Jeannette Bennicelli - Academia.edu (original) (raw)

Papers by Jeannette Bennicelli

Tenascin is a large glycoprotein of the extracellular matrix. It shows a site-restricted expressi... more Tenascin is a large glycoprotein of the extracellular matrix. It shows a site-restricted expression during embryogenesis and can be found in adult tissues during wound healing and tumorigenesis. Because of the potential involvement of tenascin in adhesion and invasion during metas tasis, the study of the interactions of tumor cells with tenascin is of considerable interest. Using five anti-melanoma monoclonal antibodies to four different epitopes of human tenascin, we found that most mela noma cells secrete tenascin in vitro constitutively. Transforming growth factor fi, in the medium increased secretion in tenascin-producing cells. Tenascin was present in sera of melanoma patients, with significantly elevated levels in patients with advanced melanomas as compared to patients with low tumor burden or to normal donors. Normal and malig nant melanocytes did not attach to tenascin as substrate within 1 to 2 h and tenascin could also inhibit fibronectin-dependent adhesion. These results...

Investigative Ophthalmology & Visual Science, 2008

L'invention concerne des compositions et des procedes pour le traitement de troubles oculaire... more L'invention concerne des compositions et des procedes pour le traitement de troubles oculaires chez un sujet. Selon un aspect, l'invention concerne un vecteur viral adeno-associe qui comprend une molecule d'acide nucleique comprenant une sequence codant pour CNGA3. Selon un autre aspect, l'invention concerne un vecteur viral adeno-associe qui comprend une molecule d'acide nucleique comprenant une sequence codant pour CNGB3. Selon un autre aspect, l'invention concerne un vecteur viral adeno-associe qui comprend une molecule d'acide nucleique comprenant une sequence codant pour REP-1. Selon des modes de realisation souhaites, le sujet est un humain, un chat, un chien, un mouton, ou un primate non humain.

Investigative Ophthalmology & Visual Science, 2005

Molecular therapy : the journal of the American Society of Gene Therapy, Jan 6, 2018

Most genetically distinct inherited retinal degenerations are primary photoreceptor degenerations... more Most genetically distinct inherited retinal degenerations are primary photoreceptor degenerations. We selected a severe early onset form of Leber congenital amaurosis (LCA), caused by mutations in the gene LCA5, in order to test the efficacy of gene augmentation therapy for a ciliopathy. The LCA5-encoded protein, Lebercilin, is essential for the trafficking of proteins and vesicles to the photoreceptor outer segment. Using the AAV serotype AAV7m8 to deliver a human LCA5 cDNA into an Lca5 null mouse model of LCA5, we show partial rescue of retinal structure and visual function. Specifically, we observed restoration of rod-and-cone-driven electroretinograms in about 25% of injected eyes, restoration of pupillary light responses in the majority of treated eyes, an ∼20-fold decrease in target luminance necessary for visually guided behavior, and improved retinal architecture following gene transfer. Using LCA5 patient-derived iPSC-RPEs, we show that delivery of the LCA5 cDNA restores le...

