CAR T Cell Therapy: Fifteen years of academic driving

We have developed platforms to manufacture T lymphocytes expressing specific chimeric antigen receptors (CARs) that have enabled the successful implementation of multiple phase I/II CAR T cell clinical trials at MSKCC -including 2 multi center trials-. Subjects with leukemia, lymphoma, myeloma, breast cancer, mesothelioma, ovarian cancer and prostate cancer have been enrolled. Over 300 CAR T cell products have been successfully manufactured and more than 200 subjects have been infused across 13 phase I/II clinical trials. In order to support these trials, we established early on a robust platform using magnetic beads coated with agonistic anti-CD3 and anti-CD28 antibodies for the selection and activation of T cells, and the Wave/Xuri bioreactor for CAR T cell expansion. We also established our own process to manufacture replication-defective gammaretroviral vectors encoding CARs. This manufacturing platform consistently allows the generation of clinical doses in less than two weeks. Using this platform, the US Food and Drug Administration granted MSK Breakthrough Therapy Designation and Orphan Drug Designation in late 2014, for its CD19-targeted CAR therapy in patients with relapsed or refractory acute lymphoblastic leukemia yielding more than 85% complete remission. This vast experience provides many insights into addressing the substantial challenges that still remain to be resolved in order to broaden the usage of CAR T cells at the Point of Care and enable the commercialization of this therapeutic modality

Methods and Tools for decentralized on farm breeding

This technical booklet describes the possible experimental designs and statistical methods of analysis that can be carried out, according to the objectives and the experimental constraints of the breeding program nd the farmers’ group. The way to identify and select the most relevant devices and methods is based on a decision tree

Methods and tools for decentralized on farm breeding

This technical booklet describes the possible experimental designs and statistical methods of analysis that can be carried out, according to the objectives and the experimental constraints of the breeding program and the farmers’ group. The way to identify and select the most relevant devices and methods is based on a decision tree

Evaluation Of Population-Varieties Developed Within A Wheat Participatory Breeding Program In France: Performances, Diversity, Stability And Adaptation

Modern agricultural systems rely on little crop genetic diversity, especially with the use of homogeneous varieties grown on large areas. However crop genetic diversity within fields is a lever for a more sustainable production, allowing for a greater stability through combined resistances to biotic and abiotic stress, and buffering environmental heterogeneity which characterizes organic systems. In France, a Participatory Plant Breeding (PPB) project has been applied on bread wheat since 2006 involving farmers and facilitators of the farmers' seed network Réseau Semences Paysannes and INRA researchers for the development of populations based on a decentralized selection in farmers' fields. This project leads to the development of heterogeneous populations whose intra-variety genetic diversity should allow them to adapt to farmers' practices and environments. We evaluated the agronomic behavior, genetic diversity, stability and local adaptation of ten populations developed within the PPB program compared to two commercial pure line varieties. Some populations had very interesting responses when considering grain yield, biomass production and protein content, and six of them were not significantly less productive than the two commercial varieties when comparing overall grain yield per population. While no clear evidence of local adaptation was detected, we found that populations' quality and in a lesser extent grain yield were more stable over years than that of commercial varieties. Protein content stability over time was positively correlated to genetic diversity with no significant drawback on protein production

Advanced backcross QTL analysis and comparative mapping with RIL QTL studies and GWAS provide an overview of QTL and marker haplotype diversity for resistance to Aphanomyces root rot in pea (Pisum sativum)

Aphanomyces euteiches is the most damaging soilborne pea pathogen in France. Breeding of pea resistant varieties combining a diversity of quantitative trait loci (QTL) is a promising strategy considering previous research achievements in dissecting polygenic resistance to A. euteiches. The objective of this study was to provide an overview of the diversity of QTL and marker haplotypes for resistance to A. euteiches, by integrating a novel QTL mapping study in advanced backcross (AB) populations with previous QTL analyses and genome-wide association study (GWAS) using common markers. QTL analysis was performed in two AB populations derived from the cross between the susceptible spring pea variety “Eden” and the two new sources of partial resistance “E11” and “LISA”. The two AB populations were genotyped using 993 and 478 single nucleotide polymorphism (SNP) markers, respectively, and phenotyped for resistance to A. euteiches in controlled conditions and in infested fields at two locations. GWAS and QTL mapping previously reported in the pea-Aphanomyces collection and from four recombinant inbred line (RIL) populations, respectively, were updated using a total of 1,850 additional markers, including the markers used in the Eden x E11 and Eden x LISA populations analysis. A total of 29 resistance-associated SNPs and 171 resistance QTL were identified by GWAS and RIL or AB QTL analyses, respectively, which highlighted 10 consistent genetic regions confirming the previously reported QTL. No new consistent resistance QTL was detected from both Eden x E11 and Eden x LISA AB populations. However, a high diversity of resistance haplotypes was identified at 11 linkage disequilibrium (LD) blocks underlying consistent genetic regions, especially in 14 new sources of resistance from the pea-Aphanomyces collection. An accumulation of favorable haplotypes at these 11 blocks was confirmed in the most resistant pea lines of the collection. This study provides new SNP markers and rare haplotypes associated with the diversity of Aphanomyces root rot resistance QTL investigated, which will be useful for QTL pyramiding strategies to increase resistance levels in future pea varieties

Replication competent retrovirus testing (RCR) in the National Gene Vector Biorepository: No evidence of RCR in 1,595 post-treatment peripheral blood samples obtained from 60 clinical trials

The clinical impact of any therapy requires the product be safe and effective. Gammaretroviral vectors pose several unique risks, including inadvertent exposure to replication competent retrovirus (RCR) that can arise during vector manufacture. The US FDA has required patient monitoring for RCR, and the National Gene Vector Biorepository is an NIH resource that has assisted eligible investigators in meeting this requirement. To date, we have found no evidence of RCR in 338 pre-treatment and 1,595 post-treatment blood samples from 737 patients associated with 60 clinical trials. Most samples (75%) were obtained within 1 year of treatment, and samples as far out as 9 years after treatment were analyzed. The majority of trials (93%) were cancer immunotherapy, and 90% of the trials used vector products produced with the PG13 packaging cell line. The data presented here provide further evidence that current manufacturing methods generate RCR-free products and support the overall safety profile of retroviral gene therapy

The Warburg Effect Is Genetically Determined in Inherited Pheochromocytomas

The Warburg effect describes how cancer cells down-regulate their aerobic respiration and preferentially use glycolysis to generate energy. To evaluate the link between hypoxia and Warburg effect, we studied mitochondrial electron transport, angiogenesis and glycolysis in pheochromocytomas induced by germ-line mutations in VHL, RET, NF1 and SDH genes. SDH and VHL gene mutations have been shown to lead to the activation of hypoxic response, even in normoxic conditions, a process now referred to as pseudohypoxia. We observed a decrease in electron transport protein expression and activity, associated with increased angiogenesis in SDH- and VHL-related, pseudohypoxic tumors, while stimulation of glycolysis was solely observed in VHL tumors. Moreover, microarray analyses revealed that expression of genes involved in these metabolic pathways is an efficient tool for classification of pheochromocytomas in accordance with the predisposition gene mutated. Our data suggest an unexpected association between pseudohypoxia and loss of p53, which leads to a distinct Warburg effect in VHL-related pheochromocytomas