Drug Sensitivity Testing Research Articles

Bioinformatics analysis of an immunotherapy responsiveness-related gene signature in predicting lung adenocarcinoma prognosis.

Immune therapy has become first-line treatment option for patients with lung cancer, but some patients respond poorly to immune therapy, especially among patients with lung adenocarcinoma (LUAD). Novel tools are needed to screen potential responders to immune therapy in LUAD patients, to better predict the prognosis and guide clinical decision-making. Although many efforts have been made to predict the responsiveness of LUAD patients, the results were limited. During the era of immunotherapy, this study attempts to construct a novel prognostic model for LUAD by utilizing differentially expressed genes (DEGs) among patients with differential immune therapy responses. Transcriptome data of 598 patients with LUAD were downloaded from The Cancer Genome Atlas (TCGA) database, which included 539 tumor samples and 59 normal control samples, with a mean follow-up time of 29.69 months (63.1% of patients remained alive by the end of follow-up). Other data sources including three datasets from the Gene Expression Omnibus (GEO) database were analyzed, and the DEGs between immunotherapy responders and nonresponders were identified and screened. Univariate Cox regression analysis was applied with the TCGA cohort as the training set and GSE72094 cohort as the validation set, and least absolute shrinkage and selection operator (LASSO) Cox regression were applied in the prognostic-related genes which fulfilled the filter criteria to establish a prognostic formula, which was then tested with time-dependent receiver operating characteristic (ROC) analysis. Enriched pathways of the prognostic-related genes were analyzed with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses, and tumor immune microenvironment (TIME), tumor mutational burden, and drug sensitivity tests were completed with appropriate packages in R (The R Foundation of Statistical Computing). Finally, a nomogram incorporating the prognostic formula was established. A total of 1,636 DEGs were identified, 1,163 prognostic-related DEGs were extracted, and 34 DEGs were selected and incorporated into the immunotherapy responsiveness-related risk score (IRRS) formula. The IRRS formula had good performance in predicting the overall prognoses in patients with LUAD and had excellent performance in prognosis prediction in all LUAD subgroups. Moreover, the IRRS formula could predict anticancer drug sensitivity and immunotherapy responsiveness in patients with LUAD. Mechanistically, immune microenvironments varied profoundly between the two IRRS groups; the most significantly varied pathway between the high-IRRS and low-IRRS groups was ribonucleoprotein complex biogenesis, which correlated closely with the TP53 and TTN mutation burdens. In addition, we established a nomogram incorporating the IRRS, age, sex, clinical stage, T-stage, N-stage, and M-stage as predictors that could predict the prognoses of 1-year, 3-year, and 5-year survival in patients with LUAD, with an area under curve (AUC) of 0.718, 0.702, and 0.68, respectively. The model we established in the present study could predict the prognosis of LUAD patients, help to identify patients with good responses to anticancer drugs and immunotherapy, and serve as a valuable tool to guide clinical decision-making.

Analysis of drug resistance in pulmonary tuberculosis patients with positive sputum tuberculosis culture in Northeast China.

Objective: The objective of this study is to determine the drug resistance status of pulmonary tuberculosis patients in Jilin Province. Methods: A retrospective survey was conducted on 395 sputum culture TB-positive patients admitted to the tuberculosis hospital in Jilin Province in 2019. Sputum samples were cultured in acidic Roche medium. Drug sensitivity testing was conducted using the proportional method. Sensitivity was reported if the percentage of drug resistance was less than 1%, and resistance was reported if the percentage was ≥1%. Statistical analysis was performed using SPSS 22.0. Results: 395 tuberculosis patients with positive sputum tuberculosis culture were included in the study, with 102 being initially treated and 293 being retreated. The study population consisted of 283 males and 112 females. Sex, age, nationality, occupation, marital status, diabetes comorbidity, initial treatment, normal health status, BCG vaccine vaccination, smoking, and alcohol consumption were considered as factors that may affect the rate of multidrug resistance. And only the history of treatment (initial treatment) was associated with multidrug resistance (p = 0.032). This indicates that retreatment is the most significant risk factor for the occurrence of multidrug resistance in tuberculosis. The multidrug resistance rate in retreated patients is 3.764 times higher than that in initially treated patients. Conclusion: The prevalence of multidrug-resistant is higher in retreated patients compared to initially treated patients in the study population. Multidrug resistance is only associated with the treatment history (initial retreatment) and not with other factors.

