Normalizing the Stroma Through Targeting NG2HMP 31 mAbDirected Therapy

Antibody-based immunotherapy has been greatly investigated as a therapeutic strategy against NG2 HMP-expressing melanomas. This method utilizes a molecular vehicle, mAb, to selectively deliver radionuclides or toxins to tumor cells (100). Several studies have used various mAbs that recognize the NG2 HMP because the majority of melanomas extensively express it. There is limited intra- and intertumor heterogeneity, where it has been detected on > 90 of melanoma cells (49,101,102). Furthermore,...

Preclinical Development and Validation of Liposome Entrapped raf Antisense Oligonucleotide

A schematic diagram of LErafAON formulation is shown in Fig. 4A. The rafAON is a 15-mer ASO with phosphorothioate linkage limited to one base at 5'- and 3'-ends (30). The rafAON sequence is targeted against the translation initiation region of c-raf-1 mRNA. The liposome formulation consists of a mixture of a cationic lipid (dimethyldioctadecyl ammonium bromide), egg phos-phatidylchloline, and cholesterol in a molar ratio of 1 3.2 1.6. The rafAON to lipid ratio is 1 15 (w w). The particle size...

Andreas Herbst and Frank Thomas Kolligs

The Wnt p-catenin signaling pathway is tightly regulated and has important functions in development, tissue homeostasis, and regeneration. Deregulation of Wnt p-catenin signaling is frequently found in various human cancers. Eighty percent of colorectal cancers alone reveal activation of this pathway by either inactivation of the tumor-suppressor gene adenomatous polyposis coli or mutation of the proto-oncogene P-catenin. Activation of Wnt p-catenin signaling has been found to be important for...

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Regression of colorectal polyps in humans colon cells lacking Wnt-1 expression were not affected by the antibody treatment. The same group also analyzed the expression of the Wnt antagonist WIF-1 in colorectal cancer cell lines (156). WIF-1 expression was found in normal colon cells, but was missing or downregulated in the cancer cells. The reduced expression of WIF-1 was found to be caused by hypermethyla-tion of the WIF-1 promoter. Restoration of WIF-1 expression, silencing of Wnt-1 by siRNA,...

Hydrodynamic Intravascular Injection

The large volume hydrodynamic delivery technique was a major advance in the in vivo delivery of nucleic acids into mice and rats. In principle, it means the injection of a high volume into the vascular system within a short period of time. The first report that this delivery method can be even more effective than direct injection into the target organ is from Budker et al. (13), who injected high volumes of either lacZ or luciferase reporter plasmids into the portal vein of mice. Using this...

CSF1 Antisense Treatment Suppresses Growth of Human Embryonic and Colon Carcinoma Xenografts in Mice

Human Embryonic Cancer Cells Upregulate Host CSF-1 Production Human embryonic cancer cells (CRL-2073) show no detectable mRNA or protein for human CSF-1 or CSF-1R in vitro. When these cells are xenografted into the testis of SCID mice, however, mouse tissue CSF-1 gene and protein expression increases significantly compared to untreated mice. Associated with increasing CSF-1 tissue expression is an enhanced infiltration of macrophages within and surrounding the tumor. These findings...

The Brain Stroma as the Glioma Backdrop

The human brain is made up of numerous cell types that interact physically through cell-cell, and cell-extracellular matrix (ECM) contacts, and biochemically via soluble and insoluble signaling molecules. Neuroepithelial cells of the central nervous system (CNS) constitute neuroglia (astrocytes, oligodendro-cytes, and microglia) and neurons, which are derived from the neuroectoderm. These cells produce the ECM, an important feature of the normal stroma, which provides structural scaffolding as...

Influence of Genetic and Other Factors on Drug Effects in Cancers

Cancer progression is characterized by the accumulation of multiple genetic mutations, chromosomal instabilities, and or epigenetic changes that cooperate to drive malignancy (7). Also, changes in gene expression are further affected by the microenvironment. Indeed, it is now well established that the microenvironment of the tumor-host interface plays a proactive role during malignant disease progression, including the transition from carcinoma in situ to invasive cancer, tumor cell...

Heterotrimeric G Proteins and Disease 0yvind Melien

Heterotrimeric G proteins attached to the cell membrane convey signals from G proteincoupled receptors in response to stimulation by a number of hormones, neurotransmitters, chemokines, and pharmacological agents to intracellular signaling cascades. The heterotrimeric G proteins are also located in the cell interior, and receptor-independent mechanisms may elicit their activation. Thus, G proteins may possibly exert cellular functions other than acting as signaling transducers. There is also...

NG2 at the Tumor Stroma Interface

NG2 is a 300-kDa membrane spanning CSPG that was originally identified as a surface epitope on a subset of tumor cell lines derived from rat embryos after ethylnitrosourea administration (6). The cell lines were characterized by the presence of Na+ and K+ ion channels, but an inability to generate full-fledged action potentials, features characteristic of neural precursor cells. NG2 positive neural precursors can differentiate into either neuroglia or neurons, hence the designation nerve glial...

Expression wtLfLHttf

Anomalies) is transmitted as an autosomal recessive trait. It is characterized by immune deficiency in association with unstable paracentromeric heterochromatin instability (extensively related with hypomethylated genomic regions) and facial dysmorphism. Patients are affected by recurrent respiratory infections beginning in childhood. The syndrome directly results from mutations in the gene encoding for DNA-methyltransferase 3B. This may explain the hypomethylation in the pericentromeric...

