Procter & Gamble (United Kingdom)
Digital Repository of Archived Publications - Institute for Biological Research Sinisa Stankovic (RADaR)Not a member yet
7316 research outputs found
Sort by
Co-occurrence networks of chironomid species (Diptera, Chironomidae) as a measure of ecosystem stability
No full textUnderstanding ecological stability is crucial for large river ecosystems exposed to multipleanthropogenic stressors. Chironomids are important benthic organisms, but their complex assemblagedynamics, especially their persistence and variability, require deeper analysis and understanding. Thisstudy aimed to assess 1) the spatio-temporal variation of chironomid diversity across three Danubesectors over three Joint Danube Surveys expeditions; 2) analyse potential species associations usingco-occurrence networks and 3) evaluate assemblage stability by quantifying persistence and variabilityof network structure over time. We identified 169 chironomid taxa in the Danube, with taxa richnessdecreasing downstream. Linear discriminant analysis confirmed distinct assemblages per sector. Cooccurrence networks for each sector and year were compared using differential network analysis.Temporal comparisons revealed striking lack of persistence: year-specific (gamma) links dominatednetwork changes, while few stable, persistent (alpha) links were found between taxa across six-yearintervals. Sector 1 (Upper Danube) showed lower network connectivity compared to Sectors 2 (MiddleDanube) and 3 (Low Danube), potentially reflecting different environmental pressures. The hightemporal turnover in network structure suggests Danube chironomid assemblages are highly dynamic,possibly indicating resilience through compositional changes, rather than static persistence. Analysis of co-occurrence networks reveals community dynamics, thus complementing traditional diversity metricsand holding potential for environmental monitoring in large rivers
Comprehensive Evaluation of the Flavonoid Profile of Fourteen Nepeta Species Using HPLC-HRMS Technique
No full textThe aim of this research was to examine the detailed flavonoid profile in the leaves of fourteen phyllodiverse Nepeta taxa (N. rtanjensis, N. grandiflora, N. laevigata, N. sibirica, N. cataria, N. racemosa, N. parnassica, N. nuda, N. nervosa, N. stewartiana, N. ernesti-mayeri, N. clarkei, N. subsessilis, and N. govaniana) grown under greenhouse conditions. Using LC/MS technique, 60 flavonoids were identified, of which there were 3 flavonoid C-glycosides, 42 flavonoid O-glycosides, and 12 flavonoid aglycones. Relative quantification data (peak areas) were subjected to statistical analysis and visualized using hierarchical cluster analysis (HCA) to adequately interpret the interrelations between samples. Most of the identified flavonoid derivatives belonged to the flavone subgroup (angularly methylated apigenin and luteolin), which was expected. Surprisingly, some compounds were species-specific. Thus, apigenin 6,8-C-di-hexoside, naringenin 7-O-hexoside and pectolinarigenin 7-O-hexuronide were found only in samples of N. nervosa, while quercetin 3-O-(2"-caffeoyl-6"-rhamosyl)-hexoside-7-O-hexuronide was found only in samples of N. ernesti-mayeri. Kaempferol 3-O-[6"-(3-hydroxy-3-methylglutaroyl)]-hexoside and chrysoeriol 7-O-[2"-(5'''-acetyl)-pentoyl]-pentoside were detected only in N. govaniana, while luteolin 3′,4′-dimethyl ether 7-O-(6"-rhamnosyl)-hexoside and chrysoeriol 7-O-(6'''-rhamnosyl)-acetyl-hexoside were identified only in samples of N. clarkey. N. laevigata was the only one of the examined species in which low content of flavonoids was detected, with only two recorded compounds, apigenin 7-O-(2"-hexuronyl)-hexuronide and chrysoeriol 7-O-hexoside, present in trace amounts. Since certain flavonoids were only recorded in some of the analyzed taxa, this study may be the first attempt to propose species-specific biomarkers.Book of abstracts: International Conference on Natural Sciences and Biotechnology: Kliment’s Days 2025: with satellite scientific conference: Reintroduction of Conservation: Reliant species; 2025 Nov 6-7; Sofia, Bulgaria. Sofia: The Faculty of Biology at Sofia University “St. Kliment Ohridski”; 2025. p. 189
Antioxidant Supplementation with ProCloSupp Protects Against Renal Toxicity of Atypical Antipsychotics in Rats: Implications for Safer Treatment Strategies
