Shared Transformation

A conversation with Lisa Iwamoto of IwamotoScott Architectur

Jellyfish House

This paper intoduces Jellyfish House, designed for the exhibition OPEN HOUSE: Architecture and Technology for Intelligent Living curated by the Vitra Design Museum and Art Center College of Design. Conceptually, the house draws from ‘calm technology,’ a branch of research associated with ubiquitous computing. Calm technology suggests that the digital realm will recede to the background of our spaces and lived experience

Autophagy-mediated apoptosis eliminates aneuploid cells in a mouse model of chromosome mosaicism

Funder: Open Philanthropy grants to M.Z.G.Abstract: The high incidence of aneuploidy in the embryo is considered the principal cause for low human fecundity. However, the prevalence of aneuploidy dramatically declines as pregnancy progresses, with the steepest drop occurring as the embryo completes implantation. Despite the fact that the plasticity of the embryo in dealing with aneuploidy is fundamental to normal development, the mechanisms responsible for eliminating aneuploid cells are unclear. Here, using a mouse model of chromosome mosaicism, we show that aneuploid cells are preferentially eliminated from the embryonic lineage in a p53-dependent process involving both autophagy and apoptosis before, during and after implantation. Moreover, we show that diploid cells in mosaic embryos undertake compensatory proliferation during the implantation stages to confer embryonic viability. Together, our results indicate a close link between aneuploidy, autophagy, and apoptosis to refine the embryonic cell population and ensure only chromosomally fit cells proceed through development of the fetus

Autophagy-mediated apoptosis eliminates aneuploid cells in a mouse model of chromosome mosaicism

Funder: Open Philanthropy grants to M.Z.G.Abstract: The high incidence of aneuploidy in the embryo is considered the principal cause for low human fecundity. However, the prevalence of aneuploidy dramatically declines as pregnancy progresses, with the steepest drop occurring as the embryo completes implantation. Despite the fact that the plasticity of the embryo in dealing with aneuploidy is fundamental to normal development, the mechanisms responsible for eliminating aneuploid cells are unclear. Here, using a mouse model of chromosome mosaicism, we show that aneuploid cells are preferentially eliminated from the embryonic lineage in a p53-dependent process involving both autophagy and apoptosis before, during and after implantation. Moreover, we show that diploid cells in mosaic embryos undertake compensatory proliferation during the implantation stages to confer embryonic viability. Together, our results indicate a close link between aneuploidy, autophagy, and apoptosis to refine the embryonic cell population and ensure only chromosomally fit cells proceed through development of the fetus

IL-17A Expression Is Localised to Both Mononuclear and Polymorphonuclear Synovial Cell Infiltrates

This study examines the expression of IL-17A-secreting cells within the inflamed synovium and the relationship to in vivo joint hypoxia measurements.IL-17A expression was quantified in synovial tissue (ST), serum and synovial fluid (SF) by immunohistochemistry and MSD-plex assays. IL-6 SF and serum levels were measured by MSD-plex assays. Dual immunofluorescence for IL-17A was quantified in ST CD15+ cells (neutrophils), Tryptase+ (mast cells) and CD4+ (T cells). Synovial tissue oxygen (tpO(2)) levels were measured under direct visualisation at arthroscopy. Synovial infiltration was assessed using immunohistochemistry for cell specific markers. Peripheral blood mononuclear and polymorphonuclear cells were isolated and exposed to normoxic or 3% hypoxic conditions. IL-17A and IL-6 were quantified as above in culture supernatants.IL-17A expression was localised to mononuclear and polymorphonuclear (PMN) cells in inflamed ST. Dual immunoflourescent staining co-localised IL-17A expression with CD15+ neutrophils Tryptase+ mast cells and CD4+T cells. % IL-17A positivity was highest on CD15+ neutrophils, followed by mast cells and then CD4+T-cells. The number of IL-17A-secreting PMN cells significantly correlated with sublining CD68 expression (r = 0.618, p<0.01). IL-17A SF levels correlated with IL-6 SF levels (r = 0.675, p<0.01). Patients categorized according to tp0(2)< or >20 mmHg, showed those with low tp0(2)<20 mmHg had significantly higher IL-17A+ mononuclear cells with no difference observed for PMNs. Exposure of mononuclear and polymorphonuclear cells to 3% hypoxia, significantly induced IL-6 in mononuclear cells, but had no effect on IL-17A expression in mononuclear and polymorphonuclear cells.This study demonstrates IL-17A expression is localised to several immune cell subtypes within the inflamed synovial tissue, further supporting the concept that IL-17A is a key mediator in inflammatory arthritis. The association of hypoxia with Il-17A expression appears to be indirect, probably through hypoxia-induced pro-inflammatory pathways and leukocyte influx within the joint microenvironment

