Biomedical subjects

Crispin R Dass

Publications and source records attributed to Crispin R Dass.

At least 19 recordsLinked to original sources

PubMed2006

A novel orthotopic murine model provides insights into cellular and molecular characteristics contributing to human osteosarcoma.

As a reliable model for osteosarcoma is lacking, three human cell lines (SaOS-2, U2OS and 143B) were evaluated in cell-based assays for proliferation, adhesion, migration, invasion, anchorage-independent growth, angiogenesis, mineralised nodule formation, plasmid transfection and oligonucleotide transfection. Tumor take and metastasis after orthotopic injection of the three cell lines into mice was monitored. The levels of expression of typical bone markers were determined with semi-quantitative RT-PCR in cultured cells, primary tumors, and for the SaOS-2 cell line, the metastases. Tumors grew and spread to the lungs within 3 and 5 weeks respectively, mimicking the clinical progression of the disease as analysed by x-ray. Expression of molecular markers in SaOS-2 indicated a mostly differentiated cell type at the primary and secondary sites. The ability of osteosarcoma cells to interact with collagen-1 and to form mineralised deposits correlated positively with tumor aggression in vivo. Expression of alkaline phosphatase was a common theme in both tumor models at the primary site. The newly established SaOS-2 model should allow the testing of candidate anti-osteosarcoma agents as well as dissection of more intricate mechanisms involved in human osteosarcoma.

Alkaline Phosphatase↗

PubMed2006

The urokinase plasminogen activator receptor as a gene therapy target for cancer.

Urokinase plasminogen activator (uPA) and/or its receptor (uPAR) are essential for metastasis, and overexpression of these molecules is strongly correlated with poor prognosis in a variety of malignant tumours. Impairment of uPA and/or uPAR function, or inhibition of the expression of these components, impedes the metastatic potential of many tumours. Several approaches have been employed to target uPAR with the aim of disrupting its ligand-independent action or interaction with uPA, including the more recent antigene technology. This review discusses the in vivo use of antigene approaches for downregulating uPAR as a potential therapy for cancer. Preclinical studies are advancing towards the translational phase, provided that established orthotopic tumours, which mimic clinical progression and presentation, are treated using clinically acceptable modes of nucleic acid delivery.

Animals↗

PubMed2006

Human xenograft osteosarcoma models with spontaneous metastasis in mice: clinical relevance and applicability for drug testing.

Osteosarcoma cells derived from patients have been isolated and subsequently cultured for the past 35 years. To date though, there have been no major breakthroughs in the development of a model for osteosarcoma that uses orthotopic implantation of human osteosarcoma cells and that closely emulates the clinical progression of this debilitating and fatal disease. Such a model is long overdue given the devastating demographics (second highest cause of cancer-related death in the paediatric age group) of the ailment and the lack of solid options for control, if not cure, for the disease, as it also is the most common primary tumour of bone. Only then can more robust R & D be undertaken in the search for efficacious anti-osteosarcoma agents. This review tackles this conundrum and lists the variety of models (that use human osteosarcoma cells) available and the types of studies performed with these.

Adolescent↗

PubMed2006

Biophysical delivery of peptides: applicability for cancer therapy.

There is a current trend towards evaluation of molecular agents for treatment of a variety of ailments, including cancer. One class of such biomolecules is proteins, and their shortened versions, peptides. Use of peptidic entities has been hindered by poor bioavailability in vivo and the high cost involved in mass-producing these macromolecular drugs. The need for localized delivery is being met with the development of various biophysical means, which include devices and aids, mainly transdermal and invasive implants. In addition, various cell-based delivery modalities, which include the use of spore-forming bacteria and stem cells, are being explored. This review discusses these methods in turn, and examines ways by which these can be enhanced for peptide delivery to tumors.

Animals↗

PubMed2006

PEDF: a potential molecular therapeutic target with multiple anti-cancer activities.

