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Summary Maintenance of body temperature is essential for survival of homeotherms. Brown adipose tissue (BAT) is a specialized fat tissue that is dedicated to thermoregulation1. Due to its remarkable capacity to dissipate stored energy and its demonstrated presence in adult humans2-5, BAT holds great promise for the treatment of obesity and metabolic syndrome1. Rodent data suggest the existence of two types of brown fat cells: the constitutive BAT (cBAT), which is of embryonic origin and anatomically located in the interscapular region of mice, and the recruitable BAT (rBAT) that resides within white adipose tissue (WAT)6 and skeletal muscle7, that has alternatively been called beige8, brite9, or inducible BAT10. Bone morphogenetic proteins (BMPs) regulate the formation and thermogenic activity of BAT10-12. We here provide evidence for a systemically active regulatory mechanism that serves to control whole body BAT-activity for thermoregulation and energy homeostasis. Genetic ablation of type 1A BMP-receptor (Bmpr1A) in brown adipogenic progenitor cells leads to a severe paucity of cBAT. This in turn increases sympathetic input to WAT, thereby promoting the formation of rBAT within white fat depots. This previously unknown compensatory mechanism, aimed at restoring total brown fat-mediated thermogenic capacity in the body, is sufficient to maintain normal temperature homeostasis and resistance to diet-induced obesity. These data suggest an important physiological cross-talk between the constitutive and recruitable brown fat cells. This sophisticated regulatory mechanism of body temperature may participate in the control of energy balance and metabolic disease.

Glioblastomas are lethal cancers defined by angiogenesis and pseudopalisading necrosis. Here, we demonstrate that these histological features are associated with distinct transcriptional programs, with vascular regions showing a proneural profile and hypoxic regions a mesenchymal pattern. As these regions harbor glioma stem cells (GSCs), we investigated the epigenetic regulation of these two niches. Proneural, perivascular GSCs activated EZH2, whereas mesenchymal GSCs in hypoxic regions expressed BMI1 protein, which promoted cellular survival under stress, due to downregulation of the E3 ligase, RNF144A. Using both genetic and pharmacologic inhibition, we found that proneural GSCs are preferentially sensitive to EZH2 disruption, whereas mesenchymal GSCs are preferentially sensitive to BMI1 inhibition. Given that glioblastomas contain both proneural and mesenchymal GSCs, combined EZH2 and BMI1 targeting proved more effective than either agent alone both in culture and in vivo, suggesting that strategies that simultaneously target multiple epigenetic regulators within glioblastomas may be necessary to overcome resistance to therapies caused by intratumoral heterogeneity.

The encapsulation of stem cells in a hydrogel substrate provides a promising future in biomedical applications. However, communications between hydrogels and stem cells is complicated; various factors such as porosity, different polymer types, stiffness, compatibility and degradation will lead to stem cell survival or death. Hydrogels mimic the three-dimensional extracellular matrix to provide a friendly environment for stem cells. On the other hand, stem cells can sense the surroundings to make the next progression, stretching out, proliferating or just to remain. As such, understanding the correlation between stem cells and hydrogels is crucial. In this Review, we first discuss the varying types of the hydrogels and stem cells, which are most commonly used in the biomedical fields and further investigate how hydrogels interact with stem cells from the perspective of their biomedical application, while providing insights into the design and development of hydrogels for drug delivery, tissue engineering and regenerative medicine purpose. In addition, we compare the results such as stiffness, degradation time and pore size as well as peptide types of hydrogels from respected journals. We also discussed most recently magnificent materials and their effects to regulate stem cell fate.