No full textAtypical antipsychotics (AAP), including clozapine (Clo), aripiprazole (Ari), and risperidone
(Ris), are widely used in psychiatry but can lead to kidney damage due to oxidative
stress. This study investigated whether dietary supplementation with selected
antioxidants—ellagic acid, vitamin C, zinc, and seleno-methionine (SeMet) in fish oil,
formulated as the composite product “ProCloSupp” (PCS)—can mitigate the oxidative
damage induced by subchronic administration of AAP. Rats were treated with each antipsychotic
for 28 days, with PCS added in the last 14 days. The kidney tissue was examined
histologically and by determining the activities of antioxidant enzymes (copper, zinc and
manganese superoxide dismutase—CuZn SOD and Mn SOD, catalase—CAT, glutathione
peroxidase—GPx, glutathione reductase—GR, glutathione S-transferase—GST). All AAPs
caused discrete to moderate renal damage and significant changes in enzyme profiles,
which were most pronounced with Ari. Clo and Ari significantly decreased CuZn SOD and
Mn SOD activity, while Ris only affected Mn SOD. Clo additionally increased CAT activity,
while Ari increased GPx activity. Antioxidant-related protein levels increased only in the
Ris group. PCS supplementation increased CuZn SOD and GPx activities and was associated
with less pronounced histopathological changes than antipsychotic treatment alone. In
conclusion, subchronic Clo, Ari, and Ris exposure induces oxidative renal damage in rats,
while PCS supplementation enhances antioxidant defences and attenuates tissue damage.
These results support PCS as a potential nephroprotective strategy in antipsychotic therapy
Agmatine and the inflamed microglia: a preclinical pathway to neuroprotective strategies
No full textChronic neuroinflammation is a hallmark of many neurodegenerative diseases, making microglia cultures an indispensable tool in preclinical research and therapy discovery. In this study, we used an in vitro model of neuroinflammation, murine BV-2 microglial cells stimulated with lipopolysaccharide (LPS), to investigate the therapeutic potential of agmatine, an endogenous polyamine, in modulating inflammation-induced metabolic and oxidative dysregulation. Through integrated analyzes of gene and protein expression (qPCR, Western blot, ELISA), mitochondrial function (fluorescent probes for membrane potential and
superoxide) and cellular bioenergetics (lactate release, ATP levels), we demonstrated that LPS induces a shift toward glycolysis in microglia via the PI3K/Akt/mTOR/HIF-1α axis, leading to mitochondrial dysfunction, oxidative stress and proinflammatory cytokine production. Agmatine pretreatment mitigated these effects by downregulating mTOR/HIF-1α signaling, lowering lactate and TNF levels, and restoring mitochondrial function. In addition, agmatine increased intracellular ATP even under inflammatory condition, suggesting a shift toward energy-efficient metabolic reprogramming. In parallel, agmatine
attenuated LPS- induced nitrosative and oxidative stress by decreasing iNOS and xanthine oxidase activity, suppressing NF-κB signaling, and promoting antioxidant defense through Nrf2 activation, glutathione upregulation, and increased activity of GPx, GR, and CAT. This dual modulation of metabolic and redox pathways promoted a phenotypic shift of microglia toward an antiinflammatory state characterized by increased ARG1, CD206, and HO-1 expression and decreased release of TNF, IL-1 beta,
and IL-6. Interestingly, agmatine alone also induced mild oxidative stress and triggered adaptive antioxidant responses, suggesting a potential priming mechanism that increases cellular resilience to subsequent inflammatory insults. Overall, our results emphasize the utility of microglia models in preclinical research and position agmatine as a promising candidate for therapeutic modulation of neuroinflammation.Abstract Book: Nature Conferences: Engineered Models of Human Disease; 2025 Oct 28-29; Belgrade, Serbia. Berlin: Springer Nature; 2025
From tumors to organoids: establishing an advanced colorectal carcinoma model to uncover molecular drivers of compensatory proliferation
