Chimeric antigen receptor T-cells: Properties, production, and quality control.

Chimeric antigen receptor (CAR) T-cell therapy is a novel adoptive T-cell immunotherapy for haematological malignancies. First introduced into clinical practice in 2017, CAR T-cell therapy is now finding its place in the management of lymphoid malignancies, primarily of B-cell lineage, including lymphoblastic leukaemia, non-Hodgkin lymphoma and plasma cell myeloma, with remarkable therapeutic outcomes. CAR T-cells are a customised therapeutic product for each patient. Manufacture commences with collection of autologous T-cells, which are then genetically engineered ex vivo to express transmembrane CARs. These chimeric proteins consist of an antibody-like extracellular antigen-binding domain, to recognise specific antigens on the surface of tumour cells (e.g. CD19), linked to the intracellular co-stimulatory signalling domains of a T-cell receptor (e.g. CD137). The latter is required for in vivo CAR T-cell proliferation, survival, and durable efficacy. Following reinfusion, CAR T-cells harness the cytotoxic capacity of a patient's immune system. They overcome major mechanisms of tumour immuno-evasion and have potential to generate robust cytotoxic anti-tumour responses. This review discusses the background to CAR T-cell therapies, including their molecular design, mechanisms of action, methods of production, clinical applications and established and emerging technologies for CAR T-cell evaluation. It highlights the need for standardisation, quality control and monitoring of CAR T-cell therapies, to ensure their safety and efficacy in clinical management.

Is Maternal Serum Homocysteine a Novel Diagnostic Biomarker for Predicting Placenta-Mediated Disorders?

Background Uteroplacental insufficiency and related disorders, though a significant cause of undesirable maternal and fetal outcomes, are complex and poorly understood. The newer screening modalities are expensive and difficult to procure for day-to-day use in developing countries. This study aimed to examine the association of mid-trimester maternal serum homocysteine levels with maternal and neonatal outcomes. Methodology This was a prospective cohort study involving 100 participants between 18 and 28 weeks of gestation. The study was conducted at a tertiary care center in south India from July 2019 to September 2020. Maternal blood samples were analyzed for serum homocysteine levels and correlated with the third-trimester pregnancy outcomes. Statistical analysis was done, and diagnostic measures were computed. Results The mean age was found to be 26.8 ± 4.8 years. Of the participants, 15% (n = 15) were diagnosed with hypertensive disorders during pregnancy, while 7% (n = 7) had fetal growth restriction (FGR) and 7% (n = 7) were complicated by preterm birth. An elevated maternal serum homocysteine level was positively associated with adverse pregnancy outcome measures such as hypertensive disorders (p = 0.001), with sensitivity and specificity of 27% and 99%, respectively, and FGR (p = 0.03) with sensitivity and specificity of 28.6% and 98.6%, respectively. Moreover, a statistically significant outcome was noted with preterm birth before 37 weeks (p = 0.001) and a low Apgar score (p = 0.02). No association was established with spontaneous preterm labor (p = 1.00), neonatal birth weight (p = 0.42), and special care unit admission (p = 1.00). Conclusions Such a simple and affordable investigation has the potential to go a long way in the early diagnosis and management of placenta-mediated disorders in pregnancy during the antenatal period, especially in low-resource settings.

Role of Routine Mid-Trimester Uterine Artery Doppler for Surveillance of Placental Mediated Disorders in a Low-Risk Population.

OBJECTIVES: Abnormalities in the placentation process can increase pregnancy-related complications like pre-eclampsia, placental abruption, intrauterine-fetal death (IUFD) or foetal-growth restriction (FGR). Our objective was to investigate the feasibility of utilising the mid-trimester uterine artery Doppler Pulsatility Index (PI), a non-invasive and effective screening tool, as a diagnostic measure to predict adverse pregnancy outcomes in a low-risk population in South India. MATERIALS AND METHODS: This prospective cohort study was done in the Obstetrics and Gynaecology unit at Amrita Institute of Medical Sciences, South India, between August 2018 and January 2020. Uterine artery Doppler was performed along with the targeted anomaly scan between 18 and 24 weeks of gestation and a relationship was established with pregnancy outcome. RESULTS: Of 100 participants, abnormal uterine artery PI (PI > 90th centile) was found in 13 pregnancies, of which statistically significant association was found with hypertensive disorders (P=0.001), FGR (P=0.064) and preterm birth before 37 weeks (P=0.051). No association was found between abnormal uterine artery PI and neonatal birth weight (P=-0.3), APGAR score (P=0.35) and NICU admission (P=0.078). CONCLUSION: An early abnormal finding in the doppler study can modify the level of antenatal surveillance required along with appropriate timely interventions, thereby significantly reducing the associated maternal and neonatal morbidity and mortality. When combined with routine ultrasound in pregnancy, such an affordable and straightforward diagnostic modality can improve antenatal care by reducing complications even in a low-risk population.

Association of combined second trimester maternal serum Homocysteine and Uterine Artery Doppler to predict adverse pregnancy outcome.