Pigment epithelium-derived factor (PEDF) is an endogenously produced protein that is widely expressed throughout the human body, and exhibits multiple and varied biological activities. Already established as a potent anti-angiogenic molecule, PEDF has recently shown promise as a potential anti-tumour agent, causing both direct and indirect tumour suppression. Here, we explore the unique anti-tumour properties of PEDF and discuss its role as an effective anti-angiogenic, anti-proliferative and pro-differentiation factor. We also discuss the prospects for PEDF therapy and the need for a closer evaluation of issues such as delivery, stability and potential toxicity.

Angiogenesis Inhibitors↗

PubMed2006

Commonly used mouse models of osteosarcoma.

Osteosarcoma is the commonest primary tumour of bone and the second highest cause of cancer-related death in the paediatric age group. Little is known of the aetiology of human osteosarcoma and lesser still of the various interactions that occur between host and tumour cells that govern growth and progression of osteosarcoma in vivo. Although numerous osteosarcoma cell lines have been established and characterized in vitro, some as far back as in the 1960s, there is a scarcity of reliable and reproducible in vivo animal models that mimics all aspects of the human condition at the temporal, physiological and histopathological level, hence, making the accurate testing of therapeutic strategies difficult. Given that osteosarcoma is a disease that affects young people and better disease management strategies are essential, development of a robust human osteosarcoma model is long overdue.

Animals↗

PubMed2006

Targeting of small molecule anticancer drugs to the tumour and its vasculature using cationic liposomes: lessons from gene therapy.

Cationic (positively charged) liposomes have been tested in various gene therapy clinical trials for neoplastic and other diseases. They have demonstrated selectivity for tumour vascular endothelial cells raising hopes for both antiangiogenic and antivascular therapies. They are also capable of being selectively delivered to the lungs and liver when administered intravenously. These vesicles are being targeted to the tumour in various parts of the body by using advanced liposomal systems such as ligand-receptor and antibody-antigen combinations. At present, the transferrin receptor is commonly used for cancer-targeted drug delivery systems including cationic liposomes. This review looks at the growing utility of these vesicles for delivery of small molecule anticancer drugs.

Journal Article↗

PubMed2006

Carrier-mediated delivery of peptidic drugs for cancer therapy.

Protein and peptide drugs are used for treatment of a variety of ailments. However, their wider use has been hindered by issues such as poor bioavailability in vivo and the cost involved in producing these drugs. This review discusses the various carrier-mediated methods used for delivery of peptide and protein drugs, with emphasis on liposomal and microspherical drug delivery systems. A brief look at the types of peptidic drugs currently in use clinically, and a brief discourse on several novel ideas for better protein delivery systems for cancer therapy is included.

Drug Carriers↗

PubMed2006

Selective gene delivery for cancer therapy using cationic liposomes: in vivo proof of applicability.

Targeted gene therapy is essential if cancer treatment is to become a reality with this form of therapy. In the past few years, cationic liposomes, discovered 2 decades ago, and at present, the most commonly used class of transfection reagents, have been tested in various clinical trials for diseases not restricted to cancer. They have been shown to be selective for tumour vascular endothelial cells raising hopes for antiangiogenic and antivascular therapies. They are also capable of being selectively delivered to the lungs and liver when administered intravenously. These vesicles are also being targeted to the tumour in various parts of the body by using advanced liposomal systems such as antibody-antigen and ligand-receptor combinations. This review looks at the state of play in this rapidly growing field.

Animals↗

PubMed2006

Downregulation of uPAR confirms link in growth and metastasis of osteosarcoma.

The uPA/uPAR system is involved in tumour progression and metastasis of a variety of cancers. Previously, we have shown that increased expression of urokinase plasminogen activator (uPA) correlated with malignancy grade in certain sarcomas. A study looking at in vivo inhibition of this system has not been done to date for osteosarcoma. More recently, this laboratory developed a clinically relevant mouse model where intratibial injection of UMR106-01 cells resulted in the development of osteosarcoma and lung metastases. Expression of uPA and its receptor (uPAR) were localised to the invading front of the tumours. Pulmonary metastasis is a predominant feature of the disease and is the major cause of death in patients. In the present study, the effects of down-regulating uPAR were observed in vitro and in vivo. UMR106-01 cells were transfected with either antisense-uPAR or vector control plasmids. Two antisense clones, exhibiting uPAR downregulation, demonstrated decreased adhesion, migration and invasion in cell-based assays in vitro (P<0.05). Cellular proliferation was not affected by uPAR downregulation. In vivo, a marked reduction of 80% in tibial tumour volumes (P<0.05), and total inhibition of pulmonary metastases were observed in mice injected with the more potent of the antisense clones. This study proves seminally the usefulness of uPAR antisense in curbing the growth and spread of osteosarcoma.

