diff --git a/tex/proposal.bib b/tex/references.bib similarity index 100% rename from tex/proposal.bib rename to tex/references.bib diff --git a/tex/thesis.bbl b/tex/thesis.bbl new file mode 100644 index 0000000..94d9ebb --- /dev/null +++ b/tex/thesis.bbl @@ -0,0 +1,210 @@ +\begin{thebibliography}{10} +\expandafter\ifx\csname url\endcsname\relax + \def\url#1{\texttt{#1}}\fi +\expandafter\ifx\csname urlprefix\endcsname\relax\def\urlprefix{URL }\fi +\providecommand{\bibinfo}[2]{#2} +\providecommand{\eprint}[2][]{\url{#2}} + +\bibitem{Fesnak2016} +\bibinfo{author}{Fesnak, A.~D.}, \bibinfo{author}{June, C.~H.} \& + \bibinfo{author}{Levine, B.~L.} +\newblock \bibinfo{title}{Engineered t cells: the promise and challenges of + cancer immunotherapy}. +\newblock \emph{\bibinfo{journal}{Nature Reviews Cancer}} + \textbf{\bibinfo{volume}{16}}, \bibinfo{pages}{566--581} + (\bibinfo{year}{2016}). + +\bibitem{Rosenberg2015} +\bibinfo{author}{Rosenberg, S.~A.} \& \bibinfo{author}{Restifo, N.~P.} +\newblock \bibinfo{title}{Adoptive cell transfer as personalized immunotherapy + for human cancer}. +\newblock \emph{\bibinfo{journal}{Science}} \textbf{\bibinfo{volume}{348}}, + \bibinfo{pages}{62--68} (\bibinfo{year}{2015}). + +\bibitem{Roddie2019} +\bibinfo{author}{Roddie, C.}, \bibinfo{author}{O'Reilly, M.}, + \bibinfo{author}{Pinto, J. D.~A.}, \bibinfo{author}{Vispute, K.} \& + \bibinfo{author}{Lowdell, M.} +\newblock \bibinfo{title}{Manufacturing chimeric antigen receptor t cells: + issues and challenges}. +\newblock \emph{\bibinfo{journal}{Cytotherapy}} (\bibinfo{year}{2019}). + +\bibitem{Dwarshuis2017} +\bibinfo{author}{Dwarshuis, N.~J.}, \bibinfo{author}{Parratt, K.}, + \bibinfo{author}{Santiago-Miranda, A.} \& \bibinfo{author}{Roy, K.} +\newblock \bibinfo{title}{Cells as advanced therapeutics: State-of-the-art, + challenges, and opportunities in large scale biomanufacturing of high-quality + cells for adoptive immunotherapies}. +\newblock \emph{\bibinfo{journal}{Advanced Drug Delivery Reviews}} + \textbf{\bibinfo{volume}{114}}, \bibinfo{pages}{222--239} + (\bibinfo{year}{2017}). + +\bibitem{Wang2016} +\bibinfo{author}{Wang, X.} \& \bibinfo{author}{Rivi{\`{e}}re, I.} +\newblock \bibinfo{title}{Clinical manufacturing of {CAR} t cells: foundation + of a promising therapy}. +\newblock \emph{\bibinfo{journal}{Molecular Therapy - Oncolytics}} + \textbf{\bibinfo{volume}{3}}, \bibinfo{pages}{16015} (\bibinfo{year}{2016}). + +\bibitem{Piscopo2017} +\bibinfo{author}{Piscopo, N.~J.} \emph{et~al.} +\newblock \bibinfo{title}{Bioengineering solutions for manufacturing challenges + in {CAR} t cells}. +\newblock \emph{\bibinfo{journal}{Biotechnology Journal}} + \textbf{\bibinfo{volume}{13}}, \bibinfo{pages}{1700095} + (\bibinfo{year}{2017}). + +\bibitem{Bashour2015} +\bibinfo{author}{Bashour, K.~T.} \emph{et~al.} +\newblock \bibinfo{title}{Functional characterization of a t cell stimulation + reagent for the production of therapeutic chimeric antigen receptor t cells}. +\newblock \emph{\bibinfo{journal}{Blood}} \textbf{\bibinfo{volume}{126}}, + \bibinfo{pages}{1901--1901} (\bibinfo{year}{2015}). + +\bibitem{Gendron2003} +\bibinfo{author}{Gendron, S.}, \bibinfo{author}{Couture, J.} \& + \bibinfo{author}{Aoudjit, F.