Autophagy and apoptosis are catabolic pathways essential for organismal homeostasis. Autophagy is normally a cell-survival pathway involving the degradation and recycling of obsolete, damaged, or harmful macromolecular assemblies; however, excess autophagy has been implicated in type II cell death. Apoptosis is the canonical programmed cell death pathway. Autophagy and apoptosis have now been shown to be interconnected by several molecular nodes of crosstalk, enabling the coordinate regulation of degradation by these pathways. Normally, autophagy and apoptosis are both tumor suppressor pathways. Autophagy fulfils this role as it facilitates the degradation of oncogenic molecules, preventing development of cancers, while apoptosis prevents the survival of cancer cells. Consequently, defective or inadequate levels of either autophagy or apoptosis can lead to cancer. However, autophagy appears to have a dual role in cancer, as it has now been shown that autophagy also facilitates the survival of tumor cells in stress conditions such as hypoxic or low-nutrition environments. Here we review the multiple molecular mechanisms of coordination of autophagy and apoptosis and the role of the proteins involved in this crosstalk in cancer. A comprehensive understanding of the interconnectivity of autophagy and apoptosis is essential for the development of effective cancer therapeutics.

γ-herpesviruses (γHVs) are common human pathogens that encode homologs of the anti-apoptotic cellular Bcl-2 proteins, which are critical to viral reactivation and oncogenic transformation. The murine γHV68 provides a tractable in vivo model for understanding general features of these important human pathogens. Bcl-X_L, a cellular Bcl-2 homolog, and the murine γHV68 Bcl-2 homolog, M11, both bind to a BH3 domain within the key autophagy effector Beclin 1 with comparable affinities, resulting in the down-regulation of Beclin 1-mediated autophagy. Despite this similarity, differences in residues lining the binding site of M11 and Bcl-X_L dictate varying affinities for the different BH3 domain-containing proteins. Here we delineate Beclin 1 differential specificity determinants for binding to M11 or Bcl-X_L by quantifying autophagy levels in cells expressing different Beclin 1 mutants and either M11 or Bcl-X_L, and we show that a G120E/D121A Beclin 1 mutant selectively prevents down-regulation of Beclin 1-mediated autophagy by Bcl-X_L, but not by M11. We use isothermal titration calorimetry to identify a Beclin 1 BH3 domain-derived peptide that selectively binds to M11, but not to Bcl-X_L. The x-ray crystal structure of this peptide bound to M11 reveals the mechanism by which the M11 BH3 domain-binding groove accommodates this M11-specific peptide. This information was used to develop a cell-permeable peptide inhibitor that selectively inhibits M11-mediated, but not Bcl-X_L-mediated, down-regulation of autophagy.

Bottom-up approaches have emerged as a new philosophy in tissue engineering, enabling precise control over tissue morphogenesis at the cellular level. We previously prepared large bone-like tissues using cell-laden microbeads (microtissues) by following a modular approach to ensure cell viability. However, a long-term culture of such avascular macroscopic tissues (macrotissues) has not been evaluated. In the present study, microtissues were fabricated by cultivating human fibroblasts on Cytopore-2 microbeads in spinner flasks for 16 days. We then examined the long-term perfusion culture for macrotissues. Specifically, following assembly in a perfusion chamber for 15 days, cell death was found to be prominent at a depth of 500 µm from the surface of macrotissues towards the interior, suggesting that there was a new mass transfer limit leading to cell death instead of tissue maturation. Subsequently, we developed a strategy by incorporating microchannel structures in centimeter-sized tissue constructs to promote mass transport. By installing glass rods (1 mm diameter, 1 mm wall-to-wall spacing) in the perfusion chamber, stable microchannel architectures were introduced during the microtissue assembly process. Based on live/dead assay and scanning electron microscopy (SEM), these channelled macrotissues (length × diameter, 1.6 × 2.0 cm) demonstrated high cell viability and compact packing of microbeads. Comparative biochemical analysis further suggested a more homogeneous spatial distribution of cells and extracellular matrix (ECM) in the channelled macrotissues than in solid ones. Viable 3D large tissues can therefore be prepared by assembling cell-laden microbeads in conjunction with microchannel carving, meeting clinical needs in tissue repair. Copyright © 2012 John Wiley & Sons, Ltd.