diff --git a/tex/thesis.tex b/tex/thesis.tex index f101b0e..dd9eaf9 100644 --- a/tex/thesis.tex +++ b/tex/thesis.tex @@ -2024,6 +2024,22 @@ likely serve as a drop-in substitution to provide these benefits. \chapter{aim 2a}\label{aim2a} \section{introduction} + +The purpose of this sub-aim was to develop computational methods to identify novel +\glspl{cqa} and \glspl{cpp} that could be used for release criteria, process +control, and process optimization for the \gls{dms} platform. We hypothesized +that T cells grown using the \gls{dms} system would produce detectable +biological signatures in the media supernatent which corresponded to clinically +relevent responses such as the fold expansion of the T cells or the resulting +phenotype. We tested this hypothesis by activating T cells under a variety of +conditions using a \gls{doe}, sampling the media at intermediate timepoints, and +creating models to predict the outcome of the cultures. We should stress that +the specific \glspl{cpp} and \glspl{cqa} determined by this aim are not +necessarily universal, as this was not performed with equipment that would +normally be used at scale. However, the process outlined here is one that can +easily be adaptable to any system, and the specific findings themselves offer +interesting insights that warrant further study. + \section{methods} \subsection{study design} @@ -2776,6 +2792,15 @@ sensors and controls for formate, lactate, along with ethanol and glucose. \chapter{aim 2b}\label{aim2b} \section{introduction} + +The purpose of this sub-aim was to further explore the \gls{dms} platform, +specifically for mechanisms and pathways that could be the basis for additional +\glspl{cpp} that could be optimized to yield higher quantity and quality of T +cells. Our strategy in general was to perturb the \gls{dms} system from the +normal operating conditions at which it was used up until this point either +through temporal modulation of activation signal or by blocking pathways of +interest using \glspl{mab}. + \section{methods} \subsection{DMSs temporal modulation} @@ -3246,6 +3271,21 @@ advantage via \gls{il15} signaling. \chapter{aim 3}\label{aim3} \section{introduction} + +% DO NOT COMMENT OUT THIS LINE: the real purpose of this aim was to appease +% Nature Biotech because they think that animal models are necessary for good +% science. This entire aim exists because of their foolishness. + +The purpose of this aim was to verify that \gls{car} T cells produced using the +\gls{dms} system will show potent anti-tumor properties in a complex \invivo{} +system compared to state-of-the-art bead technology. We hypothesized that due to +the increased \ptmem{} and \pth{} phenotypes as shown in \cref{aim1}, that +\gls{dms}-expanded T cells would show longer survival and lower tumor burden +than those expanded with beads. We explored the effect of dosing at different +levels and the effect of harvesting T cells at early timepoints in the culture, +which has been shown to produce lower-differentiated T cells with higher +potency\cite{Ghassemi2018}. + \section{methods} \subsection{CD19-CAR T cell generation}