Molecular Therapy, 2016

Inherited retinal degenerations, such as retinitis pigmentosa (RP) and Leber congenital amaurosis... more Inherited retinal degenerations, such as retinitis pigmentosa (RP) and Leber congenital amaurosis (LCA), are characterized by progressive impairment of visual function associated with degeneration of retinal pigment epithelium (RPE) and photoreceptors. These monogenic diseases are genetically heterogeneous due to mutations in genes expressed affecting those cells. Gene therapy holds great potential for the treatment of inherited retinal diseases for which there currently is none. Efficient retinal gene transfer has been achieved with several recombinant viral vectors, including those derived from adenovirus, retrovirus, herpesvirus, and adeno-associated virus (AAV). Among these, AAV vectors appear particularly amenable to retinal gene transfer. However the success of any preclinical study depends on the availability of relevant animal models. In situations where suitable animal models are unavailable recent studies have generated proof-of concept data using personalized cell models: induced pluripotent stem cells (iPSC) derived from affected individuals and un-affected controls. With the increasing number of retinal gene therapy paradigms and recombinant vectors, in vitro bioassays characterizing vector transduction efficiency and quality are becoming increasingly important. To date, most in vitro assays using recombinant vector transduction have targeted iPSCs. To elicit additional features relevant to the disease, we differentiated iPSCs to RPE and evaluated the transduction efficiencies of a panel of AAV serotypes. Characterization of iPSC-derived RPE by qRT-PCR, immunohistochemistry, western blot analysis and flow cytometry showed the expression of typical RPE markers, phagocytic ability and gene-expression patterns similar to those of native RPE. Comparison of transduction efficiencies of different AAVs in iPSC-derived RPE was carried out using an enhanced green-fluorescent protein (eGFP) reporter gene driven by the cytomegalovirus immediate-early (CMV) promoter. At 24 to 48 hours post transduction, cells expressing GFP were identified by Typhoon scanner, and flow cytometry. Relative GFP expression was evaluated by using the software, Image-J. Of the tested AAV serotypes, AAV2 transduced iPSC-derived RPE cells most efficiently, followed by AAV7m8, AAV1 and AAV6. Differentiation into retinal neuronal cells types may require use of alternative AAV serotypes in order to obtain transduction efficiencies relevant to determination of therapeutic efficacy.

Molecular Therapy, 2015

Peroxisome biogenesis disorders (PBDs) are a group of autosomal recessive disorders most frequent... more Peroxisome biogenesis disorders (PBDs) are a group of autosomal recessive disorders most frequently caused by inherited defects in the PEX1 gene, which is required for normal peroxisome assembly and biochemical functions. Approximately 80% of patients fall within the category of Zellweger spectrum disorder (PBD-ZSD), which has an overall incidence of 1:50,000 births in the United States. The majority of patients have milder forms of disease compatible with survival through adulthood; however, they typically show mild to moderate developmental delays, and progressive vision and hearing loss. Thus, therapeutic interventions that prevent or slow down visual loss could have a profound impact on the lives of these individualsHere, we present a retinal gene therapy approach that addresses visual deterioration in patients with milder forms of disease. Optical coherence tomography (OCT) has demonstrated that the cone photoreceptor cells are most significantly affected by loss of peroxisome functions in such patients. These visual phenotypes are recapitulated in a new mouse model of disease that expresses the murine equivalent of most common PEX1 mutation found in patients (PEX1-p.G843D). Electroretinogram (ERG) analyses indicated severe impairment of the cone visual pathway in these homozygous Pex1-mutant mice by 4 months of age with the rod visual system being relatively preserved. Staining retinal sections with peanut agglutinin showed that some cone photoreceptors were retained in the homozygote murine Pex1 -mutant retina at 3 weeks, but were completely degenerated in the adult. Finally, genome-wide expression studies showed specific reduction of photoreceptor cone-specific genes with no differential expression of other cell lineage-specific genes or others that indicate cell death or stress responses.We have developed AAV vectors to deliver normal copies of the 3.85-kb human PEX1 gene to the mammalian retina. These vectors use the rhodopsin kinase 1 (RK1) or truncated cytomegalovirus (CMV) promoters, respectively. Subretinal injections of RK1.PEX1. rAAV9 in healthy mice resulted in robust expression of human PEX1 mRNA. Furthermore, transduction of cultured Pex1 -mutant murine skin fibroblasts with CMV.PEX1.rAAV9 results in the rescue of peroxisome assembly defects in these cells, as determine by the intracellular localization of a modified GFP reporter protein with a peroxisome targeting signal. This indicates that the human PEX1 transgene can complement the genetic defect in the murine Pex1 gene. We are currently testing the ability of the AAV9-PEX1 gene delivery system to complement the retinal gene defect in our Pex1 -mutant mouse model. If successful, this could provide the preliminary data required to begin initial planning for eventual clinical trials in patients.