Introduction: Disturbances in placentation increase the risk of maternal and fetal complications. Several biochemical and imaging modalities have been studied, but the hunt for a single effective screening test never became a reality as the causes of this complex condition are multifactorial and polygenetic, many of which we are only beginning to discover. Not many studies have been conducted in the developing countries like India and other low resource settings to consider whether it would be worthwhile to combine inexpensive and effective markers together for better prediction of adverse pregnancy outcome.This study primarily aims to investigate the predictability of combined screening with maternal serum homocysteine and second trimester uterine artery Doppler in diagnosis of adverse pregnancy outcome. Methodology: A prospective cohort study which involved 100 women with singleton gestation, meeting the inclusion criteria, attending the inpatient or outpatient of Obstetrics and Gynaecology in Amrita Institute of Medical Sciences, Kerala, a tertiary care centre in Southern India from July 2016 and September 2018 was conducted. Serum Homocysteine estimation (tHcy) was done between 18 and 28 weeks of gestation with informed consent, and uterine artery (UA) Doppler PI which is a non-invasive routine study was done along with targeted second trimester anomaly scan (18-24 weeks) in Fetal Medicine Department. Cutoff values of tHcy and UA PI were computed at 95th (> / = 9.7 mmol/l) and 90th percentile, respectively as reported by Onalan et al. [9] and Nicholaides et al. [4]. Statistical analysis was performed using IBM SPSS version 20.0 software. Chi-square test and diagnostic measures were also used. Results: Of the 100 patients, 15% (n = 15) developed hypertensive disorder. 7% (n = 7) had FGR and 7%(n = 7) had spontaneous preterm birth. 6% (n = 6) neonates had an APGAR score < 7 and 8% neonates (n = 8) required immediate NICU admission. Statistically significant association was found when tHcy and UA PI were used together for the prediction of FGR (p = 0.003), preterm birth (p = 0.002) and low APGAR score at birth (p = 0.009) with a specificity of 83.4%. With regard to PIH, both parameters were found to be statistically significant only when used independently (p = 0.001) but not when used in combination (p = 0.17). Both elevated tHcy and abnormal UA PI used in combination predicted adverse pregnancy outcome like FGR but with a low sensitivity of 14.3% and high specificity of 98.9%. However, when used independently these markers predicted FGR with a better sensitivity (tHcy- 28.6% and UA PI- 44.4%). Conclusion: Findings from this study have been promising with potential clinical implications for the diagnosis and management of high-risk pregnancies. Though the independent role of the two markers in screening various adverse pregnancy outcomes could be proved, their combined use to improve predictivity of more complications warrants further studies on a larger population with appropriate randomisation.

Knowledge, attitude and practice on usage and toxicity of local anaesthetic agents (LA) amongst health care professionals in obstetrics and gynaecology.

A sound knowledge of LA and their dosage is essential for effective anaesthesia and patient safety as LA toxicity could be potentially life-threatening and clinicians undertaking LA procedures must be competent in early recognition of toxicity and instigating remedial measures timely. This is a prospective national-level survey using self-administered questionnaires among health care professionals in Obstetrics and Gynaecology primarily based in Worcestershire Acute Hospital NHS Trust and further extended to other regions of the United Kingdom. The survey was focussed on evaluating the knowledge, attitude and practice (KAP) of the common local anaesthetic agents, their dosage, early recognition and management of LA systemic toxicity(LAST). We understood that all groups of health care professionals did not demonstrate adequate knowledge of safe and effective use of LA agents. We thereby propose an accredited knowledge-based learning module, laminated safety information cards for LA use and ensuring availability of Lipid-emulsion therapy in clinical areas.IMPACT STATEMENTWhat is already known on this subject? Health care professionals using LA should have sufficient knowledge of LA including the required and maximum dose allowance, pharmacokinetic properties, possible complications and management of systemic toxicity. The lack this can trigger potentially fatal and life-threatening complications if misused. Therefore, the key objectives of the use of LA in clinical settings are to optimise pain management while ensuring safe practice.What do the results of this study add? We understood that all groups of health care professionals did not demonstrate adequate knowledge of the safe and effective use of LA agents and appropriate management of its toxicity. This highlights the urgent need for extending awareness among the professionals on the safe administration of LA, early identification and prompt management of LAST. This will improve not only the safety but also the overall patient experience and quality of care.What are the implications of these findings for clinical practice and/or further research? We thereby propose that an accredited knowledge-based module on safe administration and safety concerns of LA must be made compulsory for all those administering LA. Laminated safety information cards on LA with explicit algorithms/guidelines should be made available in all clinical areas to avoid any delays in the management of toxicity. Moreover, all health-care institutions must ensure the availability of Lipid Emulsion Therapy.

Amyloid beta acts synergistically as a pro-inflammatory cytokine.

