Effects of CXCR4 receptor and focal adhesion kinase inhibition in suppressing invasion and overcoming drug resistance in non-small cell lung carcinoma

Rak pluća je najčešće dijagnostikovani tip malignih tumora i vodeći uzrok smrti od malignih oboljenja. Na osnovu kliničke prognoze i odgovora na terapiju čak 85% raka pluća se svrstava u tip nesitnoćelijskog karcinoma pluća (NSCLC). Iako manje agresivan od sitnoćelijskog karcinoma pluća (SCLC), NSCLC lošije odgovara na terapiju. Dva najčešća uzroka neuspeha lečenja NSCLC-a klasičnom hemioterapijom su pojave metastaza i rezistencije na citostatike. Ovi procesi su regulisani brojnim signalnim molekulima, među kojima su i CXC hemokinski receptor 4 (CXCR4) i fokalna adheziona kinaza (FAK), čije ciljano inhibiranje predstavlja obećavajući pristup u lečenju NSCLC-a. Cilj ove studije je bio da se ispita uloga CXCR4 i FAK-a u invazivnosti i rezistenciji NSCLC-a, kao i potencijal inhibicije ovih molekula u supresiji invazije i reverziji rezistencije kod ovog tipa karcinoma pluća. Konkretno, za ispitivanje uloge CXCR4 i FAK-a u invazivnosti NSCLC-a uspostavljen je in vitro sistem ćelija sa različitim funkcionalnim statusom p53 i PTEN tumor supresora, kao i in vivo ortotopni metastatski model NSCLC-a sa p53/PTEN deficijentnim tumorima. U ispitivanom in vitro sistemu istovremena inaktivacija p53 i PTEN tumor supresora dovela je do aktivacije CXCR4 i FAK molekula i nishodnih signalnih molekula AKT-a i ERK-a, što je bilo praćeno povećanjem invazivnog i migratornog potencijala ćelija. Primenom specifičnih inhibitora CXCR4 i FAK-a, WZ811 i PF-573228, pokazan je potencijal primene inhibicije ovih molekula u supresiji invazije i migracije p53-/PTEN- NSCLC ćelija, kao i metastatskog širenja agresivnih p53/PTEN deficijentnih tumora u in vivo modelu. U daljem toku studije za ispitivanje uloge CXCR4 i FAK-a u razvoju rezistencije NSCLC-a na hemioterapiju korišćene su rezistentne ćelijske linije, NCIH460/ R i COR-L23, koje se odlikuju povećanom ekspresijom ABCB1, odnosno ABCC1 transportera. Pokazano je da kombinovani tretmani inhibitorima CXCR4 i FAK-a sa doksorubicinom (DOX) uspešno senzitizuju rezistentne ćelije na pomenuti citostatik. Ispitujući mehanizam reverzije rezistencije utvrđeno je da kombinovani tretmani smanjuju fosforilaciju FAK-a i AKT-a i dovode do aktivacije senescence-a. Istovremeno sa senzitizacijom ćelija na DOX, kombinovanim tretmanima je uspešno suprimirana i invazija rezistentnih ćelija. Imajući u vidu da su rezistencija na terapiju i metataziranje dva najvažnija problema u lečenju NSCLC-a, rezultati studije pokazuju da ciljano inhibiranje CXCR4 i FAK-a, kao i njihova kombinacija sa standardnim hemioterapeuticima, može predstavljati dobar terapeutski pristup u lečenju ovog oboljenja.Lung cancer is the most commonly diagnosed type of malignant tumor and the leading cause of cancer death. Based on clinical prognosis and terapy response, up to 85% of lung cancer is clasified as non-small cell lung carcinoma (NSCLC). Although NSCLC is less agresive than small cell lung carcinoma (SCLC), its response to terapy is worse then SCLC. Two major causes of terapy failure of NSCLC with classic chemotherapeutics are the development of metastasis and chemotherapy resistance. Among other signaling molecules, CXCR4 receptor and focal adhesion kinase (FAK) are known to regulate these processes and their targeted inhibition represents a promising approach in the treatment of NSCLC. The aim of this study was to examine the role of CXCR4 and FAK in the invasiveness and resistance of NSCLC, as well as the potential of their inhibition in the suppression of invasion and the reversal of resistance in this type of lung carcinoma. In order to examine the role of CXCR4 and FAK in the invasiveness of NSCLC, in vitro system of cancer cell lines with different functional status of the p53 and PTEN tumor suppressors was established, as well as in vivo orthoptic metastatic model of NSCLC with p53/PTEN deficient tumors. The simultaneous inactivation of p53 and PTEN tumor suppressors in in vitro system lead to significant increase in invasive and migratory potential associated with activation of CXCR4 and FAK molecules and their downstream signaling molecules, AKT and ERK. The potential for supressing invasion and migration of p53-/PTEN- NSCLC cells, as well as the metastatic spread of aggressive p53/PTEN deficient tumors in in vivo model, has been shown by the application of specific inhibitors of CXCR4 and FAK, PF-573228 and WZ811. Furthermore, resistant cell lines NCI-H460/R and COR-L23 were used to investigate the role of CXCR4 and FAK in the development of NSCLC resistance to chemotherapy. NCI-H460/R and COR-L23 cells are characterized by an increased expression of ABCB1 and ABCC1 transporters, respectively. Our results additionally indicate that inhibition of CXCR4 and FAK in combination with DOX successfully sensitized NSCLC cells to the aforementioned cytostatic. By examining the mechanism of sensitization of NSCLC cells to DOX, it has been discovered that combined treatments reduced phosphorylation of FAK and AKT and lead to activation of senescence. Simultaneously with sensitization of cells to DOX, combined treatment successfully suppressed the invasion of resistant cells. Considering that resistance to therapy and metastasis are the two most important problems in NSCLC treatment, the results of the study show that targeted inhibition of CXCR4 and FAK, as well as their combination with standard chemotherapeutics, could be a promising therapeutic approach in the treatment of this disease

