The metabolically-modulated stem cell niche: a dynamic scenario regulating cancer cell phenotype and resistance to therapy

This Perspective addresses the interactions of cancer stem cells (CSC) with environment which result in the modulation of CSC metabolism, and thereby of CSC phenotype and resistance to therapy. We considered first as a model disease chronic myeloid leukemia (CML), which is triggered by a well-identified oncogenetic protein (BCR/Abl) and brilliantly treated with tyrosine kinase inhibitors (TKi). However, TKi are extremely effective in inducing remission of disease, but unable, in most cases, to prevent relapse. We demonstrated that the interference with cell metabolism (oxygen/glucose shortage) enriches cells exhibiting the leukemia stem cell (LSC) phenotype and, at the same time, suppresses BCR/Abl protein expression. These LSC are therefore refractory to the TKi Imatinib-mesylate, pointing to cell metabolism as an important factor controlling the onset of TKi-resistant minimal residual disease (MRD) of CML and the related relapse. Studies of solid neoplasias brought another player into the control of MRD, low tissue pH, which often parallels cancer growth and progression. Thus, a 3-party scenario emerged for the regulation of CSC/LSC maintenance, MRD induction and disease relapse: the “hypoxic” versus the “ischemic” vs. the “acidic” environment. As these environments are unlikely constrained within rigid borders, we named this model the “metabolically-modulated stem cell niche.”     

Src Homology 2 Domain Containing Protein 5 (SH2D5) Binds the Breakpoint Cluster Region Protein,BCR, and Regulates Levels of Rac1-GTP

SH2D5 is a mammalian-specific, uncharacterized adaptor-like protein that contains an N-terminal phosphotyrosine-binding domain and a C-terminal Src homology 2 (SH2) domain. We show that SH2D5 is highly enriched in adult mouse brain, particularly in Purkinjie cells in the cerebellum and the cornu ammonis of the hippocampus. Despite harboring two potential phosphotyrosine (Tyr(P)) recognition domains, SH2D5 binds minimally to Tyr(P) ligands, consistent with the absence of a conserved Tyr(P)-binding arginine residue in the SH2 domain. Immunoprecipitation coupled to mass spectrometry (IP-MS) from cultured cells revealed a prominent association of SH2D5 with breakpoint cluster region protein, a RacGAP that is also highly expressed in brain. This interaction occurred between the phosphotyrosine-binding domain of SH2D5 and an NxxF motif located within the N-terminal region of the breakpoint cluster region. siRNA-mediated depletion of SH2D5 in a neuroblastoma cell line, B35, induced a cell rounding phenotype correlated with low levels of activated Rac1-GTP, suggesting that SH2D5 affects Rac1-GTP levels. Taken together, our data provide the first characterization of the SH2D5 signaling protein.     

PHYTOEXTRACTION OF METAL POLLUTED SOILS AROUND A Pb-Zn MINE BY CROP PLANTS

In order to assess their practical capability for the absorption and accumulation of Pb, Zn, and Cu, five common crop plants, i.e. maize (Zea mays), sunflower (Helianthus annuus), canola (Brassica napus), barley (Hordeum vulgare) and White lupine (Lupinus albus) were tested in pot experiments using six soil samples taken from mine tailings, pasture and arable soils around an old Pb-Zn mine in Spain. Metal concentration ranges of the soils were 76.2–785 mg kg^?1, 127–1652 mg kg^?1, and 12.4–82.6 mg kg^?1 for Zn, Pb, and Cu, respectively. With the exception of the highest polluted sample, soil total metal concentration did not influence significantly biomass yields of each crop for the different growth substrates. The order found for the total metal accumulation rate (TMAR) in the crops was Zn>>Pb > Cu, with maize reaching the highest metal concentrations. Pb root concentrations were markedly higher than those of shoots for all the crops, while Zn and Cu were translocated to shoots more efficiently. Concentrations of metals extracted by EDTA and BCR sequential extraction were well correlated, in general, with both root metal content and TMAR. CaCl₂-extracted Zn was well correlated with root concentrations, TMAR and, in some cases, with shoot contents. Our study showed that the test crops were not feasible to remediate the heavily or moderately contaminated soils studied here in order to achieve the total metal soil concentrations required by the current European laws.     

湘西河流表层沉积物重金属污染特征及其潜在生态毒性风险

花垣河和峒河是湘西地区受到锰矿和铅锌矿生产影响严重的两条河流。通过表层沉积物采样分析了Cd、Pb、Cu、Ni、Cr、Zn和Mn的总量,根据BCR连续提取程序分析沉积物样品中重金属的地球化学赋存形态,采用内梅罗指数法和地积累指数法评价了沉积物重金属污染特征,根据重金属的富集程度探讨了重金属污染来源,采用淡水生态系统沉积物质量基准(SQGs,TEL/PEL)和毒性单位评价了花垣河和峒河沉积物中重金属元素的生态毒性风险。结果表明,花垣河和峒河绝大多数位点的表层沉积物中Cd、Pb、Cu、Ni、Cr、Zn和Mn的总量高于参照点,形成严重的复合污染,花垣河沉积物中重金属的污染水平明显高于峒河,但沿程变化规律不明显,而峒河沉积物中重金属的沿程变化较有规律,即上游含量低,中下游含量较高。两条河流表层沉积物中富集程度居前列的均为Cd、Pb、Zn和Mn。花垣河和峒河沉积物重金属污染主要来源于矿业生产所产生废渣和废水的点排放。在花垣河和峒河的大多数位点,Cd、Pb和Mn的形态具有共同特征,其生物可利用态均较大程度地超过生物不可利用态,而且Mn和Cd的生物可直接利用态所占比例远高于其它重金属,而Cu和Cr的生物可直接利用态所占比例很低。花垣河沉积物中Cd、Pb和Zn在所有位点极大地超过PEL,在峒河中下游,Cd、Pb、Ni和Zn超过PEL,具有较大的潜在生物毒性。除上游S1位点外,花垣河的其余各位点都具有明显的急性毒性,峒河中下游各位点具有明显的急性毒性,这些河段需要重点治理。     

