Тесты на лекарственную чувствительность как инструмент коррекции химиотерапии у детей с туберкулезом органов дыхания

Samples from recurrent, treatment-refractory cancers are rarely available, but would be valuable in understanding the molecular drivers of drug resistance. In leukemias, consecutive samples are readily available during treatment. Hence, we explored here the progression of adult acute myeloid leukemias (AML) by serial sampling and by integrating data from multiple platforms. Next-generation exome and RNA sequencing, and phosphoproteomic data were combined with comprehensive 240 cancer drug sensitivity and resistance testing (DSRT) of leukemic blasts ex-vivo before and after clinical relapse. The data were generated in an experimental diagnostic setting, with intent to improve and personalize treatment of patients with recurrent AML. A 54-year old AML-M5 patient with a FLT-3-ITD mutation and a normal karyotype was monitored by serial sampling. The patient was initially refractory to three consecutive high-dose induction treatments and had limited therapy options. AML blasts from the patient were screened with the DSRT platform. Results implied that the blast cells were 710-times more sensitive to temsirolimus and other rapamycin analogs as compared to normal BM cells, and showed a 1100-fold increased sensitivity to dasatinib. Proteomic analysis showed high phosphorylation of several signaling molecules, such as the insulin receptor and mTOR. Sequencing identified WT1 mutations and a NUP98-NSD1 fusion transcript, an infrequent event associated with poor prognosis in AML. Based on the DSRT results, the patient received compassionate off-label treatment with dasatinib, sunitinib and temsirolimus, resulting in a remarkable clinical remission, normalization of blast counts and a rapid recovery of neutrophil counts as a sign of selective elimination of the leukemic cells. The patient relapsed 4 weeks later, and at this point a new DSRT assay was performed, which showed the blast cells to be completely resistant to temsirolimus and less sensitive to dasatinib ex vivo. Consistent with this drug sensitivity profile was a genomic evolution of a distinct AML subclone with new changes, such as NF1 mutation and a microdeletion of the LEF1 gene, which were not observed in the pre-treatment sample. Taken together, we have demonstrated, how molecular profiling and functional ex vivo drug sensitivity and resistance data can be used to individually optimize patient treatment. Remission was achieved in a patient with advanced, treatment-refractory AML. Serial sampling from human AML patients coupled with molecular profiling and drug sensitivity testing may shed light on clonal progression of disease, and the molecular events underlying drug response. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4580. doi:1538-7445.AM2012-4580

Objective To investigate the adhesion levels in uropathogenic Escherichia coli with various degree of drug resistance. Methods One hundred strains of Escherichia coli isolated from urine specimen were collected from patients admitted to 4 Grade A tertiary hospitals in Tianjin during March 2012 to October 2015. Escherichia coli were divided into drug sensitive group and drug resistant group by drug sensitivity tests with 50 strains in each group. The expressions of fimH, fimA, fimB genes of type I fimbriae and papA, papB, papC, papGII genes of P fimbriae were detected by polymerase chain reaction(PCR) and real-time fluorescence quantitative RCR (RT-PCR), respectively. Adhesion ability of type I fimbriae and P fimbriae were tested by yeast cell adhesion test and erythrocyte agglutination test. Chi square test and t(Z) test were used to analyze the data. Results The positive rate of papGII in drug resistant group (42.0%) was significantly higher than that in the drug sensitive group (16.0%) (χ2=8.208, P 0.05). The expression levels of fimH, fimB and papC genes in the sensitive group were higher than those in the resistant group(Z=3.427, t=5.182 and 8.120, all P<0.05). The adhesion ability of strains carrying type I fimbriae in sensitive group was stronger than that of resistant group(χ2=5.769, P<0.05). Conclusions The decline in adhesion ability of type I fimbriae in drug resistant E. coli strains is possibly associated with the adaptive cost of bacteria, the transcription and deficiency of other genes encoded by fim and pap gene cluster will also affect the adhesion function of type I pili and type P pili. Key words: Uropathogenic Escherichia coli; Drug resistance; Adhesion; Fimbriae, bacterial