} +\newblock \bibinfo{title}{Integrin $\alpha$2$\beta$1inhibits fas-mediated + apoptosis in t lymphocytes by protein phosphatase 2a-dependent activation of + the {MAPK}/{ERK} pathway}. +\newblock \emph{\bibinfo{journal}{Journal of Biological Chemistry}} + \textbf{\bibinfo{volume}{278}}, \bibinfo{pages}{48633--48643} + (\bibinfo{year}{2003}). + +\bibitem{Ohtani2008} +\bibinfo{author}{Ohtani, O.} \& \bibinfo{author}{Ohtani, Y.} +\newblock \bibinfo{title}{Structure and function of rat lymph nodes}. +\newblock \emph{\bibinfo{journal}{Archives of Histology and Cytology}} + \textbf{\bibinfo{volume}{71}}, \bibinfo{pages}{69--76} + (\bibinfo{year}{2008}). + +\bibitem{Boisvert2007} +\bibinfo{author}{Boisvert, M.}, \bibinfo{author}{Gendron, S.}, + \bibinfo{author}{Chetoui, N.} \& \bibinfo{author}{Aoudjit, F.} +\newblock \bibinfo{title}{Alpha2beta1 integrin signaling augments t cell + receptor-dependent production of interferon-gamma in human t cells}. +\newblock \emph{\bibinfo{journal}{Molecular Immunology}} + \textbf{\bibinfo{volume}{44}}, \bibinfo{pages}{3732--3740} + (\bibinfo{year}{2007}). + +\bibitem{Ben-Horin2004} +\bibinfo{author}{Ben-Horin, S.} \& \bibinfo{author}{Bank, I.} +\newblock \bibinfo{title}{The role of very late antigen-1 in immune-mediated + inflammation}. +\newblock \emph{\bibinfo{journal}{Clinical Immunology}} + \textbf{\bibinfo{volume}{113}}, \bibinfo{pages}{119--129} + (\bibinfo{year}{2004}). + +\bibitem{Forget2014} +\bibinfo{author}{Forget, M.-A.} \emph{et~al.} +\newblock \bibinfo{title}{Activation and propagation of tumor-infiltrating + lymphocytes on clinical-grade designer artificial antigen-presenting cells + for adoptive immunotherapy of melanoma}. +\newblock \emph{\bibinfo{journal}{Journal of Immunotherapy}} + \textbf{\bibinfo{volume}{37}}, \bibinfo{pages}{448--460} + (\bibinfo{year}{2014}). + +\bibitem{Cheung2018} +\bibinfo{author}{Cheung, A.~S.}, \bibinfo{author}{Zhang, D. K.~Y.}, + \bibinfo{author}{Koshy, S.~T.} \& \bibinfo{author}{Mooney, D.~J.} +\newblock \bibinfo{title}{Scaffolds that mimic antigen-presenting cells enable + ex vivo expansion of primary {T} cells}. +\newblock \emph{\bibinfo{journal}{Nature Biotechnology}} + \textbf{\bibinfo{volume}{36}}, \bibinfo{pages}{160--169} + (\bibinfo{year}{2018}). + +\bibitem{Rio2018} +\bibinfo{author}{del R{\'{\i}}o, E.~P.}, \bibinfo{author}{Miguel, M.~M.}, + \bibinfo{author}{Veciana, J.}, \bibinfo{author}{Ratera, I.} \& + \bibinfo{author}{Guasch, J.} +\newblock \bibinfo{title}{Artificial 3d culture systems for t cell expansion}. +\newblock \emph{\bibinfo{journal}{{ACS} Omega}} \textbf{\bibinfo{volume}{3}}, + \bibinfo{pages}{5273--5280} (\bibinfo{year}{2018}). + +\bibitem{Delalat2017} +\bibinfo{author}{Delalat, B.} \emph{et~al.} +\newblock \bibinfo{title}{{3D printed lattices as an activation and expansion + platform for T cell therapy}}. +\newblock \emph{\bibinfo{journal}{Biomaterials}} + \textbf{\bibinfo{volume}{140}}, \bibinfo{pages}{58--68} + (\bibinfo{year}{2017}). + +\bibitem{meyer15_immun} +\bibinfo{author}{Meyer, R.~A.} \emph{et~al.} +\newblock \bibinfo{title}{Immunoengineering: Biodegradable nanoellipsoidal + artificial antigen presenting cells for antigen specific t-cell activation + (small 13/2015)}. +\newblock \emph{\bibinfo{journal}{Small}} \textbf{\bibinfo{volume}{11}}, + \bibinfo{pages}{1612--1612} (\bibinfo{year}{2015}). + +\bibitem{Lambert2017} +\bibinfo{author}{Lambert, L.