ADD a bunch of future work fluff

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Nathan Dwarshuis 2021-08-03 13:57:51 -04:00
parent c1ff03d5f7
commit f574b1b331
2 changed files with 59 additions and 8 deletions

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@ -2583,6 +2583,19 @@ CONCLUSIONS: We developed a simplified, semi-closed system for the initial selec
publisher = {The American Association of Immunologists},
}
@Article{Stephan2014,
author = {Sirkka B Stephan and Alexandria M Taber and Ilona Jileaeva and Ericka P Pegues and Charles L Sentman and Matthias T Stephan},
journal = {Nature Biotechnology},
title = {Biopolymer implants enhance the efficacy of adoptive T-cell therapy},
year = {2014},
month = {dec},
number = {1},
pages = {97--101},
volume = {33},
doi = {10.1038/nbt.3104},
publisher = {Springer Science and Business Media {LLC}},
}
@Comment{jabref-meta: databaseType:bibtex;}
@Comment{jabref-meta: grouping:

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@ -163,6 +163,7 @@
\newacronym{qbd}{QbD}{quality-by-design}
\newacronym{aws}{AWS}{amazon web services}
\newacronym{qpcr}{qPCR}{quantitative polymerase chain reaction}
\newacronym{cstr}{CSTR}{continuously stirred tank bioreactor}
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%
% SI units for uber nerds
@ -733,7 +734,7 @@ much higher surface area than a traditional flask when matched for volume.
Consequently, this means that microcarrier-based cultures can operate with much
lower footprints than flask-like systems. Microcarriers also allow cell culture
to operate more like a traditional chemical engineering process, wherein a
stirred tank bioreactor may be employed to enhance oxygen transfer, maintain pH,
\gls{cstr} may be employed to enhance oxygen transfer, maintain pH,
and continuously supply nutrients\cite{Derakhti2019}.
A variety of microcarriers have been designed, primarily differing in their
@ -4315,7 +4316,8 @@ group.
\section{future directions}
There are several important next steps to perform with this work:
There are several important next steps to perform with this work, many of which
will be relevent to using this technology in a clinical trial:
\subsection{Translation to GMP process}
@ -4341,12 +4343,33 @@ as dynabeads and thus the research-grade proteins used here could be easily
replaced. The \gls{snb} is a synthetic small molecule and thus does not have any
animal-origin concerns.
\subsection{Mechanistic investigation}
\subsection{mechanistic investigation}
% why do the dms work?
% can we put anything on the dms to enhance their potency?
Despite the improved outcomes in terms of expansion and phenotype relative to
beads, we don't have a good understanding of why they \gls{dms} platform works
as well as it does. Several broad areas remain to be investigated, including the
role of the increased cytokine output (including \il{15} which was explored to
some extent in this work), the role of cells on the interior of the \gls{dms}
relative to those outside the \gls{dms}, and the role of the physical surface
properties of the \gls{dms} (including the morphology and the stiffness).
\subsection{Assessing performance using unhealthy donors}
\subsection{additional ligands and signals on the DMSs}
In this work we only explored the use of \acd{3} and \acd{28} \glspl{mab} coated
on the surface of the \gls{dms}. The chemistry used for the \gls{dms} is very
general, and any molecule or protein that could be engineered with a biotin
ligand could be attached without any further modification. There are many other
ligands that could have profound effects on the expansion and quality of T cells
which may be utilized. The simplest next step is to simply vary the ratio of
\acd{3} and \acd{28} signal. Another obvious example is to attach
\il{15}/\il{15R$\upalpha$} complexes to the surface to mimic \textit{trans}
presentation from other cell types\cite{Stonier2010}. Other adhesion ligands or
peptides such as GFOGER could be used to stimulate T cells and provide more
motility on the \glspl{dms}\cite{Stephan2014}. Finally, viral delivery systems
could theoretically be attached to the \gls{dms}, greatly simplifying the
transduction step.
\subsection{assessing performance using unhealthy donors}
All the work presented in this dissertation was performed using healthy donors.
This was mostly due to the fact that it was much easier to obtain healthy donor
@ -4360,8 +4383,23 @@ expansion technology given that even in healthy donors, we observed the
\subsection{translation to bioreactors}
% use dms in non-static bioreactors such as wave by first activating in a static
% environment
In this work we performed some preliminary experiments demonstrating that the
\gls{dms} platform can work in a Grex bioreactor. While an important first step,
more work needs to be done to optimize how this system will or can work in a
scalable environment using bioreactors. There are several paths to explore.
Firstly, the Grex itself has additional automation accessories which could be
tested, which would allow continuous media exchange and cytokine
administration. While this is an improvement from the work done here, it is
still a Grex and has all the disadvantages of an open system. Secondly, other
static bioreactors such as the Quantum hollow fiber bioreactor (Terumo) could be
explored. Essentially the \gls{dms} would be an additional matrix that could be
supplied to this system which would enhance its compatibility with T cells.
Finally, suspension bioreactors such as the classic \gls{cstr} or WAVE
bioreactors could be tried. The caveat with these is that the T cells only seem
to be loosely attached to the \gls{dms} throughout culture, so an initial
activation/transduction step in static culture might be necessary before moving
to a suspension system (alternatively the \gls{dms} could be coated with
additional adhesion ligands to make the T cells attach more strongly).
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