Animals↗

PubMed2006

Preclinical anticancer activity of DNA-based cleavage molecules.

Deoxyribozymes (DNAzymes) are DNA residue-based molecules capable of specific cleavage of complementary mRNA. As such, they are more stable counterparts for the earlier discovered ribozymes. A handful of studies have shown the potential of DNAzymes against cancer both in cell culture and importantly in vivo models. This relatively new molecular entity may progress to clinical trials provided that more extensive testing is carried out at the preclinical stage. While a significant amount of work has gone into chemically stabilizing the molecule, delivery is one area that needs particular attention.

Animals↗

PubMed2006

Pigment epithelium-derived factor: a multimodal tumor inhibitor.

Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serine protease inhibitor (serpin) family, is a well-known potent endogenous inhibitor of angiogenesis. It has been known for years to be aberrantly expressed in ocular disorders, but in recent years, down-regulation has been shown to be prevalent in a range of cancers as well. This review describes the trimodal anticancer activities of this interesting protein: antiangiogenesis, apoptosis-mediated tumor suppression, and tumor cell differentiation. The key to successful antitumor therapy with this protein is the ability to synthesize the recombinant form of the protein (or its active shortened forms) and deliver at therapeutic doses or alternatively to use gene transfer technology to prolong the effect in vivo. Although there is a substantial amount of work carried out at the preclinical stage with this protein, more groundwork has to be done before PEDF is tested against cancer in clinical trials.

Angiogenesis Inhibitors↗

PubMed2006

In-vitro and in-vivo assays for angiogenesis-modulating drug discovery and development.

In the past 35 years, significant findings have been made in relation to angiogenesis, and how this usually normal physiological function is converted into an abnormal state in cancer. To search for agents that can inhibit angiogenesis, and thereby prevent a tumour from proliferation and spread that is ultimately fatal to the patient, various in-vitro assays have been developed. In addition, older assays have been refined usually into high throughput screening formats, mainly by the biopharmaceutical industry in their attempts to develop novel therapeutic molecules and maintain a pipeline of lead candidates. The central aim is to extract more accurate data that would facilitate the birth of innovative mechanisms to defeat aberrant angiogenesis in-vivo. At the same time, better in-vivo models have been established, with the goal to mimic as close as possible the natural progression of various types of neoplasms in response to a good angiogenic response. More clinically relevant models are needed as anti-angiogenesis drug discovery and drug development companies fast track their lead molecules from preclinical investigations to phase I clinical trials.

Angiogenesis Inhibitors↗

PubMed2004

Lipoplex-mediated delivery of nucleic acids: factors affecting in vivo transfection.

For the past 15 years cationic liposomes have routinely been utilised for the delivery of nucleic acids such as plasmids and oligodeoxynucleotides to cells in culture and in vivo. These reagents are commercially available or are formulated inhouse. However, particularly in cultured cells, toxicity remains a significant problem, and this is confirmed by several in vivo findings. In addition, these complexes exhibit an immunostimulation effect, a phenomenon that may either be harmful or beneficial. Furthermore, lipoplexes have been recently found to have a certain degree of selectivity for dividing vascular endothelial cells. The development of cationic lipids that are safe to use especially in vivo and possess enhanced transfection capabilities is an ongoing process. More research is needed to understand the basic biology behind lipofection, first at the cellular level, then at the multicellular (whole organism) level.