Effects of CXCR4 receptor and focal adhesion kinase inhibition in suppressing invasion and overcoming drug resistance in non-small cell lung carcinoma.

Rak pluća je najčešće dijagnostikovani tip malignih tumora i vodeći uzrok smrti od malignih oboljenja. Na osnovu kliničke prognoze i odgovora na terapiju čak 85% raka pluća se svrstava u tip nesitnoćelijskog karcinoma pluća (NSCLC). Iako manje agresivan od sitnoćelijskog karcinoma pluća (SCLC), NSCLC lošije odgovara na terapiju. Dva najčešća uzroka neuspeha lečenja NSCLC-a klasičnom hemioterapijom su pojave metastaza i rezistencije na citostatike. Ovi procesi su regulisani brojnim signalnim molekulima, među kojima su i CXC hemokinski receptor 4 (CXCR4) i fokalna adheziona kinaza (FAK), čije ciljano inhibiranje predstavlja obećavajući pristup u lečenju NSCLC-a. Cilj ove studije je bio da se ispita uloga CXCR4 i FAK-a u invazivnosti i rezistenciji NSCLC-a, kao i potencijal inhibicije ovih molekula u supresiji invazije i reverziji rezistencije kod ovog tipa karcinoma pluća. Konkretno, za ispitivanje uloge CXCR4 i FAK-a u invazivnosti NSCLC-a uspostavljen je in vitro sistem ćelija sa različitim funkcionalnim statusom p53 i PTEN tumor supresora, kao i in vivo ortotopni metastatski model NSCLC-a sa p53/PTEN deficijentnim tumorima. U ispitivanom in vitro sistemu istovremena inaktivacija p53 i PTEN tumor supresora dovela je do aktivacije CXCR4 i FAK molekula i nishodnih signalnih molekula AKT-a i ERK-a, što je bilo praćeno povećanjem invazivnog i migratornog potencijala ćelija. Primenom specifičnih inhibitora CXCR4 i FAK-a, WZ811 i PF-573228, pokazan je potencijal primene inhibicije ovih molekula u supresiji invazije i migracije p53-/PTEN- NSCLC ćelija, kao i metastatskog širenja agresivnih p53/PTEN deficijentnih tumora u in vivo modelu. U daljem toku studije za ispitivanje uloge CXCR4 i FAK-a u razvoju rezistencije NSCLC-a na hemioterapiju korišćene su rezistentne ćelijske linije, NCIH460/ R i COR-L23, koje se odlikuju povećanom ekspresijom ABCB1, odnosno ABCC1 transportera. Pokazano je da kombinovani tretmani inhibitorima CXCR4 i FAK-a sa doksorubicinom (DOX) uspešno senzitizuju rezistentne ćelije na pomenuti citostatik. Ispitujući mehanizam reverzije rezistencije utvrđeno je da kombinovani tretmani smanjuju fosforilaciju FAK-a i AKT-a i dovode do aktivacije senescence-a. Istovremeno sa senzitizacijom ćelija na DOX, kombinovanim tretmanima je uspešno suprimirana i invazija rezistentnih ćelija...Lung cancer is the most commonly diagnosed type of malignant tumor and the leading cause of cancer death. Based on clinical prognosis and terapy response, up to 85% of lung cancer is clasified as non-small cell lung carcinoma (NSCLC). Although NSCLC is less agresive than small cell lung carcinoma (SCLC), its response to terapy is worse then SCLC. Two major causes of terapy failure of NSCLC with classic chemotherapeutics are the development of metastasis and chemotherapy resistance. Among other signaling molecules, CXCR4 receptor and focal adhesion kinase (FAK) are known to regulate these processes and their targeted inhibition represents a promising approach in the treatment of NSCLC. The aim of this study was to examine the role of CXCR4 and FAK in the invasiveness and