Objective To explore the pathogenic bacteria distribution and clinical characteristics of ventilator associated pneumonia(VAP) in patients with chronic obstructive pulmonary disease(COPD). Methods 54 COPD patients with respiratory failure were selected as the research subjects, they were taken endotracheal intubation or cut operation mechanical ventilation for treatment, for the specimens collected in the lower respiratory tract secretions or bronchoalveolar lavage fluid, bacteria isolation, culture and drug sensitivity test, and the pathogen distribution, drug resistance and clinical characteristics were analyzed. Results 82 strains of pathogens were selected in 54 specimens, including gram negative bacteria 54 strains(65.85%), common with Pseudomonas aeruginosa(24.39%), Acinetobacter baumannii Acinetobacter rare(17.07%); gram positive bacteria 19 strains(23.17%), Staphylococcus aureus(12.20%) was more common; 9 strains of fungi (10.98%), with candida albicans(7.32%). Meropenem, imipenem, amikacin, ciprofloxacin, cefoperazone/sulbactam resistance rate of Pseudomonas aeruginosa was low and Acinetobacter bacteria to meropenem, imipenem, cefoperazone/sulbactam aztreonam, levofloxacin resistance rate was low, pneumonia Cray Borrelia bacteria to meropenem, imipenem and levofloxacin, ciprofloxacin resistance rate was low.The sensitivity of Staphylococcus aureus to vancomycin and linezolid without resistance to teicoplanin, imipenem resistant rate was low.The mortality rates of ventilator less than 4h and more than 4h were 56.25%, 43.75%, the difference was not statistically significant (χ2=2.64, P>0.05), but the mortality rate of de-escalation therapy was 19.35%, which was significantly lower than 43.48% of rose escalation therapy(χ2=8.54, P<0.05). Conclusion Gram negative bacteria infection in COPD patients combined with VAP is the main infection and pathogens to antimicrobial drug resistance higher.Therefore, the control of nosocomial infection should be strengthen, according to the etiology for the rational use of antimicrobial drugs, and clinical de-escalation therapy can reduce the mortality. Key words: Pulmonary disease, chronic obstructive; Pneumonia, ventilator-associated; Pathogen; Bacteria

Main objective is to explore the characteristics, drug resistance, and risk factors of pathogens in urinary tract infection patients complicated with urinary calculi. A total of 417 patients with urinary calculi who were treated in our hospital from January 2017 to December 2019 were enrolled in this study, including 234 patients with urinary tract infection and 183 patients without urinary tract infection. The midstream urine of 234 patients with urinary tract infection were cultured and used in the drug sensitivity test. Univariate and multivariate logistic regression analysis was used to analyze the risk factors of postoperative urinary tract infection in patients with urinary calculi. A total of 624 strains of pathogens were isolated from the urine samples of 234 patients with postoperative urinary tract infection, of which 386 strains were gram-negative bacteria (61.86 %), 169 strains were gram-positive bacteria (27.08 %), and 69 strains were fungus (11.06 %). In the drug resistance test, the resistance rates of Staphylococcus aureus to tetracycline, rifampicin, and levofloxacin, and the resistance rate of Enterococcus faecium to levofloxacin was low. Enterococcus faecalis had the lowest resistance rate to levofloxacin, followed by azithromycin and gentamicin. The resistance rates of Staphylococcus epidermidis to levofloxacin and vancomycin were low and the resistance rate to erythromycin was 100.00 %. The resistance rate of Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa, and Proteus mirabilis to meropenem and amikacin, and the resistance rate of Klebsiella pneumoniae and Proteus mirabilis to etilmicin and ceftriaxone were low. Univariate analysis showed that the postoperative urinary tract infection in patients with urinary calculi was related to the age, operation time, postoperative indwelling catheter time, stone size, and preoperative prophylactic use of antibiotics. Multivariate logistic regression analysis showed that the age (≥65 y old), operation time (≥60 min), postoperative indwelling catheter time (≥7 d), and no preoperative prophylactic use of antibiotics were the risk factors of postoperative urinary tract infection in patients with urinary calculi. Patients with urinary calculi were prone to urinary tract infection after the operation. The main pathogens of infection were gram-negative bacteria and gram-positive bacteria, and they were resistant to some antibiotics. Antibiotics should be treated reasonably according to the risk factors of urinary tract infection and the results of the drug sensitivity tests to avoid further aggravation of bacterial drug resistances.

