drug responses in cell lines is a crucial step to accomplish the precision medicine in oncology. Although many popular computational models have been proposed towards this non-trivial issue, there is still room for improving the prediction performance by combining multiple types of genome-wide molecular data.We first demonstrated an observation on the CCLE and GDSC datasets, i.e., genetically similar cell lines always exhibit higher response correlations to structurally related drugs. Based on this observation we built a cell line-drug complex network model, named CDCN model. It captures different contributions of all available cell line-drug responses through cell line similarities and drug similarities. We executed anticancer drug response prediction on CCLE and GDSC independently. The result is significantly superior to that of some existing studies. More importantly, our model could predict the response of new drug to new cell line with considerable performance. We also divided all possible cell lines into “sensitive” and “resistant” groups by their response values to a given drug, the prediction accuracy, sensitivity, specificity and goodness of fit are also very promising.CDCN model is a comprehensive tool to predict anticancer drug responses. Compared with existing methods, it is able to provide more satisfactory prediction results with less computational consumption.The online version of this article (10.1186/s12859-019-2608-9) contains supplementary material, which is available to authorized users. The inherent heterogeneity of cancers always makes the same cancer patients exhibiting different anticancer drug responses, which is a major difficulty in cancer treatment. It is critical to accurately predict the therapy responses of patients based on their molecular and clinical profiles , 2. WithBesides the above two types of methods, another important method that gains much attention is the network-based models –19. One Although achieving promising results for certain drugs, most models focused on predicting three types of responses, i.e., ‘old drug to old cell line’, ‘old drug to new cell line’ and ‘new drug to old cell line’ , but paid less attention to the response prediction of ‘new drug to new cell line’. As we all know, updating an existing cancer screen with the latest available drugs and cell lines is not a trivial issue, because it always requires the same expertise, infrastructure and conditions as when the screen was accomplished the first time around. In addition, comprehensive prediction might make potential cancer screen more accurate and experimental design more flexible, as well as accelerate early drug evaluation. Such efforts should be greatly aided by accurate preclinical computational methods.To predict the response of ‘new drug to new cell line’, we should take advantage of all observed (tested or existed) cell line-drug response values. Importantly, two questions need to be asked. The first is whether observed response values have statistical power to predict the response of ‘new drug to new cell line’. The second is how to evaluate the prediction performance of the proposed model. We aim to answer the above two questions.Shivakumar et al. found that structural similarity between drug pairs in the NCI-60 dataset highly correlates with the similarity between their activities across the cancer cell lines . Zhang eCancer Cell Line Encyclopedia (CCLE)  and GenoFor the first question, we want to know whether available drug-cell line response values have the statistical power to predict the response of ‘new drug to new cell line’. Motivated by , 22, we Cell line similarities are measured by Pearson correlation coefficients between their corresponding gene expression profiles. The correlations of most cell line pairs  are larger than 0.8. We divided all possible cell line pairs with correlation coefficients higher than 0.9 into high similar group ‘Hc’, and other pairs into low similar group ‘Lc’.d = 1 − T, where T is the Tanimoto coefficient between drugs Di and Dj. Based on the drug distance matrix  as the Pearson correlation coefficient between cell lines C and Ci, T as the Tanimoto coefficient between drugs D and Dj. Meanwhile, we use R to represent the observed response value of the pair  ∈ Ω. Define Ci and Cj as adjacent if ρ ≠ 0, and the weight of this edge as ρ. Similarly, Di and Dj are called adjacent if their weight T > 0. Define Ci and Dj as adjacent if R is available. Obviously, the resulting network involves cell line similarity and drug similarity information, as well as cell line-drug response situations, so we call it the cell line-drug complex network (CDCN). In fact, this network is the dual-layer integrated cell line-drug network in [We use Ω to represent the set of all possible cell line-drug pairs. Denote twork in . Figure ρ, where the parameter α measures the decay rate with the decrease of ρ. Similarly, define a weight function of drugs τ.Define i, Dj) besides . Based on the generalized observation we are able to make a prediction by dealing with all possible observed response values R as the following,ww reflects the contribution of R to For a given pair , let Ω\{} be the set of all other pairs  and R are not known for any existing drug Dj and any existing cell line Ci). In this circumstance, the cell line-drug response matrix and the corresponding cell line-drug complex network showed in Fig. i, Dj), that isIt is worth mentioning that formula  is appliThe ‘little variation’ is crucial for accomplishing the response