Lancet (London, England), Jan 30, 2016

Safety and efficacy have been shown in a phase 1 dose-escalation study involving a unilateral sub... more Safety and efficacy have been shown in a phase 1 dose-escalation study involving a unilateral subretinal injection of a recombinant adeno-associated virus (AAV) vector containing the RPE65 gene (AAV2-hRPE65v2) in individuals with inherited retinal dystrophy caused by RPE65 mutations. This finding, along with the bilateral nature of the disease and intended use in treatment, prompted us to determine the safety of administration of AAV2-hRPE65v2 to the contralateral eye in patients enrolled in the phase 1 study. In this follow-on phase 1 trial, one dose of AAV2-hRPE65v2 (1·5 × 10(11) vector genomes) in a total volume of 300 μL was subretinally injected into the contralateral, previously uninjected, eyes of 11 children and adults (aged 11-46 years at second administration) with inherited retinal dystrophy caused by RPE65 mutations, 1·71-4·58 years after the initial subretinal injection. We assessed safety, immune response, retinal and visual function, functional vision, and activation ...

Scientific reports, Jan 24, 2015

The future of treating inherited and acquired genetic diseases will be defined by our ability to ... more The future of treating inherited and acquired genetic diseases will be defined by our ability to introduce transgenes into cells and restore normal physiology. Here we describe an autogenous transgene regulatory system (ARES), based on the bacterial lac repressor, and demonstrate its utility for controlling the expression of a transgene in bacteria, eukaryotic cells, and in the retina of mice. This ARES system is inducible by the small non-pharmacologic molecule, Isopropyl β-D-1-thiogalactopyranoside (IPTG) that has no off-target effects in mammals. Following subretinal injection of an adeno-associated virus (AAV) vector encoding ARES, luciferase expression can be reversibly controlled in the murine retina by oral delivery of IPTG over three induction-repression cycles. The ability to induce transgene expression repeatedly via administration of an oral inducer in vivo, suggests that this type of regulatory system holds great promise for applications in human gene therapy.

Cancer research, Jan 15, 1991

Tenascin is a large glycoprotein of the extracellular matrix. It shows a site-restricted expressi... more Tenascin is a large glycoprotein of the extracellular matrix. It shows a site-restricted expression during embryogenesis and can be found in adult tissues during wound healing and tumorigenesis. Because of the potential involvement of tenascin in adhesion and invasion during metastasis, the study of the interactions of tumor cells with tenascin is of considerable interest. Using five anti-melanoma monoclonal antibodies to four different epitopes of human tenascin, we found that most melanoma cells secrete tenascin in vitro constitutively. Transforming growth factor beta 1 in the medium increased secretion in tenascin-producing cells. Tenascin was present in sera of melanoma patients, with significantly elevated levels in patients with advanced melanomas as compared to patients with low tumor burden or to normal donors. Normal and malignant melanocytes did not attach to tenascin as substrate within 1 to 2 h and tenascin could also inhibit fibronectin-dependent adhesion. These results...

Science Translational Medicine, 2010

After successful gene therapy to correct retinal degeneration in one eye, treatment of the second... more After successful gene therapy to correct retinal degeneration in one eye, treatment of the second eye is safe and effective—even when immunity to the vector is present—in nonhuman primates and dogs.

Current Opinion in Oncology, 1999

Identification of genetic alterations has contributed greatly to the understanding of sarcoma bio... more Identification of genetic alterations has contributed greatly to the understanding of sarcoma biology. Additionally, detection of these abnormalities is providing new tools for the diagnosis of sarcomas. In this paper, three important new genetic findings from the past year are reviewed, including the t(12;15) translocation of congenital fibrosarcoma, mutation of the putative tumor suppressor gene hSNF5/INI1 in malignant rhabdoid tumor, and the association of c-kit mutations with gastrointestinal stromal tumor. Highlighted are important studies concerning mechanisms of chromosomal translocation, functions of sarcoma-specific fusion proteins, genetic abnormalities other than translocations, molecular diagnosis, and molecular profiling of gene expression. Particular emphasis is placed on information obtained with comparative genomic hybridization and microarray techniques, because these powerful technologies will facilitate the rapid acquisition of data that provide insight into the molecular genetic and biologic basis of sarcomas.