The amyloid beta (Aß) peptide is believed to play a central role in Alzheimer's disease (AD), the most common age-related neurodegenerative disorder. However, the natural, evolutionarily selected functions of Aß are incompletely understood. Here, we report that nanomolar concentrations of Aß act synergistically with known cytokines to promote pro-inflammatory activation in primary human astrocytes (a cell type increasingly implicated in brain aging and AD). Using transcriptomics (RNA-seq), we show that Aß can directly substitute for the complement component C1q in a cytokine cocktail previously shown to induce astrocyte immune activation. Furthermore, we show that astrocytes synergistically activated by Aß have a transcriptional signature similar to neurotoxic "A1" astrocytes known to accumulate with age and in AD. Interestingly, we find that this biological action of Aß at low concentrations is distinct from the transcriptome changes induced by the high/supraphysiological doses of Aß often used in in vitro studies. Collectively, our results suggest an important, cytokine-like function for Aß and a novel mechanism by which it may directly contribute to the neuroinflammation associated with brain aging and AD.

Proteomic variations of esophageal squamous cell carcinoma revealed by combining RNA-seq proteogenomics and G-PTM search strategy.

BACKGROUND: Cancer that arises from epithelial cells of the esophagus is called esophagus squamous cell carcinoma (ESCC) and is mostly observed in developing nations. Evaluation of cancer genomes and its regulation into proteins plays a predominant role in understanding the cancer progressions. Mass-spectrometry-based proteomics is a consequential tool to estimate proteomic variation and posttranslational modifications (PTMs) from standard protein databases. Post-translational modifications play a crucial role in protein folding and PTMs can be accounted for as a biological signal to interpret the structural changes and transition order of proteins. Functional validation of cancer-related mutations can explain the effects of mutations on genes and the identification of Oncogenes and tumor suppressor genes. Therefore, we present a study on protein variations to interpret the structural changes and transition order of proteins in ESCC carcinogenesis. METHODOLOGY: We are using a bottom-up proteomics approach with Galaxy-P framework and RNA sequence data analysis to generate the sample-specific databases containing details of RNA splicing and variant peptides. Once the database generated with information on variable modification, only the curated PTMs at specific positions are considered to perform spectral matching. Proteogenomics mapping was performed to identify protein variations in ESCC. RESULTS: RNA-sequence proteogenomics with G-PTM (Global Post-Translational Modification) searching strategy has revealed proteomic events including several peptides that contain single amino acid variations, novel splice junction peptides and posttranslationally modified peptides. Proteogenomic mapping exhibited the splice junction peptides mapped predominantly for Malic enzyme exon type (ME-3) and MCM7 protein-coding genes that promote cancer progression, found to be exhibited in ESCC samples. Approximately 25 ± types of PTM modifications were recorded, and Protein Phosphorylation was largely noted. CONCLUSION: ESCC cancer prognosis at the molecular level enables a better understanding of cancer carcinogenesis and protein modifications can be used as potential biomarkers.

Pregnancy-Associated Breast Cancer: A Realistic Approach.

BACKGROUND: The number of cancers diagnosed during pregnancy is on the rise, and breast cancer is the most common malignancy. Presently, there are very limited resources and no clear guidelines for managing this peculiar patient population both worldwide and in India. The objective of this study was to find out the incidence of pregnancy-associated breast cancer (PABC) in a tertiary care referral centre and to compare the epidemiological, diagnostic and prognostic factors as well as maternal and foetal outcomes with the most recent literature worldwide. METHODS: We conducted a retrospective descriptive study of women diagnosed with breast cancer in pregnancy and post-partum period at a tertiary care centre in southern India during the period of 10 years (total number of breast cancer patients were 10). We studied the diagnostic and prognostic factors as well as maternal and foetal outcome in patients diagnosed with breast cancer for the first time in pregnancy. RESULTS: Overall incidence of PABC was found to be 0.6% (n = 10). Mean age at the time of presentation was 30.7 ± 4 years. All cases suspected clinically or on imaging (USG) were confirmed with FNAC, excision biopsy or Trucut biopsy. Out of 10 patients, 70% (n = 7) had an advanced-stage disease on diagnosis. Histopathology suggested 90% (n = 9) had invasive ductal carcinoma and 55.5% (n = 5) had a triple negative receptor status. 20% (n = 2) of our patients had opted for a breast conservation surgery (BCS), and 70% (n = 7) of our patients underwent modified radical mastectomy with neoadjuvant or adjuvant chemotherapy/radiotherapy. One patient had a second trimester MTP in view of stage 4 disease. 77.7% (n = 7) of the nine patients who continued pregnancy underwent LSCS, out of which 57.4% (n = 4) were elective, and MRM was done concurrently with LSCS in 50% (n = 2) of the elective LSCS. The mean birth weight of the 9 neonates was 2.2 ± 0.5 kg. Intrauterine growth retardation was seen in 22.2% (n = 2) neonates. 33.3% (n = 3) of the neonates required NICU support, and one baby expired on post-natal day 16. CONCLUSION: With the increasing number of elderly primigravida amongst the urban population, a clear understanding of PABC is becoming more important. A multidisciplinary team approach shall help the clinician not only in reducing the heavy burden of patient responsibility but more importantly, in guaranteeing better quality of treatment, avoiding unnecessary delays in providing interventions and providing adequate treatment.