resistance of NSCLC, as well as the potential of their inhibition in the suppression of invasion and the reversal of resistance in this type of lung carcinoma. In order to examine the role of CXCR4 and FAK in the invasiveness of NSCLC, in vitro system of cancer cell lines with different functional status of the p53 and PTEN tumor suppressors was established, as well as in vivo orthoptic metastatic model of NSCLC with p53/PTEN deficient tumors. The simultaneous inactivation of p53 and PTEN tumor suppressors in in vitro system lead to significant increase in invasive and migratory potential associated with activation of CXCR4 and FAK molecules and their downstream signaling molecules, AKT and ERK. The potential for supressing invasion and migration of p53-/PTEN- NSCLC cells, as well as the metastatic spread of aggressive p53/PTEN deficient tumors in in vivo model, has been shown by the application of specific inhibitors of CXCR4 and FAK, PF-573228 and WZ811. Furthermore, resistant cell lines NCI-H460/R and COR-L23 were used to investigate the role of CXCR4 and FAK in the development of NSCLC resistance to chemotherapy. NCI-H460/R and COR-L23 cells are characterized by an increased expression of ABCB1 and ABCC1 transporters, respectively. Our results additionally indicate that inhibition of CXCR4 and FAK in combination with DOX successfully sensitized NSCLC cells to the aforementioned cytostatic. By examining the mechanism of sensitization of NSCLC cells to DOX, it has been discovered that combined treatments reduced phosphorylation of FAK and AKT and lead to activation of senescence. Simultaneously with sensitization of cells to DOX, combined treatment successfully suppressed the invasion of resistant cells..

Decreased TSPAN14 Expression Contributes to NSCLC Progression

Tspan14 is a transmembrane protein of the tetraspanin (Tspan) protein family. Different members of the Tspan family can promote or suppress tumor progression. The exact role of Tspan14 in tumor cells is unknown. Earlier, mutational inactivation of the TSPAN14 gene has been proposed to coincide with a low survival rate in NSCLC patients. This study aimed to investigate the correlation of TSPAN14 lack of function with clinicopathological features of NSCLC patients, and to elucidate the role TSPAN14 might have in NSCLC progression. TSPAN14 expression was lower in tumor cells than non-tumor cells in NSCLC patients' samples. The decreased gene expression was correlated with a low survival rate of patients and was more frequent in patients with aggressive, invasive tumor types. Additionally, the role of decreased TSPAN14 expression in the metastatic potential of cancer cells was confirmed in NSCLC cell lines. The highly invasive NSCLC cell line (NCI-H661) had the lowest TSPAN14 gene and protein expression, whereas the NSCLC cell line with the highest TSPAN14 expression (NCI-H460) had no significant metastatic potential. Finally, silencing of TSPAN14 in these non-metastatic cancer cells caused an increased expression of matrix-degrading enzymes MMP-2 and MMP-9, followed by an elevated capacity of cancer cells to degrade gelatin. The results of this study propose TSPAN14 expression as an indicator of NSCLC metastatic potential and progression

