ADD conclusion paragraphs

tex/thesis.tex

@@ -3403,7 +3403,87 @@ this may explain the modest advantage that the \gls{dms} T cells seemed to have
 in the second experiment in slowing the progression of tumor burden.
 
 \chapter{conclusions and future work}\label{conclusions}
+
 \section{conclusions}
+
+This dissertation describes the development of a novel T cell expansion
+platform, including the fabrication, quality control, and biological validation
+of its performance both \invitro{} and \invivo{}. Development of such a system
+would be meaningful even if it only performed as well as current methods, as
+adding another method to the arsenal of the growing T cell manufacturing
+industry would reduce the reliance on a small number of companies that currently
+license magnetic bead-based T cell expansion technology. However, we
+additionally show that the \gls{dms} platform expands more T cells on average,
+including highly potent \ptmem{} and \pth{} T cells, and produces higher
+percentages of both. If commercialized, this would be a compelling asset the T
+cell manufacturing industry.
+
+% TODO double check the numbers at the end
+In \cref{aim1}, we develop the \gls{dms} platform and verified its efficacy
+\invitro{}. Importantly, this included \gls{qc} steps at every critical step of
+the fabrication process to ensure that the \gls{dms} can be made within a
+targeted specification. These \gls{qc} steps all rely on common, relatively
+cost-effective assays such as the \gls{haba} assay, \gls{bca} assay, and
+\glspl{elisa}, thus other labs and commercial entities should be able to perform
+them. The microcarriers themselves are an off-the-shelf product available from
+reputable vendors, further enhancing translatability. On average, we
+demonstrated that the \gls{dms} outperforms state-of-the-art bead-based T cell
+expansion technology in terms of total fold expansion, \ptmemp{}, and \pthp{} by
+\SI{143}{\percent}, \SI{2.5}{\percent}, and \SI{9.8}{\percent} controlling for
+donor, operator, and a variety of process conditions.
+
+In \cref{aim2a}, we developed a modeling pipeline that can be used by commercial
+entities as the scale up this process to identify \glspl{cqa} and \gls{cpp}.
+These are highly important for a variety of reasons. First, understanding
+pertinent \glspl{cpp} allow manufacturers to operate their process at optimal
+conditions. This is important for anti-tumor cell therapies, where the prospects
+of a patient can urgently depend on receiving therapy in a timely manner.
+Optimal process conditions allow T cells to be expanded as quickly as possible
+for the patient, while also minimizing cost for the manufacturer. Second,
+\glspl{cqa} can be used to define process control schemes as well as release
+criteria. Process control, and with it the ability to predict future outcomes
+based on data obtained at the present, is highly important for cell therapies
+given that batch failures are extremely expensive {\#}, and predicting a batch
+failure would allow manufacturers to restart the batch in a timely manner
+without wasting resources. Furthermore, \glspl{cqa} can be used to define what a
+`good' vs `bad' product is, which will important help anticipate dosing and
+followup procedures in the clinic if the T cells are administered. In the aim,
+we cannot claim to have found the ultimate set of \glspl{cqa} and \glspl{cpp},
+as we used tissue culture plates instead of a bioreactor and we only used one
+donor. However, we have indeed outlined a process that others may use to find
+these for their process. In particular, the 2-phase modeling process we used
+(starting with a \gls{doe} and collecting data longitudinally) is a strategy
+that manufacturers can easily implement. Also, collecting secretome and
+metabolome is easily generalized to any setting and to most bioreactors and
+expansion systems, as they can be obtained with relatively inexpensive equipment
+(Luminex assay, benchtop \gls{nmr}, etc) without disturbing the cell culture.
+
+In \cref{aim2b}, we further explored additional tuning knobs that could be used
+to control and optimize the \gls{dms} system. We determined that altering the
+\gls{dms} concentration temporally has profound effects on the phenotype and
+expansion rate. This agrees with other data we obtained in \cref{aim2a} and with
+what others have generally reported about signal strength and T cell
+differentiation {\#}. We did not find any mechanistic relationship between
+either integrin signaling or \gls{il15} signaling. In the case of the former, it
+may be more likely that the \glspl{dms} surfaces are saturated to the point of
+sterically hindering any integrin interactions with the collagen surface. In the
+case of \gls{il15} more experiments likely need to be done in order to plausibly
+rule out this mechanism and/or determine if it is involved at all.
+
+% TODO make this tighter and cite paper showing that this makes at least some
+% sense
+In \cref{aim3} we determined that the \glspl{dms} expand T cells that also
+performed better than beads \invivo{}. In the first experiment we performed, the
+results were very clearly in favor of the \glspl{dms}. In the second experiment,
+even the \gls{dms} group failed to fully control the tumor burden, but this is
+not surprising given the low \ptcarp{} across all groups. Also, despite this, the
+\gls{dms} group appeared to control the tumor better on average for early, mid,
+and late T cell harvesting timepoints. It was not clear if this effect was due
+to increased \cdp{} or overall increased fitness of the \gls{dms}-expanded T
+cells given their higher expansion rate. The \ptmemp{} did not seem to be a
+factor given that it was nearly the same in the first experiment between
+\gls{dms} and bead groups despite the clear advantage seen in the \gls{dms} group.
+
 \section{future work}
 
 \onecolumn