Study on the Effect of Shuanghuanglian Powder Needle Combined with Cefoperazone and Sulbactam Sodium on <i>Pseudomonas aeruginosa in Vitro</i>

Objective: To explore the antibacterial activity of combined use of Shuanghuanglian and cefoperazone sulbactam sodium on resistant strains of Pseudomonas aeruginosa. Methods: The Pseudomonas aeruginosa strains which were sensitive and resistant to cefoperazone sulbactam sodium were selected to prepare different test bacterial solutions respectively; The experimental liquid of Shuanghuanglian and Cefoperazone Sulbactam Sodium were prepared separately and set as different test groups and control groups; The Drug Sensitivity Tests of Shuanghuanglian and cefoperazone sulbactam sodium at different concentration gradients which were used alone or used in combination were carried out for different strains with sensitivity and resistance, And use standard entry as a reference control. Result: The results of drug sensitivity test of Shuanghuanglian combined with Cefoperazone-Sulbactam sodium against the resistant strains of Pseudomonas aeruginosa were compared with the results of drug sensitivity test of the two separately used, and the difference was statistically significant (P Pseudomonas aeruginosa resistant strains were statistically significant compared with the drug sensitivity test results of Shuanghuanglian and Cefoperazone Sulbactam Sodium used separately (P Conclusion: The combination of Shuanghuanglian and cefoperazone sulbactam sodium has synergistic antibacterial or bactericidal effect on Pseudomonas aeruginosa resistant strains.

Analysis of foodborne salmonellosis serotypes and drug resistance in children in Chenzhou City of China from 2017 through 2022.

Salmonella is a significant pathogens of foodborne illness. The widespread use of antibiotics in clinical practice and animal husbandry has resulted in increasing drug resistance of Salmonella. In this study, we examined the serotype distribution and drug resistance of Salmonella in pediatric patients with diarrhea in Chenzhou City to provide a basis for the scientific control and rational use of antibiotics in clinical practice in relation to Salmonellosis. Stool Salmonella spp. were collected from patients younger than 18 years of age who met the definition for foodborne illness at two sentinel hospitals from 2017 through 2022 tested Salmonella, and a descriptive analysis of the epidemiologic characteristics. Salmonella strains isolated from the stool underwent serology and drug-sensitivity tests. The following 14 antibiotics were used for the drug-sensitivity tests: ampicillin (AMP), ampicillin/sulbactam (AMS), cefazolin (CFZ), cefoxitin, cefotaxime, ceftazidime, imipenem (IPM), tetracycline (TET), nalidixic acid, ciprofloxacin, chloramphenicol (CHL), gentamicin, trimethoprim/sulfamethoxazole (SXT), and azithromycin. Samples from 1,263 pediatric with diarrhea, and Salmonella was detected in 221 (17.5%) of these patients. Positive test results were principally observed in the second and third quarters of each year, accounting for 21.1% and 19.6% of the cases, respectively. The infection rates of infants aged less than 12 months and toddlers aged 1-3 years with diarrhea were the highest at 21.3% and 17.8%, respectively. The 221 Salmonella strains were divided into 32 serotypes, of which Salmonella Typhimurium (S. Typhimurium) was the dominant strain (79.2%). The resistance rates to TET (86.9%), AMP (75.6%), AMS (58.4%), CFZ (55.7%), CHL (54.3%), and SXT (45.2%) predominated, and the differences in the drug-resistance rates to 1st-, 2nd-, and 3rd-generation cephalosporins were high (2.3-55.7%). Only 0.9% of the strains were resistant to IPM. The multidrug resistance (MDR) rate was 76.5% (169/221), and 48.9% (108/221) of the strains were resistant to five or more classes of antibiotics, of which the most common drug-resistance profile was AMP-AMS-TET-CHL-CFZ-SXT, accounting for 10.9% of Salmonella strains (24/221). Foodborne salmonellosis tended to occur during the summer and autumn in children, and infants and toddlers were more likely to develop salmonellosis than children in the other age groups. The dominant Salmonella serotype was S. Typhimurium. The drug-resistance rate of the tested strains was high, and the MDR problem was severe. We recommend that in the treatment of salmonellosis, antibiotics be selected rationally based on the drug-resistance status of local Salmonella resistance situation to ensure safety and efficacy.