No full textCompensatory proliferation refers to the enhanced division of progenitor cells following tissue damage or loss, restoring structure and function. In advanced cancer, this process powerfully promotes tumor growth, resilience, and relapse after therapy. To test new strategies, an “alternative” mouse model of advanced colorectal carcinoma (CRC) was developed by inoculating a mixture of live and dead MC38 cells, simulating close communication of dying and viable cell fractions in the tumor microenvironment. The goal was to compare transcriptional profiles between the “alternative” and “classically” induced tumor models with corresponding organoids, to assess how they faithfully represent tumors and their suitability for drug testing. RNA sequencing identified 4,548 genes with significantly differential expression, with 1,185 upregulated in organoid-derived and 3,363 in tumor-derived samples. Tumors showed enrichment in immune-related pathways (immune processes and regulation, cell adhesion, responses to external stimuli), whereas organoids displayed changes in a metabolic signature (sugar metabolism, hypoxia response, redox regulation, detoxification of drugs). In addition, RNA sequencing discovered five protein-coding and long noncoding genes, reflecting differences in organoids derived from “classical” (OL) and “alternative” (OLD) models, thus highlighting the successful establishment of an advanced CRC model. While Ptprtos (lncRNA) and Clec4d illustrated intensive abolishment of tumor suppressor activities and establishment of immunosuppressive ambient in OL, respectively, dominance of lncRNA-like E13030I04Rix, Brdt, and Gpm6d genes in OLD indicated potentiation of stemness, plasticity, chronic inflammation, and regeneration as hallmarks of novel organoids. Overall, despite lacking full immune complexity, organoids preserve tumor-associated metabolic states, making them valuable for preclinical drug testing. Most importantly, the comparative analysis of OL and OLD highlights the translational potential of the “alternative” model for identifying novel diagnostic markers in CRC and therapeutic strategies that multiply affect the CRC microenvironment.Abstract Book: Nature Conferences: Engineered Models of Human Disease; 2025 Oct 28-29; Belgrade, Serbia. Berlin: Springer Nature; 2025
Dark side of extraction of lithium, boron and arsenic: impact on local amphibans and reptiles
No full textDue to energy crisis, European governements showed increased interest in continentwide exploitation of lithium as a valuable source. In the process of lithium extraction from the ground, another two chemical elements are released to the environment: boron and arsenic. Being substantially concerned about impacts of such mining projects on local biodiversity, we surveyed literature about the known impact of lithium, boron and arsenic on amphibians and reptiles. We found surprisingly small amount of information (21 and 10 references in total for amphibians and reptiles, respectively). However, these scarce literature data are revealing somewhat disturbing facts: Exposure of amphibians to lithium (10 references found in total), boron (two references in total) or arsenic (9 references found in total), especially in aquatic ecosystems, lead to a wide range of adverse effects, both for individuals at different developmental stages and for whole populations. In some of the species studied, reduced survival of juveniles and reduced fecundity of adults were observed. Current knowledge on the impact of lithium on reptile populations indicates that the release of lithium from soil into the environment leads to the disruption of local populations, and, in the long term, possibly to their extinction (one reference found in total). In snakes
and lizards, a negative effect of arsenic accumulation has been observed in phenotypic traits essential for survival; moreover, arsenic accumulated in the bodies of females is transferred to their offspring (9 references found in total). No studies about impact of boron on reptiles were found. In conclusion, the effects of lithium, boron and arsenic extraction from the ground on local amphibian and reptile populations require more thorough research.Abstract booklet: Oral & Poster presentations: 23rd European Congress of Herpetology: SEH2025; 2025 Sep 8-12; Bonn, Germany. Societas Europaea Herpetologica; 2025. p. 41
Synergistic Effects of Genipin and Alendronate in 3D-Bioprinted Gelatin-Polyvinylpyrrolidone Scaffolds
No full textIn this study, we introduce a one-step semi-solid extrusion 3D printing strategy to fabricate gelatin-polyvinylpyrrolidone
(GAG-PVP) scaffolds loaded with a low dose (0.5 wt%) of alendronate (ALN) and crosslinked in situ with 1 wt% genipin.