ObjectiveTo evaluate the clinical value of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) in detecting Nontuberculous mycobacteria (NTM).MethodsThe clinical data of 172 patients with suspected NTM lung disease were collected from our hospital from January 1, 2018, to December 30, 2021. The results were compared with those of BACTEC MGIT 960 in liquid culture and gene chip. This study also utilised MALDI-TOF MS to detect macrolide (MA) and amikacin (Am) mutations.ResultsOne hundred thirty-seven cases of NTM pulmonary disease were confirmed by identifying the NTM gene chip in bronchoalveolar lavage fluid and/or MALDI-TOF MS detection. The positive predictive value and negative predictive value were 100% (131/131) and 85.37% (35/41), respectively, and the consistency of the two methods was high (kappa=0.899). For the drug resistance detection of MAs, the consistency rate between MALDI-TOF MS detection and drug sensitivity detection was 97.71% (128/131), the sensitivity was 81.25% (13/16) and the specificity was 100% (115/115). The positive and negative predictive values were 100% (13/13) and 93.75% (115/118), respectively. There was no coincidental consistency between the two methods, and the consistency was high (P<0.001, kappa=0.884). For the drug resistance test of Am, the consistency rate between the MALDI-TOF MS test and the drug sensitivity test was 93.13% (122/131), the sensitivity was 93.52% (101/108), the specificity was 90.91% (21/23) and the positive predictive value and negative predictive value were 98.06% (101/103) and 75.00% (21/28), respectively. The two methods had high consistency, and the consistency was not coincidental (P<0.001, kappa=0.781).ConclusionUtilising MALDI-TOF MS has a good consistency with the drug resistance gene chip method and can be a rapid and effective method to identify strains and drug resistance of NTM. Therefore, it has certain clinical application value in patients with suspected NTM lung disease.

To study sensitivity of drug resistance indexes and resistance manner in acute myeloid leukemia (AML), MTT drug sensitivity, growth types of CFU-L in vitro, Bcl-2 antigen and Bcl-2/Bax ratio and intracellular fluorescence intensity of daunorubicin (DNR) were determined. In 62 cases of AML, the positive coincidence rate was 73% with MTT test and the negative coincidence rate was 70%. In 3 commonly used drugs, if one drug showed sensitivity, the coincidence remission rate reached 71%. In 51 cases of AML, there were 31 patients in the group of complete remission (CR), in which CFU-L of 29 patients showed independent growth. CFU-L of 2 patients showed no growth. However, there were 20 patients in the group of non-remission (NR), in which CFU-L of 14 patients showed independent growth. CFU-L of 6 patients showed non-growth pattern. Statistical analysis showed significant difference (P < 0.05). In 32 cases of AML, the expression rate of Bcl-2 was 59.55% +/- 19.56% in drug-sensitive group, and one was 77.36% +/- 11.91% in drug-resistant group, respectively (P < 0.05). At the same time, the ratio of Bcl-2/Bax was 7.50 +/- 5.04 in drug-sensitive group and one was 14.32 +/- 8.99 in drug-resistant group, respectively (P < 0.05). In 15 case of clinically drug-resistant AML, the fluorescence histogram of DNR showed left-shift of main peak (LSMP) in 12 patients. They were diagnosed as classical drug resistance. Meanwhile, 1 patient showed right-shift of main peak (RSMP) in 3 patients. They were diagnosed as re-growth drug resistance. It is concluded that MTT and CFU-L might be used for prediction of drug sensitivity or resistance when patients were on treatment. Bcl-2 and ratio of Bcl-2/Bax might be associated with the prognosis. DNR histogram could be employed for identify the pattern drug resistance. The strength and weakness of these techniques were discussed.