Biomimetic tumor engineering to enhance drug discovery - BioengineeredTumor

Development of novel, effective, and safe anti-tumor drugs is still a slow and cumbersome process, which is often attributed to weaknesses of current preclinical assays and low correlation of the preclinical in vitro and in vivo data with the results obtained in clinical trials. Consequently, there is a clear need for development of more reliable in vitro three dimensional (3D) tumor models, which will capture key features of the in vivo tumor cell microenvironment and provide drug testing results relevant for human patients. The aim of the project “Biomimetic tumor engineering to enhance drug discovery – BioengineeredTumor” funded by the Science Fund of the Republic of Serbia is to develop 2 novel, simple and robust 3D models for cultures of carcinoma and osteosarcoma cells by applying systematic and integrated methodology to comprehensively define the key model components. In specific, the aim is to use different human and animal cancer cell lines in conjunction with alginate-based biomaterials as artificial extracellular matrices imitating tumor environments and to cultivate the obtained constructs in perfusion bioreactors providing enhanced transport of nutrients, gases and biochemical signals to the cells as well as adequate levels of hydrodynamic shear stresses. Thus, the strategic goal is to establish an adaptable platform suited to the use by scientists without technical expertise for long-term in vitro studies of cancer cells for applications in anti-cancer drug discovery and validation, development of personalized medical treatments, and cancer research

Anti-invasive effects of CXCR4 and FAK inhibitors in non-small cell lung carcinomas with mutually inactivated p53 and PTEN tumor suppressors

Non-small cell lung carcinoma (NSCLC) is the most common type of lung cancer. At the time of diagnosis, a large percentage of NSCLC patients have already developed metastasis, responsible for extremely high mortality rates. CXCR4 receptor and focal adhesion kinase (FAK) are known to regulate such invasive cancer behavior. Their expression is downregulated by p53 and PTEN tumor suppressors which are commonly co-inactivated in NSCLC patients and contribute to metastasis. Therefore, targeting CXCR4 or FAK seems to be a promising strategy in suppressing metastatic spread of p53/PTEN deficient NSCLCs. In this study, we first examined the invasive characteristics of NSCLC cells with suppressed p53 and PTEN activity using wound healing, gelatin degradation and invasion assays. Further, changes in the expression of CXCR4 and FAK were evaluated by RT-qPCR and Western Blot analysis. Finally, we tested the ability of CXCR4 and FAK inhibitors (WZ811 and PF-573228, respectively) to suppress the migratory and invasive potential of p53/PTEN deficient NSCLC cells, in vitro and in vivo using metastatic models of human NSCLC. Our results showed that cells with mutually inactive p53 and PTEN have significantly increased invasive potential associated with hyperactivation of CXCR4 and FAK signaling pathways. Treatments with WZ811 and PF-573228 inhibitors significantly reduced migratory and invasive capacity in vitro and showed a trend of improved survival in vivo. Accordingly, we demonstrated that p53/PTEN deficient NSCLCs have extremely invasive phenotype and provided a rationale for the use of CXCR4 or FAK inhibitors for the suppression of NSCLC dissemination.This is a post-peer-review, pre-copyedit version of an article published in Investigational New Drugs. The final authenticated version is available online at: [http://dx.doi.org/10.1007/s10637-017-0494-4

Association of Overexpressed MYC Gene with Altered PHACTR3 and E2F4 Genes Contributes to Non-Small Cell Lung Carcinoma Pathogenesis

Background: C-Myc is one of the major cellular oncogenes overexpressed in non-small cell lung carcinoma (NSCLC). Its deregulated expression is necessary but not sufficient for malignant transformation. We evaluated expression of MYC gene in NSCLC patients and its association with alterations in the genes previously identified to be related to NSCLC pathogenesis, PHACTR3 and E2F4. Methods: We analyzed MYC gene expression by qRT-PCR in 30 NSCLC patients' samples and paired normal lung tissue. MYC expression was further statistically evaluated in relation to histopathological parameters, PHACTR3 and E2F4 gene alterations and survival. Alterations in aforementioned genes were previously detected and identified based on AP-PCR profiles of paired normal and tumor DNA samples, selection of DNA bands with altered mobility in tumor samples and their characterization by the reamplification, cloning and sequencing. Results: MYC expression was significantly increased in NSCLC samples and its overexpression significantly associated with squamous cell carcinoma subtype. Most importantly, MYC overexpression significantly coincided with mutations in PHACTR3 and E2F4 genes, in group of all patients and in squamous cell carcinoma subtype. Moreover, patients with jointly overexpressed MYC and altered PHACTR3 or E2F4 showed trend of shorter survival. Conclusions: Overall, MYC is frequently overexpressed in NSCLC and it is associated with mutated PHACTR3 gene, as well as mutated E2F4 gene. These joint gene alterations could be considered as potential molecular markers of NSCLC and its specific subtypes