The genipin-crosslinked ALN scaffold (GAG-PVP-GEN-ALN) demonstrated enhanced functional performance compared
to both non-crosslinked GAG-PVP and GAG-PVP-ALN controls. Its peak swelling reached 462% at 5 h, surpassing the
362% of the unmodified scaffold and preventing the rapid dissolution observed for GAG-PVP-ALN, before gradually
deswelling for 96 h. Water contact angle measurements confirmed that genipin fully restored surface hydrophobicity
(101.9°), counteracting the pronounced wettability induced by ALN (47.8°) and exceeding the 78.2° of the GAG-PVP
matrix, which is consistent with swelling ratio. Differential scanning calorimetry (DSC) indicated enhanced thermal stability of the crosslinked gelatin, with shifts in both glass transition and denaturation temperatures reflecting greater molecular
rigidity despite the presence of glycerol as a plasticizer. Mechanical testing showed that while alendronate alone reduced
mechanical performance, the combined inclusion of alendronate and genipin significantly enhanced scaffold properties
compared to gelatin-polyvinyl pyrrolidone blend: tensile strength increased from 19.7 MPa to 39.8 MPa, elastic modulus rose from 805 MPa to 1174 MPa, and microhardness improved from 9.24 MPa to 22.3 MPa, values nearing those
of native cancellous bone. The sustained ALN release profile extended from an abrupt 3 h burst in GAG-PVP-ALN to
a controlled 48 h delivery in GAG-PVP-GEN-ALN, following first-order kinetics. Both direct and indirect cytotoxicity
assays confirmed high cell viability (>85%) without morphological abnormalities. These results highlight that embedding
low-dose ALN within a genipin-crosslinked gelatin-PVP network results in a mechanically robust, biocompatible scaffold with tunable swelling and prolonged drug release, offering a versatile platform for localized bone tissue engineering
Nrf2-driven redox coupling of tumour and associated adipose tissue during early tumour growth in an orthotopic breast cancer model
No full textThe redox-based two-way communication between breast cancer cells and their tumour microenvironment (TME) contributes significantly to the establishment of the malignant phenotype. Due to its well-established role in redox-metabolic reprogramming, Nuclear Factor Erythroid 2–Related Factor 2 (Nrf2) has emerged as a potentially key player in the bilateral interaction between breast cancer cells and cancer-associated adipose tissue (CAAT), the main cellular component of the TME. In this study, we used an orthotopic model of Nrf2+/+ luminal type B breast cancer in wild-type (WT) mice and mice lacking functional Nrf2 (Nrf2KO) to investigate the role of Nrf2-driven redox coupling between breast cancer and CAAT. To this end, we examined the expression profiles, localisation, and activity of the main antioxidant defence (AD) enzymes in breast tumour tissue in the Nrf2+/+ or Nrf2−/− host environment at different phases of early tumour growth. In addition, we analysed key enzymes involved in nicotinamide adenine dinucleotide phosphate (NADPH) metabolism. We demonstrate the establishment of a distinct redox profile of breast cancer cells in the Nrf2−/− TME when compared to WT TME. Furthermore, the activity and protein levels of AD enzymes in CAAT showed both Nrf2-dependent and/or tumour-dependent changes. Taken together, these results highlight the importance of Nrf2-driven modulations in the host TME for establishing the redox profile of breast cancer. Moreover, the initial phase of early tumour growth appears most susceptible to the absence of functional Nrf2 in the host TME, thus providing a new potential therapeutic target point for breast cancer therapy
In Vitro Shoot Cultures of Micromeria graeca: Micropropagation and Evaluation of Methanolic Extracts for Anticancer and Antimicrobial Activity