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Phospholipase D (PLD) is a key facilitator of multiple types of membrane vesicle trafficking events. Two PLD isoforms, PLD1 and PLD2, exist in mammals. Initial studies based on overexpression studies suggested that in resting cells, human PLD1 localized primarily to the Golgi and perinuclear vesicles in multiple cell types. In contrast, overexpressed mouse PLD2 was observed to localize primarily to the plasma membrane, although internalization on membrane vesicles was observed subsequent to serum stimulation. A recent report has suggested that the assignment of PLD2 to the plasma membrane is in error, because the endogenous isoform in rat secretory cells was imaged and found to be present primarily in the Golgi apparatus. We have reexamined this issue by using a monoclonal antibody specific for mouse PLD2, and find, as reported initially using overexpression studies, that endogenous mouse PLD2 is detected most readily at the plasma membrane in multiple cell types. In addition, we report that mouse, rat, and human PLD2 when overexpressed all similarly localize to the plasma membrane in cell lines from all three species. Finally, studies conducted using overexpression of wild-type active or dominant-negative isoforms of PLD2 and RNA interference-mediated targeting of PLD2 suggest that PLD2 functions at the plasma membrane to facilitate endocytosis of the angiotensin II type 1 receptor. INTRODUCTIONPhospholipase D (PLD), which hydrolyzes phosphatidylcholine to generate choline and the bioactive lipid phosphatidic acid, has been implicated in signal transduction, membrane trafficking, transformation, and cytoskeletal reorganization (reviewed in Frohman et al., 1999;Jones et al., 1999;Liscovitch et al., 2000). There are two mammalian PLDs, PLD1 (Hammond et al., 1995) and PLD2 (Colley et al., 1997c), which are expressed in a wide but selective variety of tissues and cells (Colley et al., 1997a; reviewed in Gibbs and Meier, 2000). Although frequently expressed in the same cell types, PLD1 and PLD2 have generally been proposed to mediate isoform-specific functions, based on their selective abilities, when overexpressed as wild-type or catalytically inactive alleles, to facilitate or hinder different steps in cytoskeletal reorganization (Colley et al., 1997b;O'Luanaigh et al., 2002) and regulated exocytosis from neuroendocrine cells (Vitale et al., 2001) and mast cells (Choi et al., 2002).In addition, numerous reports based on overexpression have described localization of the isoforms to separate regions of the cell-PLD1 to perinuclear vesicles (Colley et al., 1997b;Toda et al., 1999;Lucocq et al., 2001)-and PLD2 to the plasma membrane (Colley et al., 1997b;Park et al., 2000;O'Luanaigh et al., 2002). Localization of the isoforms is clearly complex though. In some cells, PLD1 preferentially localizes to the plasma membrane (Vitale et al., 2001), and more generally, it has been shown to cycle between perinuclear regions and the plasma membrane (Brown et al., 1998;Emoto et al., 2000;Du et al., 2003). Similarly, internaliza...

Glioblastoma (GBM) contains a self-renewing, tumorigenic cancer stem cell (CSC) population which contributes to tumor propagation and therapeutic resistance. While the tumor microenvironment is essential to CSC self-renewal, the mechanisms by which CSCs sense and respond to microenvironmental conditions are poorly understood. Scavenger receptors are a broad class of membrane receptors that are well characterized on immune cells and instrumental in sensing apoptotic cellular debris and modified lipids. Here we provide evidence that CSCs selectively utilize the scavenger receptor CD36 to promote their maintenance using patient-derived CSCs and in vivo xenograft models. We detected CD36 expression in GBM cells in addition to previously described cell types including endothelial cells, macrophages and microglia. CD36 was enriched in CSCs and was able to functionally distinguish self-renewing cells. CD36 was co-expressed with integrin alpha 6 and CD133, previously described CSC markers, and CD36 reduction resulted in concomitant loss of integrin alpha 6 expression, self-renewal and tumor initiation capacity. We confirmed that oxidized phospholipids, ligands of CD36, were present in GBM and found that the proliferation of CSCs, but not non-CSCs, increased with exposure to oxidized low-density lipoprotein. CD36 was an informative biomarker of malignancy and negatively correlated to patient prognosis. These results provide a paradigm for CSCs to thrive by the selective enhanced expression of scavenger receptors, providing survival and metabolic advantages.

The signaling enzyme phospholipase D1 (PLD1) facilitates membrane vesicle trafficking. Here, we explore how PLD1 subcellular localization is regulated via Phox homology (PX) and pleckstrin homology (PH) domains and a PI4,5P2-binding site critical for its activation. PLD1 localized to perinuclear endosomes and Golgi in COS-7 cells, but on cellular stimulation, translocated to the plasma membrane in an activity-facilitated manner and then returned to the endosomes. The PI4,5P2-interacting site sufficed to mediate outward translocation and association with the plasma membrane. However, in the absence of PX and PH domains, PLD1 was unable to return efficiently to the endosomes. The PX and PH domains appear to facilitate internalization at different steps. The PH domain drives PLD1 entry into lipid rafts, which we show to be a step critical for internalization. In contrast, the PX domain appears to mediate binding to PI5P, a lipid newly recognized to accumulate in endocytosing vesicles. Finally, we show that the PH domain–dependent translocation step, but not the PX domain, is required for PLD1 to function in regulated exocytosis in PC12 cells. We propose that PLD1 localization and function involves regulated and continual cycling through a succession of subcellular sites, mediated by successive combinations of membrane association interactions.