MafB restricts M-CSF-dependent myeloid commitment divisions of hematopoietic stem cells.

While hematopoietic stem cell (HSC) self-renewal is well studied, it remains unknown whether distinct control mechanisms enable HSC divisions that generate progeny cells with specific lineage bias. Here, we report that the monocytic transcription factor MafB specifically restricts the ability of M-CSF to instruct myeloid commitment divisions in HSCs. MafB deficiency specifically enhanced sensitivity to M-CSF and caused activation of the myeloid master-regulator PU.1 in HSCs in vivo. Single-cell analysis revealed that reduced MafB levels enabled M-CSF to instruct divisions producing asymmetric daughter pairs with one PU.1(+) cell. As a consequence, MafB(-/-) HSCs showed a PU.1 and M-CSF receptor-dependent competitive repopulation advantage specifically in the myelomonocytic, but not T lymphoid or erythroid, compartment. Lineage-biased repopulation advantage was progressive, maintained long term, and serially transplantable. Together, this indicates that an integrated transcription factor/cytokine circuit can control the rate of specific HSC commitment divisions without compromising other lineages or self-renewal.

JAK2V617F homozygosity drives a phenotypic switch in myeloproliferative neoplasms, but is insufficient to sustain disease.

Genomic regions of acquired uniparental disomy (UPD) are common in malignancy and frequently harbor mutated oncogenes. Homozygosity for such gain-of-function mutations is thought to modulate tumor phenotype, but direct evidence has been elusive. Polycythemia vera (PV) and essential thrombocythemia (ET), 2 subtypes of myeloproliferative neoplasms, are associated with an identical acquired JAK2V617F mutation but the mechanisms responsible for distinct clinical phenotypes remain unclear. We provide direct genetic evidence and demonstrate that homozygosity for human JAK2V617F in knock-in mice results in a striking phenotypic switch from an ET-like to PV-like phenotype. The resultant erythrocytosis is driven by increased numbers of early erythroid progenitors and enhanced erythroblast proliferation, whereas reduced platelet numbers are associated with impaired platelet survival. JAK2V617F-homozygous mice developed a severe hematopoietic stem cell defect, suggesting that additional lesions are needed to sustain clonal expansion. Together, our results indicate that UPD for 9p plays a causal role in the PV phenotype in patients as a consequence of JAK2V617F homozygosity. The generation of a JAK2V617F allelic series of mice with a dose-dependent effect on hematopoiesis provides a powerful model for studying the consequences of mutant JAK2 homozygosity.

Development and Characterization of Polymeric-based Biomaterial from Agro-food Waste: A Sustainable and Eco-friendly Approach Towards Plastic Pollution

INTRODUCTION: Commercial plastics are potentially hazardous and can be carcinogenic due to the incorporation of chemical additives along with other additional components utilized as brominated flame retardants and phthalate plasticizers during production that excessively produce large numbers of gases, litter, and toxic components resulting in environmental pollution. METHOD: Biodegradable plastic derived from natural renewable resources is the novel, alternative, and innovative approach considered to be potentially safe as a substitute for traditional synthetic plastic as they decompose easily without causing any harm to the ecosystem and natural habitat. The utilization of undervalued compounds, such as by-products of fruits and vegetables in the production of biodegradable packaging films, is currently a matter of interest because of their accessibility, affordability, ample supply, nontoxicity, physiochemical and nutritional properties. Industrial food waste was processed under controlled conditions with appropriate plasticizers to extract polymeric materials. Biodegradability, solubility, and air test analysis were performed to examine the physical properties of polymers prior to the characterization of the biofilm by Fourier-transformed infrared spectroscopy (FTIR) for the determination of polymeric characteristics. RESULT: The loss of mass examined in each bioplastic film was in the range of 0.01g to 0.20g. The dimension of each bioplastic was recorded in the range of 4.6 mm to 28.7 mm. The existence of -OH, C=C, C=O stretching, and other crucial functional groups that aid in the creation of a solid polymeric material are confirmed by FTIR analysis. This study provides an alternative approach for sustainable and commercially value-added production of polymeric-based biomaterials from agro-industrial waste as they are rich in starch, cellulose, and pectin for the development of bio-plastics. CONCLUSION: The rationale of this project is to achieve a straightforward, economical, and durable method for the production of bio-plastics through effective utilization of industrial and commercial fruit waste, ultimately aiding in revenue generation.