No full textMicromeria graeca (L.) Benth. ex Rchb. (Lamiaceae) is a promising medicinal plant valued for its antioxidant, anti-hyperglycemic, anti-hypertensive, antimicrobial, and anti-aflatoxigenic properties. It is rich in phenolic and flavonoid compounds, supporting its traditional use for digestive, respiratory, cardiovascular, and dermatological conditions. Plant tis-sue culture facilitates controlled in vitro propagation to study plant growth and bioactive properties. The effects of activated charcoal and varying subculture intervals on multiplication and biomass production in M. graeca shoot cultures were investigated. The phenolic composition of methanolic extracts from in vitro-grown plants was characterized using high-performance liquid chromatography (HPLC), identifying rosmarinic, caffeic, and syringic acids as the primary phenolic compounds. Antimicrobial activity against selected microbial strains was evaluated using a micro-well dilution assay. Anticancer activity of selected extracts was assessed in human hepatocellular carcinoma cell line HepG2, with flow cytometry (Annexin-V/PI staining) used to analyze cell death mechanisms, and com-pared to pure rosmarinic acid (RA). Activated charcoal showed no beneficial effects on multiplication or biomass production, but significantly increased phenolic acid content (up to 4-fold). RA dominated the phenolic profiles, with other phenolic acids present in lower amounts. Methanolic extracts exhibited negligible antimicrobial activity compared to reference antibiotics and fungicide. Extracts from 4-week-old shoot cultures displayed modest anti-hepatoma activity (IC50 values of CV assay ranging from 193 to 274 µg mL−1), inducing HepG2 cell apoptosis via oxidative stress, independent of RA. Our results suggest that the metabolic output of M. graeca shoot cultures and consequently their biological activity can be modulated by varying in vitro culture conditions. These findings underscore the potential of their methanolic extracts for biotechnological production and therapeutic applications
Personalized Genomic Analysis for Clinical Insights in Non-Small Cell Lung Cancer
No full textBackground: Non-small cell lung carcinoma (NSCLC) accounts for approximately 85% of all lung cancer cases and
remains the leading cause of cancer-related mortality worldwide [1]. Clinical treatment has improved significantly in
recent years with the introduction of targeted therapies and immune checkpoint inhibitors. Therapeutic decisions are
largely guided by the identification of somatic genetic driver alterations in tumors [2]. However, this approach alone is
often insufficient to capture the complexity of tumor biology and functional response to treatment.
Decreasing sequencing costs and advances in the analysis of sequencing data have opened up the possibility of using
whole-exome sequencing (WES) in the clinic. Applying this approach to genomic profiling enables the identification
of all potential driver mutations, tumor mutational burden, homologous recombination deficiency, mismatch repair
deficiency, copy number alterations, mutational signatures and even germline information for each patient [3]. By
overcoming the limitations of targeted panels, WES facilitates more personalized characterization of tumor biology,
which is important for optimizing precision oncology approaches [4,5]. In parallel, functional testing of the patientderived tumor cells in response to drugs offers a direct way of assessing treatment sensitivity [6]. Drug testing on
primary cell cultures, including chemotherapeutics and tyrosine kinase inhibitors (TKIs), enables real-time assessment
of cytotoxicity and treatment sensitivity that can be directly correlated with the underlying genomic landscape [7].
