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Organogenesis is the phase of embryonic development that starts at the end of gastrulation and continues until birth. During organogenesis, the three germ layers formed from gastrulation (the ectoderm, endoderm, and mesoderm) form the internal organs of the organism. The cells of each of the three germ layers undergo differentiation, a process where less-specialized cells become more-specialized through the expression of a specific set of genes. Cell differentiation is driven by cell signaling cascades. Differentiation is influenced by extracellular signals such as growth factors that are exchanged to adjacent cells which is called juxtracrine signaling or to neighboring cells over short distances which is called paracrine signaling. Intracellular signals consist of a cell signaling itself (autocrine signaling), also play a role in organ formation. These signaling pathways allow for cell rearrangement and ensure that organs form at specific sites within the organism. The organogenesis process can be studied using embryos and organoids. The endoderm is the inner most germ layer of the embryo which gives rise to gastrointestinal and respiratory organs by forming epithelial linings and organs such as the liver, lungs, and pancreas. The mesoderm or middle germ layer of the embryo will form the blood, heart, kidney, muscles, and connective tissues. The ectoderm or outermost germ layer of the developing embryo forms epidermis, the brain, and the nervous system. While each germ layer forms specific organs, in the 1820s, embryologist Heinz Christian Pander discovered that the germ layers cannot form their respective organs without the cellular interactions from other tissues. In humans, internal organs begin to develop within 3–8 weeks after fertilization. The germ layers form organs by three processes: folds, splits, and condensation. Folds form in the germinal sheet of cells and usually form an enclosed tube which you can see in the development of vertebrates neural tube. Splits or pockets may form in the germinal sheet of cells forming vesicles or elongations.

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Organoids are powerful models for studying tissue development, physiology, and disease. However, current culture systems disrupt the inductive tissue-tissue interactions needed for the complex morphogenetic processes of native organogenesis. Here, we show that mouse embryonic stem cells (mESCs) can be coaxed to robustly undergo fundamental steps of early heart organogenesis with an in-vivo-like spatiotemporal fidelity. These axially patterned embryonic organoids (gastruloids) mimic embryonic development and support the generation of cardiovascular progenitors, including first and second heart fields. The cardiac progenitors self-organize into an anterior domain reminiscent of a cardiac crescent before forming a beating cardiac tissue near a putative primitive gut-like tube, from which it is separated by an endocardial-like layer. These findings unveil the surprising morphogenetic potential of mESCs to execute key aspects of organogenesis through the coordinated development of multiple tissues. This platform could be an excellent tool for studying heart development in unprecedented detail and throughput.

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Overexpression of transcription factor Foxa1 and target genes remediate therapeutic protein production bottlenecks in Chinese hamster ovary cells

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Despite extensive research conducted to increase protein production from Chinese hamster ovary (CHO) cells, cellular bottlenecks often remain, hindering high yields. In this study, a transcriptomic analysis led to the identification of 32 genes that are consistently upregulated in high producer clones and thus might mediate high productivity. Candidate genes were associated with functions such as signaling, protein folding, cytoskeleton organization, and cell survival. We focused on two engineering targets, Erp27, which binds unfolded proteins and the Erp57 disulfide isomerase in the endoplasmic reticulum, and Foxa1, a pioneering transcription factor involved in organ development. Erp27 moderate overexpression increased production of an easy-to-express antibody, whereas Erp27 and Erp57 co-overexpression increased cell density, viability, and the yield of difficult-to-express proteins. Foxa1 overexpression increased cell density, cell viability, and easy- and difficult-to-express protein yields, whereas it decreased reactive oxygen species late in fed-batch cultures. Foxa1 overexpression upregulated two other candidate genes that increased the production of difficult- and/or easy-to-express proteins, namely Ca3, involved in protecting cells from oxidative stress, and Tagap, involved in signaling and cytoskeleton remodeling. Overall, several genes allowing to overcome CHO cell bottlenecks were identified, including Foxa1, which mediated multiple favorable metabolic changes that improve therapeutic protein yields.

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