Preclinical investigation of patient-derived cervical cancer organoids for precision medicine.

Advanced cervical cancer is still difficult to treat and in the case of recurrent cancer, it is desirable to utilize personalized treatment rather than uniform treatment because the type of recurrence is different for each individual. Therefore, this study aimed to establish a patient-derived organoid (PDO) platform to determine the effects of chemotherapy, radiation therapy, and targeted therapy in cervical cancer. We established organoids from 4 patients with various types of cervical cancer. The histopathological and gene profiles of these organoid models were compared to determine their characteristics and the maintenance of the patient phenotype. Each type of organoid was also subjected to anticancer drug screening and radiation therapy to evaluate its sensitivity. We established PDOs to recapitulate the main elements of the original patient tumors, including the DNA copy number and mutational profile. We selected 7 drugs that showed growth inhibition in cervical cancer organoids out of 171 using an Food and Drug Administration -approved drug library. Moreover, adenocarcinoma and large-cell neuroendocrine carcinoma showed resistance to radiation therapy. whereas squamous cell carcinoma and villoglandular carcinoma showed a significant response to radiotherapy. Our results showed that patient-derived cervical cancer organoids can be used as a platform for drug and radiation sensitivity testing. These findings suggest that patient-derived cervical cancer organoids could be used as a personalized medicine platform and may provide the best treatment options for patients with various subtypes of cervical cancer.

Ex vivo Drug Sensitivity Imaging-based Platform for Primary Acute Lymphoblastic Leukemia Cells.

Resistance of acute lymphoblastic leukemia (ALL) cells to chemotherapy, whether present at diagnosis or acquired during treatment, is a major cause of treatment failure. Primary ALL cells are accessible for drug sensitivity testing at the time of new diagnosis or at relapse, but there are major limitations with current methods for determining drug sensitivity ex vivo. Here, we describe a functional precision medicine method using a fluorescence imaging platform to test drug sensitivity profiles of primary ALL cells. Leukemia cells are co-cultured with mesenchymal stromal cells and tested with a panel of 40 anti-leukemia drugs to determine individual patterns of drug resistance and sensitivity ("pharmacotype"). This imaging-based pharmacotyping assay addresses the limitations of prior ex vivo drug sensitivity methods by automating data analysis to produce high-throughput data while requiring fewer cells and significantly decreasing the labor-intensive time required to conduct the assay. The integration of drug sensitivity data with genomic profiling provides a basis for rational genomics-guided precision medicine. Key features Analysis of primary acute lymphoblastic leukemia (ALL) blasts obtained at diagnosis from bone marrow aspirate or peripheral blood. Experiments are performed ex vivo with mesenchymal stromal cell co-culture and require four days to complete. This fluorescence imaging-based protocol enhances previous ex vivo drug sensitivity assays and improves efficiency by requiring fewer primary cells while increasing the number of drugs tested to 40. It takes approximately 2-3 h for sample preparation and processing and a 1.5-hour imaging time. Graphical overview.

Does the Microflora of Surgery Site Infection Change After Prophylactic Use of Vancomycin Powder in the Spine Surgery