~H.} \emph{et~al.} +\newblock \bibinfo{title}{{Improving T Cell Expansion with a Soft Touch.}} +\newblock \emph{\bibinfo{journal}{Nano letters}} \textbf{\bibinfo{volume}{17}}, + \bibinfo{pages}{821--826} (\bibinfo{year}{2017}). + +\bibitem{Xu2014} +\bibinfo{author}{Xu, Y.} \emph{et~al.} +\newblock \bibinfo{title}{Closely related t-memory stem cells correlate with in + vivo expansion of car.cd19-t cells and are preserved by il-7 and il-15.} +\newblock \emph{\bibinfo{journal}{Blood}} \textbf{\bibinfo{volume}{123}}, + \bibinfo{pages}{3750--3759} (\bibinfo{year}{2014}). + +\bibitem{Fraietta2018} +\bibinfo{author}{Fraietta, J.~A.} \emph{et~al.} +\newblock \bibinfo{title}{Determinants of response and resistance to {CD}19 + chimeric antigen receptor ({CAR}) t cell therapy of chronic lymphocytic + leukemia}. +\newblock \emph{\bibinfo{journal}{Nature Medicine}} + \textbf{\bibinfo{volume}{24}}, \bibinfo{pages}{563--571} + (\bibinfo{year}{2018}). + +\bibitem{Gattinoni2011} +\bibinfo{author}{Gattinoni, L.} \emph{et~al.} +\newblock \bibinfo{title}{A human memory t cell subset with stem + cell{\textendash}like properties}. +\newblock \emph{\bibinfo{journal}{Nature Medicine}} + \textbf{\bibinfo{volume}{17}}, \bibinfo{pages}{1290--1297} + (\bibinfo{year}{2011}). + +\bibitem{Gattinoni2012} +\bibinfo{author}{Gattinoni, L.}, \bibinfo{author}{Klebanoff, C.~A.} \& + \bibinfo{author}{Restifo, N.~P.} +\newblock \bibinfo{title}{{Paths to stemness: building the ultimate antitumour + T cell.}} +\newblock \emph{\bibinfo{journal}{Nature reviews. Cancer}} + \textbf{\bibinfo{volume}{12}}, \bibinfo{pages}{671--84} + (\bibinfo{year}{2012}). + +\bibitem{Wang2018} +\bibinfo{author}{Wang, D.} \emph{et~al.} +\newblock \bibinfo{title}{Glioblastoma-targeted {CD}4+ {CAR} t cells mediate + superior antitumor activity}. +\newblock \emph{\bibinfo{journal}{{JCI} Insight}} \textbf{\bibinfo{volume}{3}} + (\bibinfo{year}{2018}). + +\bibitem{Yang2017} +\bibinfo{author}{Yang, Y.} \emph{et~al.} +\newblock \bibinfo{title}{{TCR} engagement negatively affects {CD}8 but not + {CD}4 {CAR} t cell expansion and leukemic clearance}. +\newblock \emph{\bibinfo{journal}{Science Translational Medicine}} + \textbf{\bibinfo{volume}{9}}, \bibinfo{pages}{eaag1209} + (\bibinfo{year}{2017}). + +\bibitem{Heathman2015} +\bibinfo{author}{Heathman, T. R.~J.} \emph{et~al.} +\newblock \bibinfo{title}{Expansion, harvest and cryopreservation of human + mesenchymal stem cells in a serum-free microcarrier process}. +\newblock \emph{\bibinfo{journal}{Biotechnology and Bioengineering}} + \textbf{\bibinfo{volume}{112}}, \bibinfo{pages}{1696--1707} + (\bibinfo{year}{2015}). + +\bibitem{Sart2011} +\bibinfo{author}{Sart, S.}, \bibinfo{author}{Errachid, A.}, + \bibinfo{author}{Schneider, Y.-J.} \& \bibinfo{author}{Agathos, S.~N.