ALDH1 as a functional marker of cancer stem and progenitor cells.

Stem cells have now been described in a variety of tissues, even in those where the cells' turn over rate is slow, such as the brain and the resting mammary gland. There is also accumulating evidence that tumors are derived from and are maintained by a rare population of dysregulated stem cells. However, discrepancies in the markers used and reported have slowed down the functional characterization of these somatic stem cells. To circumvent this challenging issue, universal stem cell markers with properties common to all stem cell types must be discovered and exploited. In line with this idea, the measurement of aldehyde dehydrogenase isoform 1 (ALDH1) activity shows promising potential as a universal marker for the identification and isolation of stem cells from multiple sources. Herein, we review the available data reporting utilization of ALDH1 activity as a means to identify and isolate stem cells and cancer stem cells, with a special focus on the mammary gland and breast cancer.

Histone tail modifications and noncanonical functions of histones: perspectives in cancer epigenetics.

Over the past few years, the histone deacetylase (HDAC) inhibitors have occupied an important place in the effort to develop novel, but less toxic, anticancer therapy. HDAC inhibitors block HDACs, which are the enzymes responsible for histone deacetylation, and therefore they modulate gene expression. The cellular effects of HDAC inhibitors include growth arrest and the induction of differentiation. Early successes in cancer therapeutics obtained using these drugs alone or in combination with other anticancer drugs emphasize the important place of posttranslational modifications of histones in cancer therapy. Histone tail modifications along with DNA methylation are the most studied epigenetic events related to cancer progression. Moreover, extranuclear functions of histones have also been described. Because HDAC inhibitors block HDACs and thereby increase histone acetylation, we propose a model wherein exogenous acetylated histones or other related acetylated proteins that are introduced into the nucleus become HDAC substrates and thereby compete with endogenous histones for HDACs. This competition may lead to the increased acetylation of the endogenous histones, as in the case of HDAC inhibitor therapy. Moreover, other mechanisms of action, such as binding to chromatin and modulating gene expression, are also possible for exogenously introduced histones.

SP analysis may be used to identify cancer stem cell populations.

Side populations (SP), as defined by Hoechst exclusion in flow cytometry, have been described a few years ago. While they represent only a small fraction of the whole cell population, their properties confer an important place in several investigations. SP cells express high levels of various members of ABC transporters family, such as MDR1 and BCRP, which are responsible for drug resistance. Targeting SP could improve cancer therapy by blocking these transporters. In addition, SP appear to be enriched in stem cells, cells that play a pivotal role in normal development and cancer biology. Thus, they could provide a useful tool and a readily accessible source for stem cell studies in both the normal and cancerous settings. However, these cells are poorly defined and pose challenges in their identification and isolation, particularly since they are few in number. Thus, better characterization of SP will advance our understanding of stem cells and will provide us an accessible target for drug resistance in cancer therapy.

Histonefection: Novel and potent non-viral gene delivery.

Protein/peptide-mediated gene delivery has recently emerged as a powerful approach in non-viral gene transfer. In previous studies, we and other groups found that histones efficiently mediate gene transfer (histonefection). Histonefection has been demonstrated to be effective with various members of the histone family. The DNA binding domains and natural nuclear localisation signal sequences make histones excellent candidates for effective gene transfer. In addition, their positive charge promotes binding to anionic molecules and helps them to overcome the negative charge of cells that is an important barrier to cellular penetration. Histonefection appears to have particular promise in cancer gene transfer and therapy.

Structure and function correlation in histone H2A peptide-mediated gene transfer.

Histone H2A has been found to be efficient in DNA delivery into a number of cell lines. We have reasoned that this DNA-delivery activity is mediated by two mechanisms: (i) electrostatically driven DNA binding and condensation by histone and (ii) nuclear import of these histone H2A.DNA polyplexes via nuclear localization signals in the protein. We have identified a 37-aa N-terminal peptide of histone H2A that is active in in vitro gene transfer. This peptide can function as a nuclear localization signal and can bind DNA. Amino acid substitutions that replace positively charged residues and/or DNA-binding residues of this peptide obliterate transfection activity. The introduction of a proline in the first turn of an alpha-helix of this 37-mer obliterates transfection activity, suggesting that the integrity of the alpha-helical structure of the N-terminal region of histone H2A is related to its transfection activity.