Инхибиција аутофагије сензитизује ћелије глиобластома на инхибиторе Срц тирозин-киназе, деривате пиразоло[3,4- д]пиримидина Si306 и pro-Si306

Glioblastom je jedan od najagresivnijih tumora mozga koji karakteriše infiltrirajuća priroda, intenzivna proliferacija i rezistencija na terapiju. Ćelije glioblastoma imaju visoku ekspresiju Src tirozin-kinaze koja reguliše proliferaciju, preživljavanje i invazivnost tumorskih ćelija čineći je potencijalnom metom za terapiju. Inhibitori tirozin-kinaza mogu indukovati autofagiju koja deluje protektivno na tumorske ćelije. Sposobnost inhibitora Src tirozin-kinaze, derivata pirazolo[3,4-d]pirimidina Si306 i njegovog proleka pro-Si306, da indukuju autofagiju ispitana je na ćelijskoj liniji humanog glioblastoma U87 i njenoj varijanti sa višestrukom rezistencijom na lekove U87-TxR. Tretman ovim jedinjenjima uzrokovao je pojavu autofagozoma u ćelijama nakon 3 sata, a efekat na indukciju autofagije opstao je i nakon 48 sati što je utvrđeno analizom markera autofagije LC3 i p62. Inhibicija autofagnog fluksa bafilomicinom A1 značajno je uvećala postojeće anti-proliferativno dejstvo Si306 i pro-Si306. Takođe, kombinovani tretmani Src inhibitora sa bafilomicinom A1 doveli su do nekroze nakon 48 sati. Dobijeni rezultati sugerišu da autofagija indukovana ovim jedinjenjima ima zaštitnu ulogu u ćelijama glioblastoma i da se modulacija autofagije može koristiti za senzitizaciju ćelija glioblastoma na inhibitore Src tirozin-kinaze. Pored toga, pomenuti efekti Si306 i pro-Si306 nisu umanjeni prisustvom višestruko- rezistentnog fenotipa, što ovim jedinjenjima daje potencijal za lečenje rezistentnih tumora.Глиобластом је један од најагресивнијих тумора мозга који карактерише инфилтрирајућа природа, интензивна пролиферација и резистенција на терапију. Ћелије глиобластома имају високу експресију Срц тирозин-киназе која регулише пролиферацију, преживљавање и инвазивност туморских ћелија чинећи је потенцијалном метом за терапију. Инхибитори тирозин-киназа могу индуковати аутофагију која делује протективно на туморске ћелије. Способност инхибитора Срц тирозин-киназе, деривата пиразоло[3,4-д]пиримидина Si306 и његовог пролека pro-Si306, да индукују аутофагију испитана је на ћелијској линији хуманог глиобластома U87 и њеној варијанти са вишеструком резистенцијом на лекове U87-TxR. Третман овим једињењима узроковао је појаву аутофагозома у ћелијама након 3 сата, а ефекат на индукцију аутофагије опстао је и након 48 сати што је утврђено анализом маркера аутофагије LC3 и p62. Инхибиција аутофагног флукса бафиломицином А1 значајно је увећала постојеће анти-пролиферативно дејство Si306 и pro-Si306. Такође, комбиновани третмани Срц инхибитора са бафиломицином А1 довели су до некрозе након 48 сати. Добијени резултати сугеришу да аутофагија индукована овим једињењима има заштитну улогу у ћелијама глиобластома и да се модулација аутофагије може користити за сензитизацију ћелија глиобластома на инхибиторе Срц тирозин-киназе. Поред тога, поменути ефекти Si306 и pro-Si306 нису умањени присуством вишеструко- резистентног фенотипа, што овим једињењима даје потенцијал за лечење резистентних тумора.Knjiga sažetaka: Treći Kongres biologa Srbije, Zlatibor, Srbija 21 - 25. 9. 2022