} +\newblock \bibinfo{title}{Controlled expansion and differentiation of + mesenchymal stem cells in a microcarrier based stirred bioreactor}. +\newblock \emph{\bibinfo{journal}{{BMC} Proceedings}} + \textbf{\bibinfo{volume}{5}} (\bibinfo{year}{2011}). + +\end{thebibliography} diff --git a/tex/thesis.tex b/tex/thesis.tex index 20255ec..6244ffc 100644 --- a/tex/thesis.tex +++ b/tex/thesis.tex @@ -48,6 +48,12 @@ \newacronym{cpp}{CPP}{critical process parameter} \newacronym{dms}{DMS}{degradable microscaffold} \newacronym{doe}{DOE}{design of experiments} +\newacronym{gmp}{GMP}{Good Manufacturing Practices} +\newacronym{cho}{CHO}{Chinese hamster ovary} +\newacronym{all}{ALL}{acute lymphoblastic leukemia} +\newacronym{pdms}{PDMS}{polydimethylsiloxane} +\newacronym{dc}{DC}{dendritic cell} +\newacronym{il2}{IL2}{interleukin 2} \newcommand{\mytitle}{ \Large{ @@ -234,6 +240,87 @@ quality in an industrial setting. \chapter{introduction} +T cell-based immunotherapies have received great interest from clinicians and +industry due to their potential to treat, and often cure, cancer and other +diseases\cite{Fesnak2016,Rosenberg2015}. In 2017, Novartis and Kite Pharma +received FDA approval for \textit{Kymriah} and \textit{Yescarta} respectively, +two genetically-modified \gls{car} T cell therapies against B cell malignancies. +Despite these successes, \gls{car} T cell therapies are constrained by an +expensive and difficult-to-scale manufacturing process with little control on +cell quality and phenotype3,4. State-of-the-art T cell manufacturing techniques +focus on anti-CD3 and anti-CD28 activation and expansion, typically presented on +superparamagnetic, iron-based microbeads (Invitrogen Dynabead, Miltenyi MACS +beads), on nanobeads (Miltenyi TransACT), or in soluble tetramers +(Expamer)\cite{Roddie2019,Dwarshuis2017,Wang2016, Piscopo2017, Bashour2015}. +These strategies overlook many of the signaling components present in the +secondary lymphoid organs where T cells expand in vivo. Typically, T cells are +activated under close cell-cell contact, which allows for efficient +autocrine/paracrine signaling via growth-stimulating cytokines such as +\gls{il2}. Additionally, the lymphoid tissues are comprised of \gls{ecm} +components such as collagen, which provide signals to upregulate proliferation, +cytokine production, and pro-survival pathways\cite{Gendron2003, Ohtani2008, + Boisvert2007, Ben-Horin2004}. We hypothesized that culture conditions that +better mimic these in vivo expansion conditions of T cells, can significantly +improve the quality and quantity of manufactured T cells and provide better +control on the resulting T cell phenotype. + +% TODO mention the Cloudz stuff that's in my presentation + +A variety of solutions have been proposed to make the T cell expansion process +more physiological. One strategy is to use modified feeder cell cultures to +provide activation signals similar to those of \glspl{dc}\cite{Forget2014}. +While this has the theoretical capacity to mimic many components of the lymph +node, it is hard to reproduce on a large scale due to the complexity and +inherent variability of using cell lines in a fully \gls{gmp}-compliant manner. +Others have proposed biomaterials-based solutions to circumvent this problem, +including lipid-coated microrods\cite{Cheung2018}, 3D-scaffolds via either +Matrigel\cite{Rio2018} or 3d-printed lattices\cite{Delalat2017}, ellipsoid +beads\cite{meyer15_immun}, and \gls{mab}-conjugated \gls{pdms} +beads\cite{Lambert2017} that respectively recapitulate the cellular membrane, +large interfacial contact area, 3D-structure, or soft surfaces T cells