Cancer research, 1999

PAX3 and PAX7 are closely related paired box family members expressed during early neural and myo... more PAX3 and PAX7 are closely related paired box family members expressed during early neural and myogenic development. Assay of PAX3 and PAX7 mRNA expression in embryonal rhabdomyosarcoma, neuroblastoma, Ewing's sarcoma, and melanoma cell lines revealed tumor-specific expression patterns similar to the corresponding embryonic lineages. Although the mammalian PAX3 and PAX7 genes were reported to contain eight exons, we found that the predominant PAX3 and PAX7 transcripts in these tumor lines contain previously uncharacterized ninth exons. These splicing events alter the COOH-terminal coding regions of the encoded products but do not alter the transcriptional activity as assayed using a reporter gene with a model PAX3/PAX7 binding site. However, the findings of nearly identical COOH-terminal regions within the corresponding genes of the avian and fish genomes suggest conserved functional roles for these regions that require further investigation.

Cancer research, Jan 15, 1998

The 2;13 chromosomal translocation in alveolar rhabdomyosarcoma generates the chimeric protein PA... more The 2;13 chromosomal translocation in alveolar rhabdomyosarcoma generates the chimeric protein PAX3-FKHR, which is a powerful transcriptional activator. We hypothesize that PAX3-FKHR regulates downstream effector genes involved in rhabdomyosarcoma tumorigenesis. We evaluated alterations in expression of MET and neural cell adhesion molecule that were proposed previously as downstream targets of wild-type PAX3. We used a myogenic tumor cell culture system and rhabdomyosarcoma tumor specimens to assess candidate gene expression in relationship to various PAX3-FKHR expression levels. We demonstrate that the expression of MET, but not neural cell adhesion molecule, correlates significantly with PAX3-FKHR expression. These findings indicate that MET, which encodes a receptor involved in growth and motility signaling, is a downstream target of PAX3-FKHR in alveolar rhabdomyosarcoma.

Tenascin is a large glycoprotein of the extracellular matrix. It shows a site-restricted expressi... more Tenascin is a large glycoprotein of the extracellular matrix. It shows a site-restricted expression during embryogenesis and can be found in adult tissues during wound healing and tumorigenesis. Because of the potential involvement of tenascin in adhesion and invasion during metas tasis, the study of the interactions of tumor cells with tenascin is of considerable interest. Using five anti-melanoma monoclonal antibodies to four different epitopes of human tenascin, we found that most mela noma cells secrete tenascin in vitro constitutively. Transforming growth factor fi, in the medium increased secretion in tenascin-producing cells. Tenascin was present in sera of melanoma patients, with significantly elevated levels in patients with advanced melanomas as compared to patients with low tumor burden or to normal donors. Normal and malig nant melanocytes did not attach to tenascin as substrate within 1 to 2 h and tenascin could also inhibit fibronectin-dependent adhesion. These results...

Investigative Ophthalmology & Visual Science, 2008

L'invention concerne des compositions et des procedes pour le traitement de troubles oculaire... more L'invention concerne des compositions et des procedes pour le traitement de troubles oculaires chez un sujet. Selon un aspect, l'invention concerne un vecteur viral adeno-associe qui comprend une molecule d'acide nucleique comprenant une sequence codant pour CNGA3. Selon un autre aspect, l'invention concerne un vecteur viral adeno-associe qui comprend une molecule d'acide nucleique comprenant une sequence codant pour CNGB3. Selon un autre aspect, l'invention concerne un vecteur viral adeno-associe qui comprend une molecule d'acide nucleique comprenant une sequence codant pour REP-1. Selon des modes de realisation souhaites, le sujet est un humain, un chat, un chien, un mouton, ou un primate non humain.

