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PubMed1992

Immunotherapy with cytokine gene-transduced tumor cells: the next wave in gene therapy for cancer.

Recently, new tumor vaccine approaches were developed in animal systems that modify tumor cells genetically to secrete certain cytokines. Engineering tumor cells to secrete cytokines in a paracrine fashion can induce powerful local cytokine effects without producing significant systemic toxicity. In addition to local inflammation, this approach can alter the presentation of tumor antigens or activation of tumor antigen-specific T lymphocytes, resulting in systemic antitumor immunity. The development of high efficiency gene transfer technologies such as defective retroviral vectors allows for the translation of these preclinical studies to clinical trials. However, before large investments are made in this area of gene therapy, it will be important to demonstrate that the actual gene transfer component of the strategy significantly enhances antitumor immune responses relative to alternative nongenetic approaches.

Cytokines↗

PubMed1988

The choice of prosthetic and orthotic technique for less developed countries: analysis and perspectives in Colombia.

In their efforts to set up orthopaedic technical services, less developed countries such as Colombia have often relied on so called "turn key" technology transfers depending on the know-how and the infrastructures in industrial countries. The sophisticated industrial product stands thus in sharp contrast to the "single item" product which relies on traditional crafts and local materials. Both extreme forms of technology are currently employed in most less developed countries including Colombia. While the high-technology product is costly and requires a specialized base to produce and to service it, the "single item" product lacks generally the minimum requirements in terms of uniform quality, biomechanical function and cosmetic acceptability. Although the remarkably high standard of industrial products explain their worldwide demand and distribution, it should not be concluded that countries such as Colombia have no other choice than to adopt these technologies. In search of an intermediate alternative, the potential of a yet undervalued, appropriate technology is explored and an accurate account of its advantages is given.

Colombia↗

PubMed1997

Gene delivery to the epidermis.

Epidermal gene delivery techniques are being developed as an experimental approach to understanding the pathogenesis of skin disorders and for developing therapeutic strategies for the treatment of disease. This technology is being evaluated in many clinical trials in the treatment of disorders such as cutaneous melanoma and skin wounding, with 20% of all gene therapy protocols being applied in the field of dermatology. This review focuses on recent advances in the development of gene transfer technology to the epidermis, describing the diseases that may be amenable to treatment by use of these strategies. We will discuss the advantages and limitations of the currently used techniques and the future prospects for gene therapy via the epidermis.

Animals↗

PubMed1988

Use of transgenic animals to study human retroviruses.

The purpose of transgenic technology is to introduce a gene into the germline of an animal in order to investigate its proper expression in the appropriate cell types and its effect on cellular functions. This technology has been used to explore a wide variety of genetic and biological issues including stage- and tissue-specific gene expression, development, immunology, oncology, and gene therapy. The intent of this article is to discuss the value of gene transfer technology for the study of mechanisms of viral pathogenesis. We will focus mainly on our experience with two experimental systems involving human T-lymphotropic virus Type 1 (HTLV-1) and human immunodeficiency virus Type 1 (HIV-1).

Animals↗

PubMed2000

A study of the inheritance of a bleeding disorder in Simmental cattle.

A study was designed to determine the inheritance pattern of a blood platelet aggregation disorder in Simmental cattle utilizing embryo transfer technology. A Simmental donor cow that had previously produced a calf with the platelet aggregation disorder was superovulated and mated to a bull that had also produced affected offspring. Twenty-seven calves were produced from the 63 (42.9%) embryos transferred. This somewhat lower than expected pregnancy rate is suggestive of an increased rate of embryo loss. Twenty-three of 25 (92%) calves had normal platelet aggregation patterns and 2 failed to show any evidence of platelet aggregation. Data are suggestive that inheritance is not simple Mendelian recessive. A more likely scenario is that the defect is the result of the inheritance of at least 2 genes, which is also consistent with the sporadic incidence reported in the population at large.

Animals↗

PubMed1996

Retroviral vectors. From laboratory tools to molecular medicine.

The majority of clinical trials for gene therapy currently employ retroviral-mediated gene delivery. This is because the life cycle of the retrovirus is well understood and can be effectively manipulated to generate vectors that can be efficiently and safely packaged. Here, we review the molecular technology behind the generation of recombinant retroviral vectors. We also highlight the problems associated with the use of these viruses as gene therapy vehicles and discuss future developments that will be necessary to maintain retroviral vectors at the forefront of gene transfer technology.

Animals↗

PubMed2004

Tropical diseases, pathogens, and vectors biodiversity in developing countries: need for development of genomics and bioinformatics approaches.