prediction of ‘new drug to new cell line’. To highlight the difference between two formulas, we called formula  as CDCN α, τ) could be optimized by minimizing the following overall error functionα and τ are ranged from 0 to 1 with increment 0.01, respectively, and the pair  takes all possible combinations.The decay parameter pairs  and normalized root mean square error (NRMSE) of each drug D were also calculated to assess the model.R are known, and n is the number of such cell lines.Where C ranges over all cell lines for which We executed the following four experiments. (1) Using CDCN model I to predict general responses for the CCLE and GDSC datasets and comparing with six popular computational models. (2) Taking each existed drug-cell line pair as a ‘new drug-new cell line’ pair, we used CDCN model II to predict special responses of these ‘new pairs’, and then compared with the general prediction of model I. (3) Using two models to impute missing data in GDSC independently. (4) Evaluating the model accuracy, sensitivity, specificity and goodness of fit by classifying cell lines into sensitive and resistant groups to some given drug.We first applied CDCN model I to the CCLE dataset with the optimized parameters Next, we conducted CDCN model I for the GDSC dataset with the optimized parameters We used CDCN model II to make a special prediction, i.e. the response prediction of ‘new cell line-new drug’. Fig. We also performed special response prediction for 32 drugs in GDSC with the optimized parameters t0.04,0.1\right $$Scatter plots in Figs. R and R , RDEA119 (fold-change = 2.02 and P = 3.02e-11) and PD-0325901 (fold-change = 1.40 and P = 1.61e-9). Consistently, the predicted response values of wild type cell lines are also higher than that of BRAF mutated cell lines to AZD6244 , RDEA119  and PD-0325901 . In summary, BRAF-mutated cell lines are more sensitive to MEK inhibitors, which is in accordance with the previously published work [The estimation of missing data is considered to be reliable if they exhibit the same or consistent distribution pattern as that by existing data. Following this definition, we first focused on three MEK inhibitors AZD6244, RDEA119, and PD-0325901 in GDSC dataset. Nearly 7% of response values of these three drugs are missing. We found that the predicted missing response values using CDCN models both have a consistent pattern with the existed (observed) response values. We used fold-change and hed work . Similari and Dj is defined as 1- T. We repeated above procedure five times, and used the mean of five Pearson correlation coefficients between predicted and observed response values as the model accuracy. As is shown in Fig. We further compared our method with the nearest neighbor (NN) algorithms in filling up the missing data  to justiWe used a similar method as , 31 to eFor GDSC dataset, our model accuracy is over 60% for 15 of 32 drugs, and over 50% for 27 drugs see Fig. . SensitiHere we should point out that the goodness of fit is relatively small (lower than 0.2) for around half of drugs in both CCLE and GDSC. It is possible even if our model is satisfactory, because CCLE and GDSC are both cross-section datasets, the goodness of fit may be lower because of the variation between the observed values.We further tested our model for Irinotecan in CCLE dataset and Dasatinib in GDSC dataset. As is shown in Fig. There are two key steps for network-based method, i.e., the construction of cell line and drug similarity networks by different types of data and an effective model to execute the prediction. Our method improved the above two steps through an intuitive weighted model which captured different contributions of all available cell line-drug responses. Instead of selecting large plenty of genomic features and making prediction for each drug independently, our model used only two parameters to predict responses for all drugs. This not only decreases the risk of overfitting, but also significantly reduces the computational consumption.As we all know, a main challenge of computational prediction models is how to achieve good performance with low computational consumption. One may take the following efforts to further improve the performance of the model. First, we can integrate other important information, such as copy numbers, gene mutations, drug resistance and transcriptomic signatures of drug sensitivity into the cell line-drug network to get new knowledge. Second, we could further decrease the computational cost by selecting a few informative genes with respect to drug response to construct cell line similarity network instead of using all genes.. Moreover, besides inputting missing values of drug response data, our model could also predict responses of a new drug to existing patients (cell lines), available drugs to a new patient, or even new drugs to new patients. These are more helpful in real clinical practice.We built a simple computational model to comprehensively predict anticancer drug responses. One of the main contributions is to provide a technique to predict the response of “new drug to new cell line”Additional file 1:Table S1. The distance matrix of drugs in CCLE dataset. (XLSX 37 kb)Additional file 2:Table S2. The distance matrix of drugs in GDSC dataset. (CSV 11 kb)Additional file 3:Table S3. Prediction accuracy, sensitivity, specificity and goodness of fit for CCLE dataset. (CSV 6 kb)Additional file 4:Table S4. Prediction accuracy, sensitivity, specificity and goodness of fit for GDSC dataset. (XLSX 34 kb)