Retrospective cohort study. This study aimed to investigate the characteristics of microflora in patients with deep spinal surgical site infection (SSI) after prophylactic use of vancomycin powder (VP). A retrospective analysis was performed on patients after spinal surgery. Patients were grouped according to whether VP use and only patients with deep SSI were included in this study. General information of the patients, the dose of vancomycin, bacterial culture results, drug sensitivity test results, and SSI treatment methods were recorded. The differences of microflora between the two groups were analyzed, and the sensitivity of bacteria in the +VP group to antibiotics was analyzed. The infection rate in the +VP group was 4.9% (56/1124) vs 6.3% (93/1476) in the No-VP group (P < 0.05). The proportion of Gram-positive bacteria (GPB) in the +VP SSIs was 55.4% vs.74.1% in the No-VP group (P < 0.05). The percentage of Gram-negative bacteria (GNB) in the +VP SSIs was 46.4% vs.30.1% in the No-VP group (P < 0.05). More dose of VP cannot decrease the SSI, but the proportion of GNB in VP >1g SSIs was higher (59.0% vs 32.4%, P < 0.05). In the +VP SSIs, all of the GNB cultured were sensitive to meropenem, and linezolid covered most of the GPB cultured. Local use of vancomycin powder can reduce the incidence of SSI, but this may lead to changes in the bacterial flora. Once the SSI occurs, the case of GNB infection may be increased. The more dose of VP cannot decrease SSI but may increase the rate of GNB in the +VP SSIs. Once infections still occur after VP use, antibiotics covering GNB may be added. These findings may help guide choice of empiric antibiotics while awaiting culture data.

Retracted: Correlation between Sputum Bacterial Culture Positive Rate and Drug Sensitivity Test Results and Disease Severity in Inpatients and Its Clinical Significance: A Systematic Review and Meta-Analysis.

[This retracts the article DOI: 10.1155/2022/5102100.].

Screening and Drug Resistance Analysis of Non-Tuberculous Mycobacteria in Patients with Suspected Pulmonary Tuberculosis on the Hainan Island, China

China has seen a drastic increase in the incidence of non-tuberculous mycobacteria (NTM) infection, which is a notable public health issue. Due to a lack of reliable epidemiological surveillance information, there is a need to gather accurate epidemiological and surveillance data, which can help clinicians effectively treat NTM patients. Moreover, drug susceptibility testing for NTM is not frequently performed in China. This retrospective study, therefore, determined the prevalence and resistance characteristics of NTM to provide a reference to control the NTM epidemic. Sputum, alveolar lavage fluid, and other respiratory specimens were collected from 3025 patients with suspected pulmonary tuberculosis attending The Second Affiliated Hospital of Hainan Medical University from January 2014 to December 2021. Strain identification and species distribution of NTM were performed by DNA chip technology and gene sequencing, and the drug resistance of NTM isolates was evaluated by calculating the minimum inhibitory concentration through antimicrobial susceptibility testing for NTM. From 2014 to 2021, 373 strains of NTM were isolated and identified from respiratory specimens of 3025 suspected tuberculosis patients. Except in 2014, NTM-infected patients accounted for more than 10% of suspected tuberculosis patients in other years. The median age of patients with NTM infection was 62.0 years (53.0, 71.0), and the male-to-female ratio among these patients was 0.79:1. Among culture-positive strains, 12.3% (373/3040; 95% CI 11.1-13.4%) were identified as NTM comprising forty species of NTM. The forty species of NTM included 23 slow-growing mycobacteria (SGM) and 17 rapidly-growing mycobacteria (RGM). Among the NTM isolates, 58.7% (219/373; 95% CI 53.7-63.7%) were SGM and 41.3% (154/373; 95% CI 36.3-46.3%) were RGM. M.avium complex(MAC)(41.3%; 95% CI 36.3-46.3%) and M.abscessus complex (MABC)(33.2%; 95% CI 28.4-38.0%) were the most frequently detected species, followed by M.simiae Complex (11.8%; 95% CI 8.5-15.1%), M.fortuitum group (5.1%; 95% CI 2.9-7.3%), and others. Drug sensitivity test results showed that most of the NTM isolates were susceptible to amikacin and clarithromycin with a drug resistance rate of less than 10%. However, clarithromycin could induce drug resistance, followed by linezolid and moxifloxacin, and their drug resistance rate was less than 50%. During 2014-2021, the number of NTM isolates detected in the respiratory specimens of the study patients in The Second Affiliated Hospital of Hainan Medical University increased year by year. M. intracellulare is the most common pathogenic NTM species, and there is a high incidence of NTM infection on Hainan Island. Our findings might be of great importance for diagnosing and treating this patient population in Hainan.