The world's biodiversity, including many infectious, parasitic disease agents and their vectors whose impact on both human and animal health is significant, is largely retained in the developing countries of the tropics. Owing to the number of species involved and the relatively low-level exploration of pathogens and vectors biodiversity, several organisms are still waiting to be discovered and consequently explored in terms of genomics. Although some parasitic species of humans and animals have been studied through genomics and bioinformatics approaches, a significant number of relevant species are still to be addressed. Through the use of modern technologies, such as genomics and bioinformatics, for assessment of biodiversity and targeting tropical diseases, other relevant advantages of these initiatives for developing countries would be technology transfer and capacity building. Consequently, these initiatives could be critical to the development of the respective countries. Moreover, intra- and interhemispheric scientific collaboration should be encouraged and supported to increase the chances for success. In Brazil, the Ministry of Science and Technology has stepped forward to further such initiatives, co-supporting collaborative genomics and bioinformatics projects. The need for the establishment of working groups on genomics and bioinformatics in developing countries as well as the improvement and strengthening of collaborative research projects between developed and developing countries is discussed from our point of view. As these discussions remain open to debate, we encourage colleagues to promote further discussion on the subject.

Animal Diseases↗

PubMed1992

Structure of breeding programs to capitalize on reproductive technology for genetic improvement.

Potential rates of genetic progress are limited by biological constraints, which along with genetic parameters determine the structure of breeding programs to be employed for maximum genetic improvement. The objective here is to determine whether current progeny-testing programs in dairy cattle, which have been dictated and constrained by low female reproductive rates, need to be changed to capitalize on new reproductive technologies and how these changes should be implemented. Many differences between breeding programs diminish when selection on animal model genetic evaluations across all age and population groups is adopted as a strategy. Progeny-testing schemes then evolve toward dispersed open nucleus breeding schemes when multiple ovulation and embryo transfer is used on bull-dams. Nucleus breeding schemes have been advocated to capitalize on embryo transfer technology. In nucleus breeding schemes utilizing high reproductive rates, inbreeding, rather than reproductive rate, poses a limit to genetic progress, and strategies that maximize response to selection while limiting inbreeding need to be employed. One strategy is mating each dam to several sires rather than only one sire. In vitro embryo production techniques can be used to facilitate such mating strategies. Large-scale in vitro embryo production programs, in which large numbers of embryos per female are tested in the commercial population, offer the greatest potential for genetic gain with low rates of inbreeding. Cloning has an impact mainly on methods for dissemination of genetic improvement. Breeding herds, genetically inferior to marketed clones, are needed for continuous genetic gain. Reproductive technologies offer the potential for genetic improvement. Whether new breeding programs require changes in population structure, e.g., by creation of nucleus breeding herds, depends mainly on logistics and on quantity and quality of field information.

Animals↗

PubMed2004

Clinical gene transfer studies for hemophilia A.

The recent advances in gene transfer technology have expedited the development of gene therapy for the treatment of hemophilia A. Three different U.S. Food and Drug Administration-approved phase I clinical trials had been initiated using different gene therapy approaches each with their own advantages and limitations. In the first gene therapy trial for hemophilia A, a non-viral approach was being explored for patients with severe hemophilia A using ex vivo transfected dermal fibroblast expressing B-domain-deleted factor VIII ( BDD-FVIII). There were no serious adverse events and some patients appeared to have experienced fewer bleeding episodes with very low levels of FVIII near baseline. In the second trial, onco-retroviral vectors expressing BDD-FVIII were injected by peripheral intravenous infusion in adult patients suffering from severe hemophilia A. The procedure was safe and in some patients FVIII-transduced cells were detectable in the peripheral blood for more than a year. Although no sustained FVIII expression was detectable, occasional modest changes in FVIII levels were apparent, and in some cases a reduced bleeding frequency occurred compared with historical rates. In another trial, one patient suffering from severe hemophilia A has been treated with a high-capacity (or gutless) adenoviral vector expressing full-length FVIII, which appeared to have resulted in 1% of normal FVIII levels for several months. However, a transient inflammatory response with hematologic and liver abnormalities was observed. In conclusion, although modest improvements in clinical end points have been detected in some patients in these early phase I trials, further improvements in gene delivery technologies are warranted to bring hemophilia A gene therapy one step closer to reality.

Clinical Trials as Topic↗

PubMed2004

Four-dimensional multiphoton confocal microscopy: the new frontier in cellular imaging.