Digital fabrications : architectural and material techniques

144 p. : col. ill. ; 24 cm

Digital fabrications : architectural and material techniques

144hlm.;bib.;ill

Cell Fate Decisions in the Early Mammalian Embryo

In mammals, successful pre-implantation development leads to the formation of a tri-lineage structure known as the blastocyst, consisting of the epiblast, primitive endoderm and trophectoderm, which will give rise to the new organism, the yolk sac and placenta respectively. These three lineages must be established from the totipotent zygote via two successive cell fate decisions in the appropriate sequence, position and proportion, to generate a blastocyst capable of implantation and further development. In mammals this process has long been thought to be regulative, with cell-cell interactions flexibly determining the eventual fate of cell. This is in contrast to commonly studied non-mammalian embryos in which pre-patterning of the embryo, driven by spatially localised factors, is a common feature. However, early blastomeres of mouse embryos have been reported to have distinct developmental fates, potential and heterogeneous abundance of certain transcripts, prior to the first cell fate decision. Nevertheless, the extent of the earliest intra-embryo differences remains unclear and controversial. Utilizing single-cell proteomics by mass-spectrometry I show that 2-cell mouse and human embryos contain an alpha and a beta blastomere as defined by differential abundance of hundreds of proteins. Such asymmetrically distributed proteins include Gps1 and Nedd8, depletion or overexpression of which in one blastomere of the 2-cell embryo impacts lineage segregation. Fascinatingly, halved mouse zygotes already display protein asymmetries, which resembles alpha and beta blastomeres, suggesting differential proteome localisation already within zygotes. I also find that beta blastomeres may have a greater developmental potential, and give rise to a blastocyst with a higher proportion of epiblast cells than alpha blastomeres. Human 2-cell blastomeres also partition into two clusters sharing strong concordance with clusters found in mouse, in terms of differentially abundant proteins and functional enrichment. This provides the first demonstration of intra-zygotic and inter-blastomere proteomic asymmetry in mammals that has a role in lineage segregation. In humans, this early period of development is prone to failure, with a third of human pregnancies estimated as being lost prior to implantation. A high incidence of aneuploidy is thought to be a major driver of pregnancy failure and understanding the behaviour of aneuploid cells is of great interest. As the blastocyst forms and implants, aneuploid cells may be eliminated through cell competition with diploid cells or show differences in their lineage segregation, impacting the composition and proportion of each lineage in the blastocyst, The mouse embryo does not have similar high intrinsic rates of aneuploidy, but reversine, a spindle assembly checkpoint inhibitor, can be used to recapitulate the human aneuploid embryo to some extent, and to observe the elimination of aneuploid cells. Here, I return to the human, utilising human embryonic stem cells and new integrated stem cell based embryo models, to characterise conserved aneuploid cell depletion in mouse and human stem cell co-cultures of diploid and aneuploid cells. Furthermore, I use stem cell lines harbouring specific aneuploidies to determine if specific aneuploidies confer differential ‘fitness’ and elimination rates. Overall my PhD has examined two questions regarding early mammalian development: 1) when do the cells of the embryo first become different to each other and how does this interplay with lineage segregation and 2) can a model of the mosaic aneuploid human embryo be generated to better understand the fate of aneuploid cells within the three lineages of the blastocyst