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.

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.