Recent progress of nanostructure-based enrichment of circulating tumor cells and downstream analysis.

The isolation and detection of circulating tumor cells (CTCs) play an important role in early cancer diagnosis and prognosis, providing easy access to identify metastatic cells before clinically detectable metastases. In the past 20 years, according to the heterogeneous expression of CTCs on the surface and their special physical properties (size, morphology, electricity, etc.), a series of in vitro enrichment methods of CTCs have been developed based on microfluidic chip technology, nanomaterials and various nanostructures. In recent years, the in vivo detection of CTCs has attracted considerable attention. Photoacoustic flow cytometry and fluorescence flow cytometry were used to detect CTCs in a noninvasive manner. In addition, flexible magnetic wire and indwelling intravascular non-circulating CTCs isolation system were developed for in vivo CTCs study. In the aspect of downstream analysis, gene analysis and drug sensitivity tests of enriched CTCs were developed based on various existing molecular analysis techniques. All of these studies constitute a complete study of CTCs. Although the existing reviews mainly focus on one aspect of capturing CTCs study, a review that includes the in vivo and in vitro capture and downstream analysis study of CTCs is highly needed. This review focuses on not only the classic work and latest research progress in in vitro capture but also includes the in vivo capture and downstream analysis, discussing the advantages and significance of the different research methods and providing new ideas for solving the heterogeneity and rarity of CTCs.

Screening the clinical isolates of Acinetobacter baumannii for biofilm potential/trait from a tertiary care hospital in India

Background: The multidrug resistance and biofilm potential of Acinetobacter baumannii have made it difficult for the clinicians and healthcare providers to treat and control its spread, leading to mortalities: 23% in hospitalized patients and 43% among patients in intensive care. Biofilm has become an important virulent factor for Acinetobacter baumannii as it not only protects the pathogen against the antibiotics but also enables it to evade the immune system of the host. Resistance gene transfer happens among the species and across other species in the biofilm. Knowing if the clinical isolate of A. baumannii from a patient has biofilm trait will help the clinician to make an informed, better antibiotic strategy. The relationship between biofilm trait and multidrug resistance in the clinical isolates of A. baumannii was investigated in this study. Methods: 72 clinical isolates of A. baumannii were included in this study. Antimicrobial drug sensitivity testing was performed on these isolates against two front line antibiotics: Imipenem (10µg) and Meropenem (10µg) by Kirby-Bauer disk diffusion method. Biofilm formation was monitored and estimated quantitatively in 96-well polystyrene plate. The biofilms were examined under scanning electron microscope (SEM) while some key regulatory genes of the biofilm synthetic pathway were screened by PCR to corroborate the phenotype. Results: The 72 clinical isolates were characterized for carbapenem sensitivity. They were grouped as Carbapenem Resistant (CRAB) (n=58) and Carbapenem Sensitive (CSAB) (n=14) isolates. All isolates formed biofilm in our study and based on the amount of biofilm formed they were grouped as: strong biofilm producers 83% (60/72), moderate biofilm producers 11% (08) and weak biofilm producers 6% (04). Genotyping by PCR showed certain key regulatory genes like ata, bap and csu in almost all the isolates. Morphological studies showed that the biofilms were multi-layered, closely knit and with a thick strong matrix clearly visible under SEM. Conclusions: All the isolates efficiently formed robust biofilms, 83% of which formed strong biofilms (P value < 0.05). Both CRAB and CSAB formed biofilms inferring that biofilms production is inherent trait and phenotype of these clinical isolates of A. baumannii. The genotyping confirmed the presence of certain key genes of biofilm pathway and the strong integrity of the biofilms were revealed under SEM. Our study endorses the view that biofilms constitute an inherent trait which apparently enables A. baumannii to acquire MDR phenotype.

Treatment Strategies for Multiple Myeloma Treatment and the Role of High-Throughput Screening for Precision Cancer Therapy.