Integration of a Phosphatase Cascade with the Mitogen-activated Protein Kinase Pathway Provides for a Novel Signal Processing Function

We mathematically modeled the receptor-dependent mitogen-activated protein kinase (MAPK) signaling by incorporating the regulation through cellular phosphatases. Activation induced the alignment of a phosphatase cascade in parallel with the MAPK pathway. A novel regulatory motif was, thus, generated, providing for the combinatorial control of each MAPK intermediate. This ensured a non-linear mode of signal transmission with the output being shaped by the balance between the strength of input signal and the activity gradient along the phosphatase axis. Shifts in this balance yielded modulations in topology of the motif, thereby expanding the repertoire of output responses. Thus, we identify an added dimension to signal processing wherein the output response to an external stimulus is additionally filtered through indicators that define the phenotypic status of the cell.     

Soluble HLA/peptide monomers cross-linked with co-stimulatory antibodies onto a streptavidin core molecule efficiently stimulate antigen-specific T cell responses

Soluble MHC–peptide complexes, commonly referred to as tetramers, have been shown to induce strong cross-linking of TCR and CD8, resulting in a vigorous activation followed by a rapid non-apoptotic CD8⁺ T cell death. This has limited tetramer use for antigen-specific T cells isolation and cloning, as sorted tetramer positive cells were shown to possess compromised functional integrity. Here we show that the cross-linking of a secondary co-stimulatory signal into oligomeric MHC:peptide complexes prevents such cell death, and in contrast strongly stimulates antigen-specific T cell responses. Such soluble antigen-presenting complexes (sAPCs) containing MHC:peptide complexes linked to either anti-CD27 or anti-CD28 antibodies were capable of priming and expanding HLA-A*0201 restricted CMV specific T cells and also of generating functional HLA-A*0301 restricted BCR/ABL-specific T cell responses. These sAPCs constitute an encouraging alternative method for generating antigen-specific T cells that could be applied to a variety of antigens. A. I. Dodi and P. J. Travers contributed equally to this work. This paper is an original contribution from the meeting which took place on 28 and 29 May 2008 in Nottingham, UK, celebrating the contribution of Prof. I. A. “Tony” Dodi (^†29.1.2008) to the EU project “Network for the identification and validation of antigens and biomarkers in cancer and their application in clinical tumour immunology (ENACT)”.     

Centromeric interval of chromosome 4 derived from C57BL/6 mice accelerates type 1 diabetes in NOD.CD72 congenic mice

The nonobese diabetic (NOD) mouse is a useful model of autoimmune type 1 diabetes exhibiting many similarities to human type 1 diabetes patients including the presence of auto-reactive T cells and pancreas-specific autoantiboies. Multiple Idd loci control the development of diabetes in NOD mice. CD72, a B cell membrane-bound glycoprotein carrying a C-type lectin-like domain, is an inhibitory co-receptor of the B cell antigen receptor (BCR) that negatively regulates BCR signaling. Among four known haplotypes of mouse CD72, NOD mice carry the CD72^c haplotype, whereas most of the other inbred strains of mice carry either CD72^a or CD72^b. In this study, we generated congenic NOD.CD72^b mice that carry C57BL/6 (B6) mouse-derived centromeric chromosome 4 interval (24-45 cM) surrounding the CD72^b locus. Unexpectedly, NOD.CD72^b mice were not protected from diabetes, but rather exhibited accelerated development of both insulitis and diabetes. Our result defines novel locus or loci in the vicinity of CD72 gene that negatively control diabetes, indicating that NOD disease is under complex genetic controls of not only Idd genes but also disease-resistant genes.     

Functional interaction between the ubiquitin-specific protease 25 and the SYK tyrosine kinase

The SYK non-receptor tyrosine kinase is a key effector of immune receptors signaling in hematopoietic cells. Here, we identified and characterized a novel interaction between SYK and the ubiquitin-specific protease 25 (USP25). We report that the second SH2 domain of SYK physically interacts with a tyrosine-rich, C-terminal region of USP25 independently of tyrosine phosphorylation. Moreover, we showed that SYK specifically phosphorylates USP25 and alters its cellular levels. This study thus uncovers a new SYK substrate and reveals a novel SYK function, namely the regulation of USP25 cellular levels.     

The pre-B cell receptor checkpoint

B lymphocytes are essential antibody-producing cells of the immune system. During the development of progenitor B cells to mature B cells that express a membrane-bound antibody, the B cell receptor (BCR), the cells undergo selection at several checkpoints, which ensures that a diverse antibody repertoire is generated and that the BCRs recognise foreign-, but not self-, antigens. In this review, we consider the pre-BCR checkpoint. Mutations or alterations that affect this checkpoint underpin the development of pre-B cell leukemias, primary immunodeficiency, and possibly, systemic autoimmunity.