Objective: The gold standard for pulmonary tuberculosis diagnosis is the demonstration of Mycobacterium tuberculosis bacilli. Drug susceptibility test results of the bacilli obtained are crucial in tuberculosis treatment management. The various tests used to determine drug susceptibility must yield the same result. In the present study, we aimed to evaluate the compatibility of drug susceptibility test results obtained by Löwenstein-Jensen (L-J) and BACTEC 460TB methods from the past to the present. Material and Methods: Sputum results from 79 patients suspected of multidrug-resistant pulmonary tuberculosis (MDR-TB) clinically, radiologically, and bacteriologically were evaluated for isoniazid, rifampicin, ethambutol, and streptomycin between June 1997 and June 1998. Culture and drug sensitivity tests on the L-J medium were conducted at the Heybeliada Chest Diseases and Thoracic Surgery Center bacteriology laboratory, while culture and drug sensitivity tests with the BACTEC 460 TB system were performed at another center. The results were assessed for compatibility using the Kappa (κ) test, a tool for comparing two independent parameters. Results: All drug sensitivity tests for isoniazid, rifampicin, ethambutol, and streptomycin were collectively evaluated. It was determined that 263 (83.3%) of 316 drug sensitivity tests yielded concordant results, while 53 (16.7%) produced discordant results. The drug sensitivity tests using L-J and BACTEC 460TB methods indicated compatibility only for streptomycin (κ = 0.715). In contrast, they yielded different results for isoniazid (κ = 0.585), ethambutol (κ = 0.552), and rifampicin (κ = 0.507). Streptomycin exhibited compatibility, while isoniazid, ethambutol, and rifampicin showed incompatibility between the L-J and BACTEC 460TB methods. Conclusion: Drug sensitivity tests are pivotal in tuberculosis treatment management. While literature suggests compatibility between L-J and BACTEC 460TB methods, our study revealed incompatibility. Evaluation of drug sensitivities may lead to confusing results. Current practices involving studies in the same laboratory and genetic testing contribute to faster and more accurate outcomes in managing drug-resistant tuberculosis. Genetic tests and reference laboratories remain crucial for antituberculosis drug sensitivity, emphasizing their continued importance.

Due to the abuse of antibiotics in clinical, animal husbandry, and aquaculture, drug-resistant pathogens are produced, which poses a great threat to human and the public health. At present, a rapid and effective drug sensitivity test method is urgently needed to effectively control the spread of drug-resistant bacteria. Using methylene blue as a redox probe, the electrochemical signals of methylene blue in drug-resistant Escherichia coli strains were analyzed by a CV method. Graphene ink has been used for enhancing the electrochemical signal. Compared with the results of the traditional drug sensitivity test, we proposed a rapid electrochemical drug sensitivity test method which can effectively identify the drug sensitivity of Escherichia coli. The sensitivity of four E. coli isolates to ciprofloxacin, gentamicin, and ampicillin was tested by an electrochemical drug sensitivity test. The respiratory activity value %RA was used as an indicator of bacterial resistance by electrochemical method.