Circular and linear: a tale of aptamer selection for the activation of SIRT1 to induce death in cancer cells.

It is a challenge to select the right target to treat conditions without affecting non-diseased cells. Cancer belongs to the top 10 causes of death in the world and it remains difficult to treat. Amongst cancer emerging targets, silent information regulator 1 (SIRT1) - a histone deacetylase - has shown many roles in cancer, ageing and metabolism. Here we report novel SIRT1 ligands that bind and modulate the activity of SIRT1 within cells and enhance its enzymatic activity. We developed a modified aptamer capable of binding to and forming a complex with SIRT1. Our ligands are aptamers, they can be made of DNA or RNA oligonucleotides, their binding domain can recognise a target with very high affinity and specificity. We used the systematic evolution of ligands by exponential enrichment (SELEX) technique to develop circular and linear aptamers selectively binding to SIRT1. Cellular consequences of the interaction were monitored by fluorescence microscopy, cell viability assay, stability and enzymatic assays. Our results indicate that from our pool of aptamers, circular AC3 penetrates cancerous cells and is recruited to modulate the SIRT1 activity. This modulation of SIRT1 resulted in anticancer activity on different cancer cell lines. Furthermore, this modified aptamer showed no toxicity on one non-cancerous cell line and was stable in human plasma. We have demonstrated that aptamers are efficient tools for localisation of internal cell targets, and in this particular case, anticancer activity through modulation of SIRT1.

Development of Polymeric Nanocomposite (Xyloglucan-co-Methacrylic Acid/Hydroxyapatite/SiO2) Scaffold for Bone Tissue Engineering Applications-In-Vitro Antibacterial, Cytotoxicity and Cell Culture Evaluation.

Advancement and innovation in bone regeneration, specifically polymeric composite scaffolds, are of high significance for the treatment of bone defects. Xyloglucan (XG) is a polysaccharide biopolymer having a wide variety of regenerative tissue therapeutic applications due to its biocompatibility, in-vitro degradation and cytocompatibility. Current research is focused on the fabrication of polymeric bioactive scaffolds by freeze drying method for nanocomposite materials. The nanocomposite materials have been synthesized from free radical polymerization using n-SiO2 and n-HAp XG and Methacrylic acid (MAAc). Functional group analysis, crystallinity and surface morphology were investigated by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction analysis (XRD) and scanning electron microscopy (SEM) techniques, respectively. These bioactive polymeric scaffolds presented interconnected and well-organized porous morphology, controlled precisely by substantial ratios of n-SiO2. The swelling analysis was also performed in different media at varying temperatures (27, 37 and 47 °C) and the mechanical behavior of the dried scaffolds is also investigated. Antibacterial activities of these scaffolds were conducted against pathogenic gram-positive and gram-negative bacteria. Besides, the biological behavior of these scaffolds was evaluated by the Neutral Red dye assay against the MC3T3-E1 cell line. The scaffolds showed interesting properties for bone tissue engineering, including porosity with substantial mechanical strength, biodegradability, biocompatibility and cytocompatibility behavior. The reported polymeric bioactive scaffolds can be aspirant biomaterials for bone tissue engineering to regenerate defecated bone.

Development of macrophages with altered actin organization in the absence of MafB.