Investigative Ophthalmology & Visual Science, 2005

Molecular therapy : the journal of the American Society of Gene Therapy, Jan 6, 2018

Most genetically distinct inherited retinal degenerations are primary photoreceptor degenerations... more Most genetically distinct inherited retinal degenerations are primary photoreceptor degenerations. We selected a severe early onset form of Leber congenital amaurosis (LCA), caused by mutations in the gene LCA5, in order to test the efficacy of gene augmentation therapy for a ciliopathy. The LCA5-encoded protein, Lebercilin, is essential for the trafficking of proteins and vesicles to the photoreceptor outer segment. Using the AAV serotype AAV7m8 to deliver a human LCA5 cDNA into an Lca5 null mouse model of LCA5, we show partial rescue of retinal structure and visual function. Specifically, we observed restoration of rod-and-cone-driven electroretinograms in about 25% of injected eyes, restoration of pupillary light responses in the majority of treated eyes, an ∼20-fold decrease in target luminance necessary for visually guided behavior, and improved retinal architecture following gene transfer. Using LCA5 patient-derived iPSC-RPEs, we show that delivery of the LCA5 cDNA restores le...

Molecular Therapy, 2016

Inherited retinal degenerations, such as retinitis pigmentosa (RP) and Leber congenital amaurosis... more Inherited retinal degenerations, such as retinitis pigmentosa (RP) and Leber congenital amaurosis (LCA), are characterized by progressive impairment of visual function associated with degeneration of retinal pigment epithelium (RPE) and photoreceptors. These monogenic diseases are genetically heterogeneous due to mutations in genes expressed affecting those cells. Gene therapy holds great potential for the treatment of inherited retinal diseases for which there currently is none. Efficient retinal gene transfer has been achieved with several recombinant viral vectors, including those derived from adenovirus, retrovirus, herpesvirus, and adeno-associated virus (AAV). Among these, AAV vectors appear particularly amenable to retinal gene transfer. However the success of any preclinical study depends on the availability of relevant animal models. In situations where suitable animal models are unavailable recent studies have generated proof-of concept data using personalized cell models: induced pluripotent stem cells (iPSC) derived from affected individuals and un-affected controls. With the increasing number of retinal gene therapy paradigms and recombinant vectors, in vitro bioassays characterizing vector transduction efficiency and quality are becoming increasingly important. To date, most in vitro assays using recombinant vector transduction have targeted iPSCs. To elicit additional features relevant to the disease, we differentiated iPSCs to RPE and evaluated the transduction efficiencies of a panel of AAV serotypes. Characterization of iPSC-derived RPE by qRT-PCR, immunohistochemistry, western blot analysis and flow cytometry showed the expression of typical RPE markers, phagocytic ability and gene-expression patterns similar to those of native RPE. Comparison of transduction efficiencies of different AAVs in iPSC-derived RPE was carried out using an enhanced green-fluorescent protein (eGFP) reporter gene driven by the cytomegalovirus immediate-early (CMV) promoter. At 24 to 48 hours post transduction, cells expressing GFP were identified by Typhoon scanner, and flow cytometry. Relative GFP expression was evaluated by using the software, Image-J. Of the tested AAV serotypes, AAV2 transduced iPSC-derived RPE cells most efficiently, followed by AAV7m8, AAV1 and AAV6. Differentiation into retinal neuronal cells types may require use of alternative AAV serotypes in order to obtain transduction efficiencies relevant to determination of therapeutic efficacy.