Acute myeloid leukemia (AML) is an aggressive, heterogeneous disease with poor survival after disease recurrence. Although new targeted therapies have recently been approved for specific AML subtypes, the majority is treated with conventional cytotoxic therapy with variable outcome. To identify novel therapies, we performed comprehensive ex vivo drug sensitivity testing with 515 drugs and RNA sequencing on 127 AML patient samples, allowing us to identify associations between transcriptomic profiles and drug responses. Bone marrow or peripheral blood mononuclear cells (MNCs) were collected from diagnostic (n=66), relapsed (n=39) and refractory (n=22) AML patients. RNA was prepared, sequenced and analyzed as described previously (PMID:28818039). Drug sensitivity and resistance testing was performed on the MNCs with 515 approved and investigational oncology chemical compounds (PMID:24056683). To identify expression profiles associated with drug response, generalized linear regression and elastic net regression models were applied. In the analysis, confounding factors including the patient's gender, RNA extraction and library preparation methods were taken into account. Elastic net regression analysis resulted in significant (FDR&amp;lt;0.1) positive or negative associations between 1110 genes and 105 drugs. Clustering of the genes depicted 4 major hubs where drugs with the same mode of action grouped together, e.g. chemotherapeutics, BCL-2, and FLT3 inhibitors. Functional enrichment analysis of each hub using DAVID tool revealed genes involved in regulation of cell proliferation (43 genes, FDR=9.19E-18), which can be explained by the cytotoxic chemotherapy drugs. Genes involved in cell death (68 genes, FDR=1.67E-5) were associated with BCL-2 inhibitor response, while genes involved in cell surface receptor linked signal transduction (51 genes, FDR=2.33E-11) were associated with FLT3 inhibitors. The fourth hub was enriched with cell adhesion (45 genes, FDR=1.11E-21) and focal adhesion (50 genes, FDR=5.11E-28), specifically integrin family (29 genes, FDR=3.17E-18) genes, that play an important role in drug resistance and were negatively correlated with cytotoxic drugs. Finally, the linear regression analysis revealed significant positive correlation between tyrosine kinase inhibitors (sorafeninb, sunitinib, tivozanib) and FLT3LG and KITLG and negative correlation with BEX2 and BEX5 genes. Regression analysis for MEK inhibitors resulted in expected positive correlation with RRAS and JAK2 gene expression. In conclusion, identifying associations between transcriptomic profiles and drug responses may reveal clinically actionable drugs for AML patients characterized by specific molecular features. Our results indicate potential gene expression biomarkers for key targeted drugs, which can be used to identify AML patients likely to benefit from these therapies. Citation Format: Ashwini Kumar, Disha Malani, Bhagwan Yadav, Mika Kontro, Matti Kankainen, Swapnil Potdar, Simon Anders, Kimmo Porkka, Olli Kallioniemi Kallioniemi, Caroline Heckman. Transcriptomic features predicting drug sensitivity and resistance in acute myeloid leukemia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 286.

Objective To investigate the results of etiology and condition of susceptibility test of bacterial pneumonia caused by measles in order to provide basis for clinical diagnosis and treatment. Methods From April 2007 to December 2008, qualified sputum specimens of lower respiratory tract from 105 children diagnosed as bacterial pneumonia caused by measles were carried on a germ development analyzed by bacterial culture. According to the clinical medication, curative effects and results of sputum culture, 105 specimens were divided into drug sensitivity group (n=80) and drug resistance group (n=25). Informed consent was obtained from all participates. Results ① Infection types: The infection of Gram-negative bacteria was 92 cases(87.60%), among them, the first three are escherichia coli, Klebsiella pneumonia, and haemophilus parainfluenzae. The infection of Gram-positive bacteria was 8 cases(7.62%), including 6 cases of streptococcus pneumoniae, 1 case of staphylococcus aureus, and 1 case of hemolytic streptococcus. And the fungous infection was 5 cases(4.76%). ②Results of susceptibility test: The drug sensitive rates of Gram-negative bacteria and imipenem were 100.00%, respectively. Aminoglycosides, quinolones and sulbactam also had high drug sensitive rates. The drug sensitive rates of rifampicin (RFP) to Gram-positive bacteria was 100.00%, to vancomycin and ofloxacin were 87.50%, respectively. As for fungi, the drug sensitive rates of fluconazol and amphotericin were 100.00%. ③ Curative effects and prognosis: The ineffective treatment of drug sensitivity group was 19 cases, but after adjusting the therapeutic regimen according to the results of drug sensitive test, 17 cases (89.40%) cured and 2 had no more improvement. The ineffective treatment of drug resistance group was 25 cases, among them, 21 cases (84.00%) recovered after adjusting the therapeutic regimen according to the results of drug sensitive test for 3 days, 2 died of pneumothorax, and 2 discharged without recovery. Conclusion The main pathogenic bacteria of bacterial pneumonia caused by measles is conditional pathogenic bacteria, and hospital onset of infection is another reason, so early detection, diagnosis, and treatment could decrease the bacterial resistance. Key words: measles; bacterial pneumonia; pathogenic bacteria; drug sensitivity