normally +experience in vivo. While these have been shown to provide superior expansion +compared to traditional microbeads, none of these methods has been able to show +preferential expansion of functional naïve/memory and CD4 T cell populations. +Generally, T cells with a lower differentiation state such as naïve and memory +cells have been shown to provide superior anti-tumor potency, presumably due to +their higher potential to replicate, migrate, and engraft, leading to a +long-term, durable response\cite{Xu2014, Fraietta2018, Gattinoni2011, + Gattinoni2012}. Likewise, CD4 T cells are similarly important to anti-tumor +potency due to their cytokine release properties and ability to resist +exhaustion\cite{Wang2018, Yang2017}. Therefore, methods to increase naïve/memory +and CD4 T cells in the final product are needed, a critical consideration being +ease of translation to industry and ability to interface with scalable systems +such as bioreactors. + +% TODO probably need to address some of the modeling stuff here as well + +This thesis describes a novel degradable microscaffold-based method derived from +porous microcarriers functionalized with anti-CD3 and anti-CD28 \glspl{mab} for +use in T cell expansion cultures. Microcarriers have historically been used +throughout the bioprocess industry for adherent cultures such as stem cells and +\gls{cho} cells, but not with suspension cells such as T +cells\cite{Heathman2015, Sart2011}. The microcarriers chosen to make the DMSs in +this study have a microporous structure that allows T cells to grow inside and +along the surface, providing ample cell-cell contact for enhanced autocrine and +paracrine signaling. Furthermore, the carriers are composed of gelatin, which is +a collagen derivative and therefore has adhesion domains that are also present +within the lymph nodes. Finally, the 3D surface of the carriers provides a +larger contact area for T cells to interact with the \glspl{mab} relative to +beads; this may better emulate the large contact surface area that occurs +between T cells and \glspl{dc}. These microcarriers are readily available in +over 30 countries and are used in an FDA fast-track-approved combination retinal +pigment epithelial cell product (Spheramine, Titan Pharmaceuticals) {\#}[Purcell +documentation]. This regulatory history will aid in clinical translation. We +show that compared to traditional microbeads, \gls{dms}-expanded T cells not +only provide superior expansion, but consistently provide a higher frequency of +naïve/memory and CD4 T cells (CCR7+CD62L+) across multiple donors. We also +demonstrate functional cytotoxicity using a CD19 \gls{car} and a superior +performance, even at a lower \gls{car} T cell dose, of \gls{dms}-expanded +\gls{car}-T cells in vivo in a mouse xenograft model of human B cell \gls{all}. +Our results indicate that \glspl{dms} provide a robust and scalable platform for +manufacturing therapeutic T cells with higher naïve/memory phenotype and more +balanced CD4+ T cell content. + \section*{overview} Insert overview here @@ -310,7 +397,7 @@ bla bla \chapter{References} \renewcommand{\section}[2]{} % noop the original bib section header -\bibliography{../proposal} +\bibliography{references} \bibliographystyle{naturemag}