Adjuvants, Immunologic↗

PubMed2004

Inhibition of human breast carcinoma proliferation, migration, chemoinvasion and solid tumour growth by DNAzymes targeting the zinc finger transcription factor EGR-1.

DNAzymes (synthetic catalytic DNA) have emerged as a new class of nucleic acid-based gene silencing agent. Using DNAzymes targeting the human mRNA of the immediate-early gene and C2H2-class zinc finger transcription factor early growth response-1 (EGR-1), we demonstrate here that EGR-1 plays an indispensable role in breast cancer proliferation, migration, chemoinvasion and xenograft growth in nude mice. DNAzyme inhibition of these tumorigenic processes and EGR-1 protein expression in breast carcinoma cells is sequence-specific and EGR-1 transcription-independent. These agents inhibit breast carcinoma cell migration and chemoinvasion in microchemotaxis chambers and solid tumour growth in athymic nude mice. Thus, DNAzymes targeting specific genes can inhibit multiple key tumorigenic processes in vitro and in vivo and may serve as useful anti-cancer agents.

Animals↗

PubMed2004

Effect of deoxyribozymes targeting c-Jun on solid tumor growth and angiogenesis in rodents.

BACKGROUND: The basic region-leucine zipper protein c-Jun has been linked to cell proliferation, transformation, and apoptosis. However, a direct role for c-Jun in angiogenesis has not been shown. METHODS: We used human microvascular endothelial cells (HMEC-1) transfected with a DNAzyme targeting the c-Jun mRNA (Dz13), related oligonucleotides, or vehicle in in vitro models of microvascular endothelial cell proliferation, migration, chemoinvasion, and tubule formation, a rat model of corneal neovascularization, and a mouse model of solid tumor growth and vascular endothelial growth factor (VEGF)-induced angiogenesis. All statistical tests were two-sided. RESULTS: Compared with mock-transfected cells, HMEC-1 cells transfected with Dz13 expressed less c-Jun protein and possessed lower DNA-binding activity. Dz13 blocked endothelial cell proliferation, migration, chemoinvasion, and tubule formation. Dz13 inhibited the endothelial cell expression and proteolytic activity of MMP-2, a c-Jun-dependent gene. Dz13 inhibited VEGF-induced neovascularization in the rat cornea compared with vehicle control (Dz13 versus vehicle: 4.0 neovessels versus 30.7 neovessels, difference = 26.7 neovessels; P =.004; area occupied by new blood vessels for Dz13 versus vehicle: 0.35 mm2 versus 1.52 mm2, difference = 1.17 mm2; P =.005) as well as solid melanoma growth in mice (Dz13 versus vehicle at 14 days: 108 mm3 versus 283 mm3, difference = 175 mm3; P =.006) with greatly reduced vascular density (Dz13 versus vehicle: 30% versus 100%, difference = 70%; P<.001). CONCLUSION: DNAzymes targeting c-Jun may have therapeutic potential as inhibitors of tumor angiogenesis and growth.

Angiogenesis Inhibitors↗

PubMed2004

Deoxyribozymes: cleaving a path to clinical trials.

Deoxyribozymes (DNAzymes) comprise an exciting class of nucleic acid molecules that are capable of specific cleavage of target mRNA. Recent reports attest to the potential of this class of molecules in cell culture and preclinical studies, where DNAzymes exhibit the ability to silence disease-associated genes (mainly those associated with cardiovascular disease and cancer). Rigorous testing in preclinical studies is now required before this relatively new entity enters Phase I clinical trials.

Animals↗

PubMed2004

Oligonucleotide delivery to tumours using macromolecular carriers.

Microparticles have been used for delivery of chemo- and radio-therapeutic agents to tumours in patients for more than a decade, whereas cationic liposomes have been used for introducing nucleic acids into mammalian cells for the same period of time. This minireview discusses the potential of delivering therapeutic ONs (oligonucleotides) to tumours using cationic liposomes and the major obstacles to such delivery. It also proposes how microparticles can be utilized for delivery of ONs to solid tumours. Prospects for improved ON delivery using these carriers and ways to achieve this are discussed.

Animals↗