In the past few years, development of approved drug candidates has improved the disease management of multiple myeloma (MM). However, due to drug resistance, some of the patients do not respond positively, while some of the patients acquire drug resistance, thereby these patients eventually relapse. Hence, there are no other therapeutic options for multiple myeloma patients. Therefore, this necessitates a precision-based approach to multiple myeloma therapy. The use of patient's samples to test drug sensitivity to increase efficacy and reduce treatment-related toxicities is the goal of functional precision medicine. Platforms such as high-throughput-based drug repurposing technology can be used to select effective single drug and drug combinations based on the efficacy and toxicity studies within a time frame of couple of weeks. In this article, we describe the clinical and cytogenetic features of MM. We highlight the various treatment strategies and elaborate on the role of high-throughput screening platforms in a precision-based approach towards clinical treatment.

Challenges Faced by Health Facilities in Implementing Antimicrobial Stewardship Program: A Narrative Review

Background: Antimicrobial Resistance (AMR) has become a significant world-wide health problem. Antimicrobial use though necessary, overusing/misusing can have detrimental long-term effects. Managing AMR involves a lot of challenges, therefore appropriate strategies must be developed to overcome them. Methods: We conducted a comprehensive search using web-based search engines employing a combination of search terms. The search yielded 404 papers between 2000 and 2021. We took up Forty-four papers fulfilling the inclusion-exclusion criteria for review. Results: The challenges identified from studies in implementing the Antimicrobial Stewardship Program (AMSP) include the administration’s lack of knowledge, a deficit of resources, staff turnover, resistance to change, delay in Culture and Drug Sensitivity Testing (CDST), inadequate training, patient expectations, and insufficient Electronic Medical Records (EMR). Conclusion: Antimicrobial resistance is a growing public health concern that all stakeholders must address. Improvements in antimicrobial consumption reporting and locally tailored guidelines can help to ensure successful implementation.

Rapid anticancer drug screening using patient-derived lung cancer organoids: a case report

A variety of anticancer drugs and targeted agents for lung cancer have been developed, but some patients do not respond to these medications as intended. Therefore, there is an urgent need to develop a tool for predicting the anticancer drug response of each patient. Patient-derived tumor organoids (PDOs) have emerged as reliable in vitro tools for developing precision medicine. Herein, we describe a case of a 50-year-old nonsmoking man who was diagnosed with non-small cell lung cancer. Initially, no clinical symptoms were found in this patient. Postoperative pathology confirmed a stage Ib tumor, and an epidermal growth factor receptor (EGFR) mutation (exon21p.L858R) was detected in the patient’s lung tumor specimen. The patient later showed intracranial relapse 17 months after complete resection. An organoid culture was established from the resected brain metastatic tissue and a drug sensitivity test showed, within 72 hours, that the organoids were resistant to gefitinib and osimertinib. Our results recapitulated the patient’s response to anticancer drugs, demonstrating the potential of PDOs for precision medicine.

Multi-omics analyses of tumor-associated immune-infiltrating cells with the novel immune checkpoint protein tyrosine phosphatase 1B (PTP1B) in extracellular matrix of brain-lower-grade-glioma (LGG) and uveal-melanoma (UVM).

Immune checkpoint inhibitors represented by PD-1 have greatly changed the way cancer is treated. In addition to PD-1, new immune checkpoints are constantly excavated to better treat cancer. Recently, protein tyrosine phosphatase 1B (PTP1B) was identified as a new immune checkpoint and played a critical role in the treatment of tumors by inhibiting the proliferation and cytotoxicity of T cells induced by tumor antigen. To explore the targeting role of PTP1B in precision tumor therapy, we deeply analyzed the expression and prognosis of PTP1B in all tumors. Survival analysis results indicated that PTP1B was highly expressed in most tumor tissues and indicated poor prognosis in acute-myeloid-leukemia (LAML), brain-lower-grade-glioma (LGG), kidney-renal clear-cell-carcinoma (KIRC) and uveal-melanoma (UVM). The methylation status of PTP1B in these four tumors exhibited hypomethylation and mutation landscape showed that PTP1B had its specific characteristics in genomic instability and heterogeneity. The homologous recombination deficiency (HRD) and loss of heterozygosity (LOH) were positive related to PTP1B expression in liver-hepatocellular-carcinoma (LIHC) and kidney-chromophobe (KICH), while the immunescore and immune infiltration displayed a significant positive correlation with PTP1B expression in LGG and UVM. Drug sensitivity tests showed that the PTP1B inhibitor MSI-1436 had a sensitivity effect suppressing tumor cell viability and suggested it enhanced the efficacy of PD-1 inhibitors in cancers.