By integrating functional drug response data with WES-based genomic profiling, our approach aims to provide a
potentially comprehensive diagnostic platform for NSCLC. This combined methodology not only identifies clinically
actionable genomic features, but also validates their functional relevance, thus improving predictive power. It also allows
us to identify rare, exceptional responders in unselected patients, which is not possible with current patient selection
strategies. Ultimately, this integrative approach represents a step towards truly personalized cancer diagnostics, where
both the molecular and functional characteristics of the tumor are taken into account in clinical decision-making.
Methodology: Collection of NSCLC tissue samples. Tissue samples from 67 NSCLC patients were collected from the
Clinic for Thoracic Surgery at the University Clinical Center of Serbia with informed consent obtained from the patients
and approved by the Ethics Committee of the University Clinical Center of Serbia (approval reference number 623/4).
Samples were collected during surgery and histopathologically examined to confirm the diagnosis of NSCLC, histologic
grade, stage, necrosis and status of lymph node invasion. Fresh tumor tissue destined for primary cell culture was
placed in sterile tubes containing an antibiotic–antimycotic solution and immediately transported to the research
laboratory for further processing. For WES, paired tumor and corresponding normal tissue samples were frozen in
liquid nitrogen immediately after surgical removal and stored in liquid nitrogen until DNA isolation.
Patent-derived NSCLC cell cultures. Patient-derived NSCLC cell cultures containing both cancer cells and stromal
components were established from freshly collected NSCLC tumor samples. These primary cultures were maintained in vitro for 1 – 2 weeks to ensure cell attachment, growth and stabilization prior to drug testing.
Fluorescence immunoassay. An immunofluorescence assay was developed to screen patient-derived NSCLC cell cultures
for their response to chemotherapeutics and TKIs. Drug concentrations were selected based on clinically relevant
exposure levels, with the maximum plasma concentration (Cmax) as the upper limit and four lower concentrations to
capture dose-dependent effects. Automated microscopy (ImageXpress Pico, Molecular Devices, San Jose, CA, USA) and
image analysis with CellReporterXpress software were used to quantify total cell number and differentiate cancer cells
from stromal cells based on cytokeratin (CK8/18) expression. The cytotoxicity was determined using the Cell Scoring
Analysis Protocol, which evaluates changes in the viability of cancer cells under different treatment conditions.
WES. Genomic DNA was extracted from paired tumor and corresponding normal tissue samples from NSCLC patients
using the QIAamp Fast DNA Tissue Kit. Sequencing was performed by Novogene Co (Cambridge, UK). Library preparation
was performed using the SureSelect Human All Exon V6 Capture Kit, and paired-end 150 bp reads were generated on
the Illumina NovaSeq 6000 platform. At Novogene, raw sequencing reads were subjected to initial quality control
using fastp (version 0.23.1), clean reads were aligned to the human reference genome (GRCh37) using the BWA-MEM
algorithm (version 0.7.8-r455), and post-alignment processing was performed with the Picard toolkit (version 2.6.0).
The processed BAM files containing high quality aligned reads were provided to our team. Subsequent bioinformatic
analyses were performed in-house using a pipeline developed in R/Bioconductor and shell-based tools. Somatic single
nucleotide variants (SNVs) and small insertions/deletions (InDels) were identified using VarScan2 (version 2.4.3), with
tumor normal pair analysis allowing accurate discrimination between somatic events and germline polymorphisms.