PurposeTo investigate the CT features of drug-resistant pulmonary tuberculosis (DR-PTB) and the diagnostic value of CT in DR-PTB diagnosis to provide imaging evidence for the timely detection of drug-resistant Mycobacterium tuberculosis.Materials and MethodsA total of 1546 cases of pulmonary tuberculosis (PTB) with complete clinical data, chest CT images and defined drug sensitivity testing results were consecutively enrolled; 516 cases of DR-PTB were included in the drug-resistant group, and 1030 cases of drug-sensitive pulmonary tuberculosis (DS-PTB) were included in the drug-sensitivity group. Comparative analyses of clinical symptoms and imaging findings were conducted. Univariate and logistic regression analyses were performed, a regression equation model was developed, and the receiver operating characteristic (ROC) curve was constructed.ResultsIn the univariate analysis, some features, including whole-lung involvement, multiple cavities, thick-walled cavities, collapsed lung, disseminated lesions along the bronchi, bronchiectasis, emphysema, atelectasis, calcification, proliferative lesions, encapsulated effusion, etc., were observed more frequently in the DR-PTB group than in the DS-PTB group, and the differences were statistically significant (p<0.05). Exudative lesions and pneumoconiosis were observed more frequently in the drug-sensitivity group than in the drug-resistant group (p<0.05). Logistic regression analysis indicated that whole-lung involvement, multiple cavities, thick-walled cavities, disseminated lesions along the bronchi, bronchiectasis, and emphysema were independent risk factors for DR-PTB, and exudative diseases were protective factors. The total prediction accuracy of the regression model was 80.6%, and the area under the ROC curve (AUC) was 82.6%.ConclusionChest CT manifestations of DR-PTB had certain characteristics that significantly indicated the possibility of drug resistance in tuberculosis patients, specifically when multifarious imaging findings, including multiple cavities, thick-walled cavities, disseminated lesions along the bronchi, whole-lung involvement, etc., coexisted simultaneously. These results may provide imaging evidence for timely drug resistance detection in suspected drug-resistant cases and contribute to the early diagnosis of DR-PTB.

This study aims to evaluate the diagnostic value of Xpert MTB/ RIF assay in bronchoalveolar lavage fluid (BALF) in subjects with smear-negative pulmonary tuberculosis. From January 2019 to December 2019, 197 patients with suspected pulmonary tuberculosis were recruited, and bronchoalveolar lavage fluid was collected for acid-fast staining smear, liquid culture of Mycobacterium combined drug sensitivity and Xpert MTB/RIF detection. The sensitivity, specificity, positive predictive value and negative predictive value of Xpert MTB/RIF in bronchoalveolar lavage fluid (BALF) were calculated with smear-negative pulmonary tuberculosis as the reference standard. The consistency of xpert MTB/RIF in the diagnosis of rifampicin resistance was evaluated, with the results of Mycobacterium liquid culture drug sensitivity test and drug sensitivity test as the gold standards. The results showed that among 197 suspected tuberculosis patients, 55 patients were not diagnosed with tuberculosis and 142 patients were diagnosed with smear-negative pulmonary tuberculosis. One hundred and twenty three cases (86.62%) were positive for Xpert MTB/ RIF in bronchoalveolar lavage fluid, 15 cases (10.56%) were positive by acid-fast staining smear method, and 88 cases (61.97%) were positive by the liquid culture method. The positive rate of Xpert MTB / RIF was 93.18% (82 / 88), which was higher than that of 75.93% (41 / 54) of the negative BALF mycobacterium culture (χ 2 = 8.598, P&lt;0.01). The sensitivity and specificity of Xpert MTB/RIF for rifampicin resistance were 100.00% and 97.30%, respectively. Therefore, the diagnostic value of Xpert MTB/RIF in bronchoalveolar lavage fluid for bacterialnegative pulmonary tuberculosis is superior to the acid-fast staining smear of lavage fluid and the mycobacterium culture method.