Mouldy Sioud and 0yvind Melien

Although several drug targets are identified, current strategies in therapy do not take into account that patients vary in their response to drugs, both with respect to efficacy and toxic side effects. Whereas both clinical and histopathologic predictors of prognosis are established in some diseases, a better understanding of the molecular mechanisms that determine treatment response should play an important role in the development of individualized medicine. Treatment optimization will rely on...

Therapeutical Use of siRNA to Prevent and Treat Acute Liver Failure in Mice

As the first demonstrations of a therapeutic application of siRNA in vivo were done in mouse models of acute liver failure (ALF), the next paragraphs will give a brief introduction into this disease, followed by a discussion of the therapeutic use of siRNA to suppress apoptosis in this model. ALF is defined as a dramatic clinical syndrome in which a previously normal liver fails within days or weeks. Three subgroups of ALF can be distinguished, hyperacute, acute, and subacute liver failure....

DNA Methylation and Histone Modifications in Patients With Cancer

Potential Prognostic and Therapeutic Targets Michel Herranz and Manel Esteller Epigenetics, a combination of DNA modifications, chromatin organization, and variations in its associated proteins, configure a new entity that regulates gene expression throughout methylation, acetylation, and chromatin remodeling. In addition to silencing as a result of mutations, loss of heterozygosity, or classical genetic events epigenetic modification symbolizes essential early events during carcinogenesis and...

CSF1 in Breast Cancer

The mechanism by which mammary epithelial cells undergo genetic changes that result in acquisition of the ability to invade and colonize distant sites is complex (2,41,42). Normal and malignant mammary epithelium and the surrounding stromal cells produce and respond to various growth factors. Among the stromal cells, macrophages play a unique role because they are recruited into mammary gland carcinomas (43,44). The fact that in the absence of such tumor-associated macrophages, metastatic...

Hydrodynamic High Volume Tail Vein Injection in Mice see Note

In principle, any mouse strain can be used for this technique. BALB c or other strains with white fur are preferred as visualization and puncture of the tail vein, especially with bigger needles, is more difficult in black mice. We prefer to perform standard hydrodynamic tail-vein injection without anesthesia, as the combination of the high-volume injection together with anesthesia can lead to complications in some mice. If anesthesia is used, a gentle gas-anesthesia (e.g., isoflurane) should...

Lars Zender and Stefan Kubicka

RNA interference (RNAi) is a sequence-specific gene-silencing mechanism triggered by double-stranded RNA. RNAi was shown to allow transient or stable knockdown of gene expression in a broad range of species and has been used successfully for functional genomic screens in mammalian cells and Caenorhabditis elegans. Standard therapeutic use of RNAi in clinical settings in humans has been hampered by the lack of effective methods to deliver the small-interfering RNAs (siRNAs) or short-hairpin RNA...

In Vivo Delivery of Antisense Oligonucleotides

The criteria of a clinically and commercially compatible ASO delivery system include nanoparticle size (< 1 in diameter), biodegradability, high encapsulation entrapment efficiency, ASO stability and favorable pharmaco-kinetics, tumor-specific drug delivery, easy formulation, scalability, and safety profiles in animals and humans. Such an optimal in vivo delivery system offers merits that may complement not only the chemically modified ASO but also natural ASO. First, a suitable vehicle may...

Druggable Signaling Proteins

In normal cells, signaling pathways are tightly regulated. However, when they are aberrantly activated, certain pathways are capable of causing diseases. In many tumors, the aberrantly activated signaling proteins include members of the epidermal growth factor receptor family, the Ras proteins, protein kinase C isoenzymes, BCR-ABL fusion protein as well as transcription factors such as signal transducers and activators of transcriptions and Myc. Accordingly, deregulation of these signaling...

AntiCD40 Ligand Therapy

Production of pathogenic antibodies in SLE requires T-cell help along with ligation of the B-cell surface immunoglobulin receptor by antigen. It is likely that macrophages, dendritic cells, and endothelial cells are also activated by interaction with T-cells and contribute to lupus pathology. CD40 ligand (CD40L, CD154), a member of TNF family of cell surface molecules, mediates these contact-dependent signals delivered by CD4+ T helper cells to CD40+ target cells (46). Several tissue injuries...

Efficient Delivery the Holy Grail of Gene Therapeutic Approaches

The holy grail of all in vivo gene therapeutic approaches is to deliver effective amounts of the therapeutic gene into the desired target cells. Currently, viral vectors are most widely used for delivery of therapeutic genes in vivo. However, vector-associated side effects limit their value as a tool for clinical use. Systemic application of plasmid DNA or antisense oligonucleotides ASO , however, leads to very low uptake in most tissues. In part, this is owing to the fact that nucleic acids...

Martha Chekenya and Heike Immervoll

Neuroepithelial cells of the central nervous system constitute neuroglia astrocytes, oligodendrocytes, and microglia , ependyma, and neurons, which make up the stromal cells of the brain. The stromal tissue organization of the brain is tightly regulated, but occasionally the signals that define the normal contexts become disrupted and result in cancer. Malignant progression is then maintained by cross-talks between the tumor and its stroma, where the activated stroma nurtures the proliferative...