For germline variant detection, the GATK HaplotypeCaller (version 4.2) was used to generate high-confidence germline
variant call sets. Structural variants (SVs) were detected using Manta, which identifies genomic rearrangements such
as translocations, inversions and large insertions/deletions. Copy number variations (CNVs) were profiled using CNVkit,
which provides genome-wide CNV profiles at exome resolution. PureCN was used to determine tumor purity and ploidy
as well as allele-specific copy number states. Variant annotation was performed with the Variant Effect Predictor (VEP)
(ensembl-vep-release-105), which integrates several annotation resources, including RefSeq, dbSNP, COSMIC, ClinVar,
1000 Genomes, NHLBI-ESP, gnomAD, SIFT, PolyPhen, and HGMD-PUBLIC. This annotation workflow provided detailed
information on the functional consequences of each variant, pathogenicity predictions and clinical relevance. Analysis
of mutational signatures was performed using the R package signature.tools.lib, which decomposes the observed
mutational spectra into known COSMIC signatures for single base substitutions (SBS), double base substitutions
(DBS) and indel signatures (ID) and detects potential novel mutational processes active in the tumors. All results were
integrated, curated and analyzed using R (v4.3) and Bioconductor. Somatic and germline variants, CNV profiles and
mutational signatures were summarized with custom scripts and visualized with tools such as maftools and circlize. The
Cell Scoring Analysis Protocol described above was combined with the WES results to identify potential correlations
between genetic alterations and drug response.
Results: To investigate the therapeutic response of NSCLC patient cells, we performed ex vivo drug testing with a
range of chemotherapeutic agents and TKIs currently used in clinical practice. The cell response of NSCLC patients was
assessed using a modified scoring system previously described [8], which integrates multiple weighted parameters
to provide an overall drug response score. This approach allowed quantitative comparison of drug effects on cancer
and stromal cells as assessed by immunofluorescence-based cell classification. The analysis revealed considerable
variability in sensitivity profiles between patients, with some drugs showing strong cytotoxic effects in certain cultures
while being ineffective in others.
To complement the ex vivo drug sensitivity testing and gain deeper insight into the molecular features that influence
treatment response, we performed WES of tumor and corresponding normal tissue in each NSCLC patient. This
genomic profiling allowed us to identify somatic mutations, germline variants, copy number alterations and mutational
signatures that may underlie the differences in therapeutic sensitivity observed in the patient-derived cultures. The
mutational landscape of individual patients varied widely, reflecting the heterogeneity of NSCLC and underscoring the
need for individualized therapeutic approaches.
The value of this approach is demonstrated in patients TR33 and TR106. In patient TR33, a smoker, whole-exome
sequencing revealed a high number of somatic SNVs. Mutational signature analysis revealed a dominant presence of
the COSMIC signature SBS4, which is associated with tobacco exposure, while no clinically actionable point mutations
were detected. However, copy number analysis revealed amplification of the HER2 gene. Despite the absence of
actionable mutations, in ex vivo assays this patient’s cells responded selectively and robustly to the EGFR inhibitor
erlotinib, while remaining largely resistant to standard chemotherapeutics. In contrast, patient TR106 had a lower total
number of somatic SNVs but had amplification of the PIK3CA gene and a PTEN missense mutation accompanied by loss
of heterozygosity. Mutational signature analysis revealed the presence of SBS2 and SBS13, which are associated with
APOBEC-mediated mutagenesis. Ex vivo drug testing showed broad sensitivity to almost all chemotherapeutic agents
tested. The molecular profile of this patient, characterized by activation of the PI3K pathway and high APOBEC activity,
may contribute to increased susceptibility to cytotoxic agents.
Conclusion: Our results show that the integration of whole-exome sequencing with functional drug profiling could
be a powerful and clinically relevant strategy for personalized diagnostics in NSCLC. This combined approach enables
comprehensive tumor characterization, reveals therapeutic vulnerabilities beyond known driver mutations and
identifies exceptional responders. Such a strategy has the potential to improve patient stratification and make more
effective, individualized treatment decisions in clinical practice.Čavić M, editor. Proceedings book: The 1st Regional SDIR-HDIR-MOKAD Congress; 2025 Oct 8-10; Belgrade, Serbia. Belgrade : Serbian Association for Cancer Research; 2025. p. 66-8. (Oncology Insights; No. 3)