Introduction/Purpose: Cancer therapy is increasingly moving towards individualized care and therapy, but there are still gaps between what is known and described on the molecular level about cancers and what is applied in the clinic. In an attempt to bridge the knowledge gap, we at the Institute for Molecular Medicine Finland (FIMM) have set up an Individualized Systems Medicine program that integrates clinical information, molecular profiling and functional information about individual patients’ cancers (Pemovska et al, Cancer Discov, 2013). Central to this program is the Drug Sensitivity and Resistance Testing (DSRT) where we functionally profile the responses of primary cancer cells to a comprehensive clinical oncology and signal transduction inhibitor drug collection of 528 compounds. Methods: Acoustic dispensing platforms are integral to the success of this profiling activity. We have to date produced approximately 3000 drug sets as dose response assay ready plates. The acoustic dispensing allows for making pre-drugged single drug plate sets and/or drug combination plates within hours after sampling of the cells. The plates are also readily sent to researchers anywhere in the world for running comparable assays at other sites. The drugging reproducibility is excellent generating results with correlations of 0.98 or higher in replicate assays. We have developed in-house software solutions to aid these processes: a script for quick creation of transfer list for combination plates and automated analysis pipelines with web-based software interfaces to enable the screening biologists to analyze the screening results effectively. Results: The results of these assays are used to explore and understand cancer biology in terms of druggability, functional heterogeneity and mechanism of drug response and resistance. The profiling data can be used to stratify and position the relevance of specific drugs in different diseases and has been used to identify novel clinically relevant activities of existing and investigational drugs (see e.g. Pemovska et al, Nature, 2015). This information is further utilized to establish hypotheses on drug combinations selectively targeting individual cancers and their predictive biomarkers, which can be explored in the clinic by our clinical collaborators to guide the treatment of the individual patient. Conclusions: In summary, we describe our platform for a functional drug sensitivity testing within our individualized cancer systems medicine program, which generates consistent biological and clinically relevant data. Citation Format: Sergey G. Kuznetsov, Alexander Ianevski, Evgeny Kulessky, Karoliina Laamanen, Elina Lehtinen, Maria Nurmi, Swapnil Potdar, Jani Saarela, Katja Suomi, Laura Turunen, Krister Wennerberg, Päivi Tammela. Ex vivo drug sensitivity testing of primary cells for precision cancer medicine [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2153.

Introduction/Purpose:Cancer therapy is increasingly moving towards individualized care and therapy, but there are still gaps between what is known and described on the molecular level about cancers and what is applied in the clinic. In an attempt to bridge the knowledge gap, we at the Institute for Molecular Medicine Finland (FIMM) have set up an Individualized Systems Medicine program that integrates clinical information, molecular profiling and functional information about individual patients' cancers (Pemovska et al, Cancer Discov, 2013). Central to this program is the Drug Sensitivity and Resistance Testing (DSRT) where we functionally profile the responses of primary cancer cells to a comprehensive clinical oncology and signal transduction inhibitor drug collection of 528 compounds.Methods:Acoustic dispensing platforms are integral to the success of this profiling activity. We have to date produced approximately 3000 drug sets as dose response assay ready plates. The acoustic dispensing allows for making pre-drugged single drug plate sets and/or drug combination plates within hours after sampling of the cells. The plates are also readily sent to researchers anywhere in the world for running comparable assays at other sites. The drugging reproducibility is excellent generating results with correlations of 0.98 or higher in replicate assays. We have developed in-house software solutions to aid these processes: a script for quick creation of transfer list for combination plates and automated analysis pipelines with web-based software interfaces to enable the screening biologists to analyze the screening results effectively.Results:The results of these assays are used to explore and understand cancer biology in terms of druggability, functional heterogeneity and mechanism of drug response and resistance. The profiling data can be used to stratify and position the relevance of specific drugs in different diseases and has been used to identify novel clinically relevant activities of existing and investigational drugs (see e.g. Pemovska et al, Nature, 2015). This information is further utilized to establish hypotheses on drug combinations selectively targeting individual cancers and their predictive biomarkers, which can be explored in the clinic by our clinical collaborators to guide the treatment of the individual patient.Conclusions:In summary, we describe our platform for a functional drug sensitivity testing within our individualized cancer systems medicine program, which generates consistent biological and clinically relevant data.Citation Format: Sergey G. Kuznetsov, Alexander Ianevski, Evgeny Kulessky, Karoliina Laamanen, Elina Lehtinen, Maria Nurmi, Swapnil Potdar, Jani Saarela, Katja Suomi, Laura Turunen, Krister Wennerberg, Päivi Tammela. Ex vivo drug sensitivity testing of primary cells for precision cancer medicine [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2153.