This paper reviews new developments in microscopy that combine gene transfer technology, multiphoton confocal fluorescence microscopy, live cell imaging and digital imaging techniques that provide unique insights into the complex physiological processes involved in tissue function at the cellular and subcellular level. The evolution of this novel, new technology is discussed with particular attention to earlier achievements in noninvasive ocular surface imaging. The practical basis of confocal microscopy, multiphoton confocal fluorescence microscopy, and the vital fluorescent labeling of cells in living tissues are also discussed. Additionally, one application using retroviral gene transfer to express enhanced green fluorescent protein in living wound healing fibroblasts is presented as an example of how living biology can be studied in situ in four dimensions (x, y, z, time).

Journal Article↗

PubMed1990

Multiple superovulations in N'Dama heifers.

Five N'Dama heifers were superovulated with follicle stimulating hormone (FSH-P or Folltropin) a total of six times each. The superovulations were carried out between ongoing experimental Trypanosoma congolense infections. Twenty-four (80%) of the 30 superovulations had a good ovarian response with 21 (70%) producing an average of 2.7 +/- 0.4 (mean +/- s.e.m.) embryos. The highest embryo production was achieved at the third and fourth superovulation, after which both the number of embryos and their quality declined. The overall pregnancy rate after transfer into Boran (Bos indicus) cow recipients was 50.9%. The uteri of the heifers increased considerably in size throughout the six superovulations which made it difficult to flush some of the animals after the third superovulation. Embryo transfer technology is a useful breeding tool in N'Dama heifers and multiple superovulations can be carried out with success.

Animals↗

PubMedDate not supplied

Intratumoral gene therapy for non-small cell lung cancer: current status and future directions.

Major advances in the field of molecular genetics and cancer biology have allowed for the development of rationally designed anti-cancer strategies. When genetic material (DNA or RNA) is used for this purpose, this approach is called gene therapy. Cancer cells are remarkably sensitive to the inhibition of activated oncogenes or replacement of lost tumor suppressor gene (TSG) function. Replacement of lost TSG p53 function has been achieved by the use of recombinant adenoviral vectors (rAdp53) that carry an expression cassette for this gene. Non-small cell lung cancer has been the subject of early clinical trials in which intratumoral injection of rAdp53 has been performed by transthoracic CT-guided injection or bronchoscopy. Gene transfer and p53 transgene expression have been documented in injected tumors as well as the safety and feasibility of this type of gene therapy. Tumor regression has been reported in rAdp53-injected lesions, either alone or in combination with systemic chemotherapy. Randomized controlled studies as well as clear demonstrations of clinical benefit are missing at present. The results of ongoing studies will allow a verdict on the future of first generation rAdp53 gene therapy. For the whole of the field of gene therapy the advent of more efficient and safe gene transfer technology will determine the speed by which this innovative therapeutic modality will meet with clinical success.

Carcinoma, Non-Small-Cell Lung↗

PubMed2003

Complementary food supplements to achieve micronutrient adequacy for infants and young children.

Many children in developing countries survive on a nutritionally inadequate diet. Dietary inadequacies during the complementary feeding period can be prevented by using complementary food supplements (CFSs) such as water dispersible or crushable micronutrient tablets, micronutrient sprinkles added to food just before feeding, or fortified spreads added to food just before feeding or fed as a snacks. A meeting was convened to discuss technical and operational issues related to the development of these new approaches and to identify knowledge gaps. The technical issues covered: what micronutrients to include, tolerable upper intake limits, bioavailability, micronutrient and macronutrient stability, package systems and amounts, encapsulation technologies, methods to limit or eliminate allergens, bacterial and chemical contamination, interactions between CFSs and complementary foods, and flavoring agents. Operational issues included: identifying the market positioning of CFSs, cost positioning of CFSs, regulatory requirements, CFS production and technology transfer, quality assurance, and public-private sector partnership and coordination. Intervention trials are needed to determine the efficacy of CFSs in preventing micronutrient deficiencies. Other important knowledge gaps relate to technical and operational issues. Sprinkles and tablets are produced using well-known technologies, but further research is needed to modify them for use as CFSs. Spread development is not as advanced as sprinkle and tablet development, and further research is needed to improve the technology. Although none of the products is ready for widespread use, enough information is available to set research priorities and accelerate product development and implementation.

Biological Availability↗

PubMed2004

Gene-based approaches for the repair of articular cartilage.

