Phase I clinical trial of autologous NK cell therapy using novel expansion method in patients with advanced digestive cancer

Table S2. Change in NK population in PBL

A multi-enzymatic cascade reaction for the stereoselective production of γ\gamma-oxyfunctionalyzed amino acids

A stereoselective three-enzyme cascade for synthesis of diasteromerically pure $\gamma$-oxyfunctionalized $\alpha$-amino acids was developed. By coupling a dynamic kinetic resolution (DKR) using an $\it N$-acylamino acid racemase (NAAAR) and an L-selective aminoacylase from $\textit {Geobacillus thermoglucosidasius}$ with a stereoselective isoleucine dioxygenase from $\textit {Bacillus thuringiensis,}$ diastereomerically pure oxidized amino acids were produced from racemic $\it N$-acetylamino acids. The three enzymes differed in their optimal temperature and pH-spectra. Their different metal cofactor dependencies led to inhibitory effects. Under optimized conditions, racemic $\it N$-acetylmethionine was quantitatively converted into L-methionine-$\textit {(S)}$-sulfoxide with 97% yield and 95% $\textit {de.}$ The combination of these three different biocatalysts allowed the direct synthesis of diastereopure oxyfunctionalized amino acids from inexpensive racemic starting material

Improvement of the process stability of arylmalonate decarboxylase by immobilization for biocatalytic profen synthesis

The enzyme arylmalonate decarboxylase (AMDase) enables the selective synthesis of enantiopure (S)-arylpropinates in a simple single-step decarboxylation of dicarboxylic acid precursors. However, the poor enzyme stability with a half-life time of about 1.2 h under process conditions is a serious limitation of the productivity, which results in a need for high catalyst loads. By immobilization on an amino C2 acrylate carrier the operational stability of the (S)-selective AMDase variant G74C/M159L/C188G/V43I/A125P/V156L was increased to a half-life of about 8.6 days, which represents a 158-fold improvement. Further optimization was achieved by simple immobilization of the cell lysate to eliminate the cost- and time intensive enzyme purification step

A combined experimental and modelling approach for the Weimberg pathway optimisation

The oxidative Weimberg pathway for the five-step pentose degradation to α-ketoglutarate is a key route for sustainable bioconversion of lignocellulosic biomass to added-value products and biofuels. The oxidative pathway from Caulobacter crescentus has been employed in in-vivo metabolic engineering with intact cells and in in-vitro enzyme cascades. The performance of such engineering approaches is often hampered by systems complexity, caused by non-linear kinetics and allosteric regulatory mechanisms. Here we report an iterative approach to construct and validate a quantitative model for the Weimberg pathway. Two sensitive points in pathway performance have been identified as follows: (1) product inhibition of the dehydrogenases (particularly in the absence of an efficient NAD+ recycling mechanism) and (2) balancing the activities of the dehydratases. The resulting model is utilized to design enzyme cascades for optimized conversion and to analyse pathway performance in C. cresensus cell-free extracts

Additional file 1: of Phase I clinical trial of autologous NK cell therapy using novel expansion method in patients with advanced digestive cancer

Table S1. Summary of NK cell expansion from 31 cancer patients (supplementary information of reference #25)

Phase I Clinical Trial of Fibronectin CH296-Stimulated T Cell Therapy in Patients with Advanced Cancer

<div><p>Background</p><p>Previous studies have demonstrated that less-differentiated T cells are ideal for adoptive T cell transfer therapy (ACT) and that fibronectin CH296 (FN-CH296) together with anti-CD3 resulted in cultured cells that contain higher amounts of less-differentiated T cells. In this phase I clinical trial, we build on these prior results by assessing the safety and efficacy of FN-CH296 stimulated T cell therapy in patients with advanced cancer.</p><p>Methods</p><p>Patients underwent fibronectin CH296-stimulated T cell therapy up to six times every two weeks and the safety and antitumor activity of the ACT were assessed. In order to determine immune function, whole blood cytokine levels and the number of peripheral regulatory T cells were analyzed prior to ACT and during the follow up.</p><p>Results</p><p>Transferred cells contained numerous less-differentiated T cells greatly represented by CD27+CD45RA+ or CD28+CD45RA+ cell, which accounted for approximately 65% and 70% of the total, respectively. No ACT related severe or unexpected toxicities were observed. The response rate among patients was 22.2% and the disease control rate was 66.7%.</p><p>Conclusions</p><p>The results obtained in this phase I trial, indicate that FN-CH296 stimulated T cell therapy was very well tolerated with a level of efficacy that is quite promising. We also surmise that expanding T cell using CH296 is a method that can be applied to other T- cell-based therapies.</p><p>Trial Registration</p><p>UMIN <a href="https://upload.umin.ac.jp/cgi-open-bin/ctr/ctr.cgi?function=brows&action=brows&type=summary&recptno=R000002101&language=J" target="_blank">UMIN000001835</a></p></div

Tumor response.

<p>CR = complete response; PR = partial response; SD = stable disease; PD = progressive disease; 95% CI = 95% confidence interval.</p

Characteristics of infused cells.

<p>Patient no. 1 to 6 needed a one-time culture and patients 7 to 9 underewent three lymphocyte cultures.</p><p>Values are expressed as mean±SD.</p