The heterogeneity of chemoresponses in high-grade serous ovarian cancer (HGS-OvCa) presents a major clinical challenge. The inherited or acquired nonresponsiveness to current therapies is likely to be one reason for relapse and treatment failure. Better models for predicting effective treatment options and drug combinations are needed. Although most tumors initially respond to standard platinum-taxane combination therapy, treatment resistance eventually evolves in 80-90% of the patients. Drug sensitivity and resistance testing (DSRT) was performed by using high-throughput screening (HTS) of 306 small-drug molecules on six primary cell lines from patients with disseminated HGS-OvCa. Commonly available ovarian cancer cell line OVCAR8 was used as a control. The drug effect was investigated with help of a quantitative scoring approach, drug sensitivity score (DSS). Sixteen drugs of clinical interest were investigated in more detail. Drug sensitivity varied greatly within primary cell lines. Out of 306 compounds, 29 were effective on the most resistant cell line, whereas 102 compounds showed effect on the most sensitive line. We found that 9 out of the 16 clinically important compounds gave no response on the tested cell lines. Seven compounds provided response on the screened cell lines: four HSP90 inhibitors, an HSP70 inhibitor, a Wee1 inhibitor, and a proteasome inhibitor. Of these seven compounds, the most effective drug compounds (the Wee1 inhibitor and two HSP90 inhibitors) were chosen for validation on primary HGS-OvCa cell lines to verify the HTS result. The effectiveness of currently used clinical treatment-line drugs, cisplatin and paclitaxel, was compared to the putative novel drug compounds on the primary cell lines and the control cell line. One primary cell line was completely resistant to the validated drug compounds as well as to cisplatin and paclitaxel combination treatment. Five cell lines were sensitive to cisplatin-paclitaxel treatment, but the sensitivity to the three most effective compounds varied greatly. The commonly available cell line OVCAR8 was the most sensitive of all tested cell lines. In conclusion, we found three potential drug compounds of clinical interest effective for disseminated HGS-OvCa. The analyses demonstrate that patient-derived primary HGS-OvCa cell lines have unique heterogeneous characteristics that are likely to have an effect on the choice of best drug candidates for each individual cell line, and furthermore for each patient. Consequently, more combinatorial drug treatment strategies should be validated to find new specific personalized drug treatment strategies. Citation Format: Pia Roering, Piia Mikkonen, Swapnil Potdar, Krister Wennerberg, Johanna Hynninen, Seija Grénman, Annika Auranen, Olli Carpén, Katja Kaipio. Drug sensitivity and resistance testing (DSRT) of clinically important compounds on primary ovarian cancer cell lines. [abstract]. In: Proceedings of the AACR Conference: Addressing Critical Questions in Ovarian Cancer Research and Treatment; Oct 1-4, 2017; Pittsburgh, PA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(15_Suppl):Abstract nr A57.