In the hematopoietic system the bZip transcription factor MafB is selectively expressed at high levels in monocytes and macrophages and promotes macrophage differentiation in myeloid progenitors, whereas a dominant-negative allele can inhibit this process. To analyze the requirement of MafB for macrophage development, we generated MafB-deficient mice and, due to their neonatal lethal phenotype, analyzed macrophage differentiation in vitro, in the embryo, and in reconstituted mice. Surprisingly we observed in vitro differentiation of macrophages from E14.5 fetal liver (FL) cells and E18.5 splenocytes. Furthermore we found normal numbers of F4/80(+)/Mac-1(+) macrophages and monocytes in fetal liver, spleen, and blood as well as in bone marrow, spleen, and peritoneum of adult MafB(-/-) FL reconstituted mice. MafB(-/-) macrophages showed intact basic macrophage functions such as phagocytosis of latex beads or Listeria monocytogenes and nitric oxide production in response to lipopolysaccharide. By contrast, MafB(-/-) macrophages expressed increased levels of multiple genes involved in actin organization. Consistent with this, phalloidin staining revealed an altered morphology involving increased numbers of branched protrusions of MafB(-/-) macrophages in response to macrophage colony-stimulating factor. Together these data point to an unexpected redundancy of MafB function in macrophage differentiation and a previously unknown role in actin-dependent macrophage morphology.

Dibenzoyl-methane Derivatives as a Potential and Exciting New Therapy for the Treatment of Childhood Bone Cancer.

Childhood bone cancer though rare, has very limited treatment choices, with poor survival rates and often involving amputation. We developed a novel molecule, 2', 4'-dihydroxy-dithion-dibenzoyl-methane and tested it on hepatic, colon, lung and osteoblast cancer cell lines. Thionylation of 2', 4'- dihydroxydibenzoylmethane led to selective targeting of bone cancer cells, stopping their growth and leading to their death without affecting non-cancerous cells within the bone marrow or other non-malignant cells.

Cooperativity of imprinted genes inactivated by acquired chromosome 20q deletions.

Large regions of recurrent genomic loss are common in cancers; however, with a few well-characterized exceptions, how they contribute to tumor pathogenesis remains largely obscure. Here we identified primate-restricted imprinting of a gene cluster on chromosome 20 in the region commonly deleted in chronic myeloid malignancies. We showed that a single heterozygous 20q deletion consistently resulted in the complete loss of expression of the imprinted genes L3MBTL1 and SGK2, indicative of a pathogenetic role for loss of the active paternally inherited locus. Concomitant loss of both L3MBTL1 and SGK2 dysregulated erythropoiesis and megakaryopoiesis, 2 lineages commonly affected in chronic myeloid malignancies, with distinct consequences in each lineage. We demonstrated that L3MBTL1 and SGK2 collaborated in the transcriptional regulation of MYC by influencing different aspects of chromatin structure. L3MBTL1 is known to regulate nucleosomal compaction, and we here showed that SGK2 inactivated BRG1, a key ATP-dependent helicase within the SWI/SNF complex that regulates nucleosomal positioning. These results demonstrate a link between an imprinted gene cluster and malignancy, reveal a new pathogenetic mechanism associated with acquired regions of genomic loss, and underline the complex molecular and cellular consequences of "simple" cancer-associated chromosome deletions.

MafB/c-Maf deficiency enables self-renewal of differentiated functional macrophages.

In metazoan organisms, terminal differentiation is generally tightly linked to cell cycle exit, whereas the undifferentiated state of pluripotent stem cells is associated with unlimited self-renewal. Here, we report that combined deficiency for the transcription factors MafB and c-Maf enables extended expansion of mature monocytes and macrophages in culture without loss of differentiated phenotype and function. Upon transplantation, the expanded cells are nontumorigenic and contribute to functional macrophage populations in vivo. Small hairpin RNA inactivation shows that continuous proliferation of MafB/c-Maf deficient macrophages requires concomitant up-regulation of two pluripotent stem cell-inducing factors, KLF4 and c-Myc. Our results indicate that MafB/c-MafB deficiency renders self-renewal compatible with terminal differentiation. It thus appears possible to amplify functional differentiated cells without malignant transformation or stem cell intermediates.