Patient-Derived Organoids: The Beginning of a New Era in Ovarian Cancer Disease Modeling and Drug Sensitivity Testing.

Ovarian cancer (OC) is the leading cause of death from gynecological malignancies. Despite great advances in treatment strategies, therapeutic resistance and the gap between preclinical data and actual clinical efficacy justify the necessity of developing novel models for investigating OC. Organoids represent revolutionary three-dimensional cell culture models, deriving from stem cells and reflecting the primary tissue's biology and pathology. The aim of the current review is to study the current status of mouse- and patient-derived organoids, as well as their potential to model carcinogenesis and perform drug screenings for OC. Herein, we describe the role of organoids in the assessment of high-grade serous OC (HGSOC) cells-of-origin, illustrate their use as promising preclinical OC models and highlight the advantages of organoid technology in terms of disease modelling and drug sensitivity testing.

Ex Vivo Drug Sensitivity Correlates with Clinical Response and Supports Personalized Therapy in Pediatric AML.

Acute myeloid leukemia (AML) is a heterogeneous disease that accounts for ~20% of all childhood leukemias, and more than 40% of children with AML relapse within three years of diagnosis. Although recent efforts have focused on developing a precise medicine-based approach towards treating AML in adults, there remains a critical gap in therapies designed specifically for children. Here, we present ex vivo drug sensitivity profiles for children with de novo AML using an automated flow cytometry platform. Fresh diagnostic blood or bone marrow aspirate samples were screened for sensitivity in response to 78 dose conditions by measuring the reduction in leukemic blasts relative to the control. In pediatric patients treated with conventional chemotherapy, comprising cytarabine, daunorubicin and etoposide (ADE), ex vivo drug sensitivity results correlated with minimal residual disease (r = 0.63) and one year relapse-free survival (r = 0.70; AUROC = 0.94). In the de novo ADE analysis cohort of 13 patients, AML cells showed greater sensitivity to bortezomib/panobinostat compared with ADE, and comparable sensitivity between venetoclax/azacitidine and ADE ex vivo. Two patients showed a differential response between ADE and bortezomib/panobinostat, thus supporting the incorporation of ex vivo drug sensitivity testing in clinical trials to further evaluate the predictive utility of this platform in children with AML.

Erythroid/megakaryocytic differentiation confers BCL-XL dependency and venetoclax resistance in acute myeloid leukemia

AbstractMyeloid neoplasms with erythroid or megakaryocytic differentiation include pure erythroid leukemia, myelodysplastic syndrome with erythroid features, and acute megakaryoblastic leukemia (FAB M7) and are characterized by poor prognosis and limited treatment options. Here, we investigate the drug sensitivity landscape of these rare malignancies. We show that acute myeloid leukemia (AML) cells with erythroid or megakaryocytic differentiation depend on the antiapoptotic protein B-cell lymphoma (BCL)-XL, rather than BCL-2, using combined ex vivo drug sensitivity testing, genetic perturbation, and transcriptomic profiling. High-throughput screening of >500 compounds identified the BCL-XL–selective inhibitor A-1331852 and navitoclax as highly effective against erythroid/megakaryoblastic leukemia cell lines. In contrast, these AML subtypes were resistant to the BCL-2 inhibitor venetoclax, which is used clinically in the treatment of AML. Consistently, genome-scale CRISPR-Cas9 and RNAi screening data demonstrated the striking essentiality of BCL-XL-encoding BCL2L1 but not BCL2 or MCL1, for the survival of erythroid/megakaryoblastic leukemia cell lines. Single-cell and bulk transcriptomics of patient samples with erythroid and megakaryoblastic leukemias identified high BCL2L1 expression compared with other subtypes of AML and other hematological malignancies, where BCL2 and MCL1 were more prominent. BCL-XL inhibition effectively killed blasts in samples from patients with AML with erythroid or megakaryocytic differentiation ex vivo and reduced tumor burden in a mouse erythroleukemia xenograft model. Combining the BCL-XL inhibitor with the JAK inhibitor ruxolitinib showed synergistic and durable responses in cell lines. Our results suggest targeting BCL-XL as a potential therapy option in erythroid/megakaryoblastic leukemias and highlight an AML subgroup with potentially reduced sensitivity to venetoclax-based treatments.