Gene transfer technology has opened novel treatment avenues toward the treatment of damaged musculoskeletal tissues, and may be particularly beneficial to articular cartilage. There is no natural repair mechanism to heal damaged or diseased cartilage. Existing pharmacologic, surgical and cell based treatments may offer temporary relief but are incapable of restoring damaged cartilage to its normal phenotype. Gene transfer provides the capability to achieve sustained, localized presentation of bioactive proteins or gene products to sites of tissue damage. A variety of cDNAs have been cloned which may be used to stimulate biological processes that could improve cartilage healing by (1) inducing mitosis and the synthesis and deposition of cartilage extracellular matrix components by chondrocytes, (2) induction of chondrogenesis by mesenchymal progenitor cells, or (3) inhibiting cellular responses to inflammatory stimuli. The challenge is to adapt this technology into a useful clinical treatment modality. Using different marker genes, the principle of gene delivery to synovium, chondrocytes and mesenchymal progenitor cells has been convincingly demonstrated. Following this, research efforts have begun to move to functional studies. This involves the identification of appropriate gene or gene combinations, incorporation of these cDNAs into appropriate vectors and delivery to specific target cells within the proper biological context to achieve a meaningful therapeutic response. Methods currently being explored range from those as simple as direct delivery of a vector to a cartilage defect, to synthesis of cartilaginous implants through gene-enhanced tissue engineering. Data from recent efficacy studies provide optimism that gene delivery can be harnessed to guide biological processes toward both accelerated and improved articular cartilage repair.

Animals↗

PubMed2004

Coordinate dual-gene transgenesis by lentiviral vectors carrying synthetic bidirectional promoters.

Transferring multiple genes into the same cell allows for the combination of genetic correction, marking, selection and conditional elimination of transduced cells or the reconstitution of multisubunit components and synergistic pathways. However, this cannot be reliably accomplished by current gene transfer technologies. Based on the finding that some cellular promoters intrinsically promote divergent transcription, we have developed synthetic bidirectional promoters that mediate coordinate transcription of two mRNAs in a ubiquitous or a tissue-specific manner. Lentiviral vectors incorporating the new promoters enabled efficient dual gene transfer in several tissues in vivo after direct delivery or transgenesis, and in a human gene therapy model. Because divergent gene pairs, likely transcribed from shared promoters, are common in the genome, the synthetic promoters that we developed may mimic a well-represented feature of transcription. Vectors incorporating these promoters should increase the power of gene function studies and expand the reach and safety of gene therapy.

Antigens, CD34↗

PubMed2005

Herpes simplex virus type 1 amplicon vectors for the infectious delivery and expression of genomic DNA loci.

A wide range of viral gene transfer technologies has been developed for gene therapy and functional genetic purposes. Different viral vectors vary in transgene capacity, range of infectivity and stability of expression. It is now becoming increasingly apparent that there are distinct advantages in using a complete genomic locus as a transgene to produce physiological levels of tissue-specific, sustained expression. However, the number of vector systems capable of delivering complete genomic loci is limited. The development of the infectious bacterial artificial chromosome system, a herpes simplex virus type 1-based vector that provides efficient infectious delivery and expression of bacterial artificial chromosome or P1-based artificial chromosome clone inserts, is reviewed. This system currently represents the leading technology capable of infectious delivery and expression of complete genomic DNA loci above 100 kb.

Chromosomes, Artificial, Bacterial↗

PubMed1998

In vivo evaluation of in vitro hypothesis using "gain or loss" of function: functional analysis of renin-angiotensin system.

Characterization of the role of components of the renin angiotensin system (RAS) in vivo is limited by the difficulty in manipulating these individual components, as well as by methodologic limitations in studying the function of a local RAS in the absence of any contribution by the circulating RAS. In vivo gene transfer technology provides us with the opportunity to study the physiologic responses to in vivo manipulation of the individual components related to hypertension (ie, by overexpression or inhibition) without changes in the circulating system. Using this technology, the importance of each component of RAS in blood pressure regulation has recently been shown. Such an approach may increase our understanding of the biology and pathobiology of the autocrine-paracrine system in hypertension. This review discusses the potential utility of in vivo gene transfer methods.

Animals↗

PubMed1994

Cellular engineering and gene therapy strategies for insulin replacement in diabetes.

In diabetes, insulin secretion is either completely absent (insulin-dependent diabetes mellitus [IDDM]) or inappropriately regulated (non-insulin-dependent diabetes mellitus [NIDDM]). In recent years, new insights into the molecular and biochemical mechanism(s) of fuel-mediated insulin release coupled with advances in gene transfer technology have led to the investigation of molecular strategies for replacement of normal insulin delivery function. Such initiatives have included attempts to engineer glucose-stimulated insulin secretion in cell lines that might serve as surrogates for islets in IDDM. The development of DNA virus gene transfer systems of remarkable efficiency also has suggested ways in which the beta-cell dysfunction of NIDDM might ultimately be repaired by gene therapy. The emerging work in these areas and implications for the future are summarized in this perspective.

Cell Line↗