On optimal temozolomide scheduling for slowly growing glioblastomas

Background: Temozolomide (TMZ) is an oral alkylating agent active against gliomas with a favorable toxicity profile. It is part of the standard of care in the management of glioblastoma (GBM), and is commonly used in low-grade gliomas (LGG). In-silico mathematical models can potentially be used to personalize treatments and to accelerate the discovery of optimal drug delivery schemes. Methods: Agent-based mathematical models fed with either mouse or patient data were developed for the in-silico studies. The experimental test beds used to confirm the results were: mouse glioma models obtained by retroviral expression of EGFR-wt/EGFR-vIII in primary progenitors from p16/p19 ko mice and grown in-vitro and in-vivo in orthotopic allografts, and human GBM U251 cells immobilized in alginate microfibers. The patient data used to parametrize the model were obtained from the TCGA/TCIA databases and the TOG clinical study. Results: Slow-growth "virtual" murine GBMs benefited from increasing TMZ dose separation in-silico. In line with the simulation results, improved survival, reduced toxicity, lower expression of resistance factors, and reduction of the tumor mesenchymal component were observed in experimental models subject to long-cycle treatment, particularly in slowly growing tumors. Tissue analysis after long-cycle TMZ treatments revealed epigenetically driven changes in tumor phenotype, which could explain the reduction in GBM growth speed. In-silico trials provided support for implementation methods in human patients. Conclusions: In-silico simulations, in-vitro and in-vivo studies show that TMZ administration schedules with increased time between doses may reduce toxicity, delay the appearance of resistances and lead to survival benefits mediated by changes in the tumor phenotype in slowly-growing GBMs.This research was funded by the James S. Mc. Donnell Foundation (USA) 21st Century Science Initiative in Mathematical and Complex Systems Approaches for Brain Cancer (Collaborative award 220020560, doi:10.37717/220020560); Ministry of Education, Science and Technological Development, Republic of Serbia (ref. number 451-03-9/2021-14/200007); Ministerio de Ciencia e Innovación and FEDER funds, Spain (grant number PID2019-110895RB-I00, doi: 10.13039/501100011033 to VMP-G, and RTI2018-093596 and PI21CIII/00002 to PS-G); and Universidad de Castilla-La Mancha (grant number 2020-PREDUCLM-15634 to JJ-S).S

New anti-glioblastoma strategy with natural compounds sclareol and doxorubicin

Background: Doxorubicin (DOX) has been very effective against glioblastoma invitro. Its application in vivo is hampered because it cannot pass the blood–brainbarrier (BBB). Significant research efforts are invested to overcome this limitation.Sclareol (SC) is an aromatic compound naturally found in clary sage. Thecombination of SC and DOX showed promising effects in different tumor types invitro and in vivo. Therefore, we tested their combination and innovative hybridmolecules (SC:DOX) on glioblastoma cells with the expression of P-glycoprotein, amajor component of BBB and cancer multidrug resistance marker. Methods:Cytotoxicity and selectivity towards glioblastoma cells of SC, DOX, theircombination, and SC:DOX were examined by MTT assay. The effect of SC on DOXaccumulation was determined by flow cytometry. We also studied SC:DOXaccumulation, cellular uptake, localization imaging, and DNA damage induction.Results: The effects of simultaneous SC and DOX treatments demonstrated theconsiderable potential of SC to reverse DOX resistance in glioblastoma cells andincrease DOX accumulation. SC:DOX hybrids, named CON1 and CON2 were lesscytotoxic than DOX, but with reduced resistance and increased selectivity towardsglioblastoma cells. Cellular uptake of CON1 and CON2 was increased in glioblastomacells compared to DOX. Perinuclear localization of CON1 and CON2 vs. nuclearlocalization of DOX as well as no DNA damaging effects suggest a differentmechanism of action for SC:DOX. Conclusion: The combination of SC and DOX, andtheir innovative hybrids, could be considered a promising strategy that can overcomethe limitations of DOX application in glioblastoma.Kanazir S, Savić D, editors. Book of abstracts: 8th Congress of Serbian neuroscience society with international participation; 2023 May 31 - Jun 2; Belgrade, Serbia. Belgrade : Serbian Neuroscience Society; 2023. p. 71

Adaptable alginate-based microfibers for 3D in vitro cultures of cancer cells: an anticancer drug testing model

The slow advance in anticancer drug development can be attributed to the limitations of conventional models, predominantly monolayer cell (2D) cultures and animal models, which inadequately recapitulate the complex nature of human malignant tumors. Three-dimensional (3D) in vitro models are invaluable tools in drug screening; however, creating a universal model for all cancer types poses challenges due to the diverse nature of cancers. The aim of this work was to develop a single, versatile model using alginate microfibers to accommodate cultivation of various cancer cells