Ex vivo venetoclax sensitivity testing predicts treatment response in acute myeloid leukemia.

The BCL-2 inhibitor venetoclax has revolutionized the treatment of acute myeloid leukemia (AML) in patients not benefiting from intensive chemotherapy. Nevertheless, treatment failure remains a challenge, and predictive markers are needed, particularly for relapsed or refractory AML. Ex vivo drug sensitivity testing may correlate with outcomes, but its prospective predictive value remains unexplored. Here we report the results of the first stage of the prospective phase II VenEx trial evaluating the utility and predictiveness of venetoclax sensitivity testing using different cell culture conditions and cell viability assays in patients receiving venetoclax-azacitidine. Participants with de novo AML ineligible for intensive chemotherapy, relapsed or refractory AML, or secondary AML were included. The primary endpoint was the treatment response in participants showing ex vivo sensitivity and the key secondary endpoints were the correlation of sensitivity with responses and survival. Venetoclax sensitivity testing was successful in 38/39 participants. Experimental conditions significantly influenced the predictive accuracy. Blast-specific venetoclax sensitivity measured in conditioned medium most accurately correlated with treatment outcomes; 88% of sensitive participants achieved a treatment response. The median survival was significantly longer for participants who were ex vivo-sensitive to venetoclax (14.6 months for venetoclax-sensitive patients vs. 3.5 for venetoclax-insensitive patients, P<0.001). This analysis illustrates the feasibility of integrating drug-response profiling into clinical practice and demonstrates excellent predictivity. This trial is registered with ClinicalTrials.gov identifier: NCT04267081.

Impact of molecular diagnostic tests on diagnostic and treatment delays in tuberculosis: a systematic review and meta-analysis

BackgroundCountries with high TB burden have expanded access to molecular diagnostic tests. However, their impact on reducing delays in TB diagnosis and treatment has not been assessed. Our primary aim was to summarize the quantitative evidence on the impact of nucleic acid amplification tests (NAAT) on diagnostic and treatment delays compared to that of the standard of care for drug-sensitive and drug-resistant tuberculosis (DS-TB and DR-TB).MethodsWe searched MEDLINE, EMBASE, Web of Science, and the Global Health databases (from their inception to October 12, 2020) and extracted time delay data for each test. We then analysed the diagnostic and treatment initiation delay separately for DS-TB and DR-TB by comparing smear vs Xpert for DS-TB and culture drug sensitivity testing (DST) vs line probe assay (LPA) for DR-TB. We conducted random effects meta-analyses of differences of the medians to quantify the difference in diagnostic and treatment initiation delay, and we investigated heterogeneity in effect estimates based on the period the test was used in, empiric treatment rate, HIV prevalence, healthcare level, and study design. We also evaluated methodological differences in assessing time delays.ResultsA total of 45 studies were included in this review (DS = 26; DR = 20). We found considerable heterogeneity in the definition and reporting of time delays across the studies. For DS-TB, the use of Xpert reduced diagnostic delay by 1.79 days (95% CI − 0.27 to 3.85) and treatment initiation delay by 2.55 days (95% CI 0.54–4.56) in comparison to sputum microscopy. For DR-TB, use of LPAs reduced diagnostic delay by 40.09 days (95% CI 26.82–53.37) and treatment initiation delay by 45.32 days (95% CI 30.27–60.37) in comparison to any culture DST methods.ConclusionsOur findings indicate that the use of World Health Organization recommended diagnostics for TB reduced delays in diagnosing and initiating TB treatment. Future studies evaluating performance and impact of diagnostics should consider reporting time delay estimates based on the standardized reporting framework.