Molecular Therapy, 2015

Peroxisome biogenesis disorders (PBDs) are a group of autosomal recessive disorders most frequent... more Peroxisome biogenesis disorders (PBDs) are a group of autosomal recessive disorders most frequently caused by inherited defects in the PEX1 gene, which is required for normal peroxisome assembly and biochemical functions. Approximately 80% of patients fall within the category of Zellweger spectrum disorder (PBD-ZSD), which has an overall incidence of 1:50,000 births in the United States. The majority of patients have milder forms of disease compatible with survival through adulthood; however, they typically show mild to moderate developmental delays, and progressive vision and hearing loss. Thus, therapeutic interventions that prevent or slow down visual loss could have a profound impact on the lives of these individualsHere, we present a retinal gene therapy approach that addresses visual deterioration in patients with milder forms of disease. Optical coherence tomography (OCT) has demonstrated that the cone photoreceptor cells are most significantly affected by loss of peroxisome functions in such patients. These visual phenotypes are recapitulated in a new mouse model of disease that expresses the murine equivalent of most common PEX1 mutation found in patients (PEX1-p.G843D). Electroretinogram (ERG) analyses indicated severe impairment of the cone visual pathway in these homozygous Pex1-mutant mice by 4 months of age with the rod visual system being relatively preserved. Staining retinal sections with peanut agglutinin showed that some cone photoreceptors were retained in the homozygote murine Pex1 -mutant retina at 3 weeks, but were completely degenerated in the adult. Finally, genome-wide expression studies showed specific reduction of photoreceptor cone-specific genes with no differential expression of other cell lineage-specific genes or others that indicate cell death or stress responses.We have developed AAV vectors to deliver normal copies of the 3.85-kb human PEX1 gene to the mammalian retina. These vectors use the rhodopsin kinase 1 (RK1) or truncated cytomegalovirus (CMV) promoters, respectively. Subretinal injections of RK1.PEX1. rAAV9 in healthy mice resulted in robust expression of human PEX1 mRNA. Furthermore, transduction of cultured Pex1 -mutant murine skin fibroblasts with CMV.PEX1.rAAV9 results in the rescue of peroxisome assembly defects in these cells, as determine by the intracellular localization of a modified GFP reporter protein with a peroxisome targeting signal. This indicates that the human PEX1 transgene can complement the genetic defect in the murine Pex1 gene. We are currently testing the ability of the AAV9-PEX1 gene delivery system to complement the retinal gene defect in our Pex1 -mutant mouse model. If successful, this could provide the preliminary data required to begin initial planning for eventual clinical trials in patients.

Lancet (London, England), Jan 30, 2016

Safety and efficacy have been shown in a phase 1 dose-escalation study involving a unilateral sub... more Safety and efficacy have been shown in a phase 1 dose-escalation study involving a unilateral subretinal injection of a recombinant adeno-associated virus (AAV) vector containing the RPE65 gene (AAV2-hRPE65v2) in individuals with inherited retinal dystrophy caused by RPE65 mutations. This finding, along with the bilateral nature of the disease and intended use in treatment, prompted us to determine the safety of administration of AAV2-hRPE65v2 to the contralateral eye in patients enrolled in the phase 1 study. In this follow-on phase 1 trial, one dose of AAV2-hRPE65v2 (1·5 × 10(11) vector genomes) in a total volume of 300 μL was subretinally injected into the contralateral, previously uninjected, eyes of 11 children and adults (aged 11-46 years at second administration) with inherited retinal dystrophy caused by RPE65 mutations, 1·71-4·58 years after the initial subretinal injection. We assessed safety, immune response, retinal and visual function, functional vision, and activation ...

Scientific reports, Jan 24, 2015

The future of treating inherited and acquired genetic diseases will be defined by our ability to ... more The future of treating inherited and acquired genetic diseases will be defined by our ability to introduce transgenes into cells and restore normal physiology. Here we describe an autogenous transgene regulatory system (ARES), based on the bacterial lac repressor, and demonstrate its utility for controlling the expression of a transgene in bacteria, eukaryotic cells, and in the retina of mice. This ARES system is inducible by the small non-pharmacologic molecule, Isopropyl β-D-1-thiogalactopyranoside (IPTG) that has no off-target effects in mammals. Following subretinal injection of an adeno-associated virus (AAV) vector encoding ARES, luciferase expression can be reversibly controlled in the murine retina by oral delivery of IPTG over three induction-repression cycles. The ability to induce transgene expression repeatedly via administration of an oral inducer in vivo, suggests that this type of regulatory system holds great promise for applications in human gene therapy.

Cancer research, Jan 15, 1991

Tenascin is a large glycoprotein of the extracellular matrix. It shows a site-restricted expressi... more Tenascin is a large glycoprotein of the extracellular matrix. It shows a site-restricted expression during embryogenesis and can be found in adult tissues during wound healing and tumorigenesis. Because of the potential involvement of tenascin in adhesion and invasion during metastasis, the study of the interactions of tumor cells with tenascin is of considerable interest. Using five anti-melanoma monoclonal antibodies to four different epitopes of human tenascin, we found that most melanoma cells secrete tenascin in vitro constitutively. Transforming growth factor beta 1 in the medium increased secretion in tenascin-producing cells. Tenascin was present in sera of melanoma patients, with significantly elevated levels in patients with advanced melanomas as compared to patients with low tumor burden or to normal donors. Normal and malignant melanocytes did not attach to tenascin as substrate within 1 to 2 h and tenascin could also inhibit fibronectin-dependent adhesion. These results...

Science Translational Medicine, 2010

After successful gene therapy to correct retinal degeneration in one eye, treatment of the second... more After successful gene therapy to correct retinal degeneration in one eye, treatment of the second eye is safe and effective—even when immunity to the vector is present—in nonhuman primates and dogs.

Current Opinion in Oncology, 1999

Identification of genetic alterations has contributed greatly to the understanding of sarcoma bio... more Identification of genetic alterations has contributed greatly to the understanding of sarcoma biology. Additionally, detection of these abnormalities is providing new tools for the diagnosis of sarcomas. In this paper, three important new genetic findings from the past year are reviewed, including the t(12;15) translocation of congenital fibrosarcoma, mutation of the putative tumor suppressor gene hSNF5/INI1 in malignant rhabdoid tumor, and the association of c-kit mutations with gastrointestinal stromal tumor. Highlighted are important studies concerning mechanisms of chromosomal translocation, functions of sarcoma-specific fusion proteins, genetic abnormalities other than translocations, molecular diagnosis, and molecular profiling of gene expression. Particular emphasis is placed on information obtained with comparative genomic hybridization and microarray techniques, because these powerful technologies will facilitate the rapid acquisition of data that provide insight into the molecular genetic and biologic basis of sarcomas.

Cancer research, 1999

PAX3 and PAX7 are closely related paired box family members expressed during early neural and myo... more PAX3 and PAX7 are closely related paired box family members expressed during early neural and myogenic development. Assay of PAX3 and PAX7 mRNA expression in embryonal rhabdomyosarcoma, neuroblastoma, Ewing's sarcoma, and melanoma cell lines revealed tumor-specific expression patterns similar to the corresponding embryonic lineages. Although the mammalian PAX3 and PAX7 genes were reported to contain eight exons, we found that the predominant PAX3 and PAX7 transcripts in these tumor lines contain previously uncharacterized ninth exons. These splicing events alter the COOH-terminal coding regions of the encoded products but do not alter the transcriptional activity as assayed using a reporter gene with a model PAX3/PAX7 binding site. However, the findings of nearly identical COOH-terminal regions within the corresponding genes of the avian and fish genomes suggest conserved functional roles for these regions that require further investigation.

Cancer research, Jan 15, 1998

The 2;13 chromosomal translocation in alveolar rhabdomyosarcoma generates the chimeric protein PA... more The 2;13 chromosomal translocation in alveolar rhabdomyosarcoma generates the chimeric protein PAX3-FKHR, which is a powerful transcriptional activator. We hypothesize that PAX3-FKHR regulates downstream effector genes involved in rhabdomyosarcoma tumorigenesis. We evaluated alterations in expression of MET and neural cell adhesion molecule that were proposed previously as downstream targets of wild-type PAX3. We used a myogenic tumor cell culture system and rhabdomyosarcoma tumor specimens to assess candidate gene expression in relationship to various PAX3-FKHR expression levels. We demonstrate that the expression of MET, but not neural cell adhesion molecule, correlates significantly with PAX3-FKHR expression. These findings indicate that MET, which encodes a receptor involved in growth and motility signaling, is a downstream target of PAX3-FKHR in alveolar rhabdomyosarcoma.