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Archive for the ‘Gene therapy’ Category

Gene Therapy in Sheep May Bring Hope to Adults With Tay …

Thursday, August 4th, 2016

For 26 years, doctors could not piece together the medical puzzle of Stewart Altman's symptoms -- as a child growing up on Long Island, he was uncoordinated and slurred his speech. Later, as a volunteer fireman, he kept falling down and had trouble climbing the ladders.

It seemed unrelated at the time, but his older sister, who had a history of psychological symptoms, was hospitalized in a mental institution. Her psychiatrist suspected a physical disorder and consulted a geneticist who eventually connected the dots.

In 1978, Altman and his sister Roslyn Vaccaro were given a stunning diagnosis: Tay-Sachs -- an inherited neurological disease that typically affects babies, killing them between the ages of 3 and 5. Only several hundred cases exist in the United States.

Altman, now 58, has a non-fatal, adult form of the disease, late onset Tay-Sachs (LOTS), and depends on his wife and a service dog to perform most daily tasks.

"I am devastated," Altman said of the disease that has robbed him of much of his speech and muscle strength, confining him to a wheelchair. "But the alternative is much worse."

His sister died in 2000 after battling LOTS-related bipolar disorder and schizophrenia -- which occurs in 50 to 60 percent of all adult cases -- and Altman and his wife raised her two sons.

Now scientists are hopeful that gene therapy may help late-onset patients like Altman and look forward to human trials.

Tay-Sachs is caused by gene mutation results in the absence or insufficient levels of the enzyme, hexosamindase A or Hex A. Without it, a fatty substance or lipids accumulates in the cells, mostly in the brain. It comes in three forms: infantile, juvenile or adult onset.

Doctors say there can be great variations in the presentation of Tay-Sachs, even in the same family with the same mutations. Babies born with Tay-Sachs appear normal at first, but by 3 or 4 years old, their nerve cells deteriorate and they eventually die. Those with LOTS can live a long life, but, like Altman, are progressively disabled.

The story of Tay-Sachs is a miraculous one. It was first identified in the late 1800s by British ophthalmologist Warren Tay and New York neurologist Bernard Sachs, who noticed the disease was prevalent in Jews of Eastern European origin.

In the 1970s and 1980s, when genetic testing became available, synagogues launched public education campaigns encouraging prospective parents to be tested, and the disease was virtually eliminated in those of Jewish ancestry.

Now, mostly non-Jews, though their risk is not as great, are among the 100 American children who have the disease, according to the National Tay-Sachs and Allied Diseases Association (NTSAD), which leads the fight for a cure.

Altman's speech is difficult to understand, so his wife Lorrie said her husband of 37 years wanted the public to know, "it's not just an infant's disease."

"Tay-Sachs is also in the general population and people don't know," she said. "He thinks we need to get the word out. One in 250 Americans carries the gene."

French Canadians, Louisiana Cajuns and even those of English-Irish ancestry have a greater chance of carrying the recessive gene that causes the disease.

Tay-Sachs is an autosomal recessive disorder, which means each parent must carry the gene. Their children have a 25 percent chance of developing Tay-Sachs, 50 percent chance of being a carrier and a 25 percent chance of being free of that recessive gene.

Altman was born in 1952, before genetic testing was available. Both his parents were carriers of the recessive gene that causes Tay-Sachs and both he and sister were stricken with the mildest form of the disease. Two of their brothers were unaffected, although one is a carrier.

The Massapequa, N.Y., couple have two healthy sons, who are carriers, but whose wives are not, and four healthy grandchildren.

For years, Altman was able to get around with a walker until he had to drop out of a clinical trial for a new drug because of debilitating side effects. After that, he said he lost 40 pounds and so much muscle that he could no longer stand on his own.

"Between the two of us we handle it and we lead kind of a normal life," said Lorrie. "But we have no idea what the future will bring."

Altman works at Nassau University Medical Center in the security monitoring department. He raises funds for about 11 different non-profit organizations, including NTSAD, and has given presentations to the Boy Scouts and senior citizens.

Much of the public work has now ended, as his speech has become more incomprehensible because the degeneration of the nerves that control his respiratory muscles.

"Stewart has a good way of just living in the moment," said his wife, who met Altman in college. "But the worst part for him is his speech. He is such a social, outgoing person."

He has faced discrimination along the way, especially after leaving a Manhattan engineering job because he couldn't climb the subway stairs.

"He has such a hard time getting a job -- it was devastating," said Lorrie Altman. "On paper, he looked so good, but his speech was terrible. He has a college degree and isn't stupid, but all people see is the wheelchair."

Doctors say that many with the milder adult form of Tay-Sachs can lead full lives, despite their disability. And science is getting closer to finding treatments for this devastating disease.

Dr. Edwin Kolodny, former department chair and now professor of neurology at New York University School of Medicine, has been a leader in the field for 30 years. He first helped identify the role of the enzyme Hex-A and later tested more than 30,000 young adults in the 1970s and 1980s.

Today, he and others are involved in the promising gene therapy studies involving first mice, then cats and now sheep. Injecting genes into the brains of Jacob lambs has doubled their life span.

Clinical trials on humans are set to begin as soon as researchers can raise another $700,000 -- in addition to a grant from the National Institutes of Health -- to manufacture the vectors required to insert the genes into the body.

"It seems like every parent in the world would like to be part of the trial," said Kolodny. "And there are reasons to think there will be success here, especially for children who have a slightly later onset and not the classic form Tay-Sachs."

In the past, infantile Tay-Sachs has seen most of the medical attention. "These children have zero quality of life," he said.

Those with mild mutations, like Altman, who have 5 to 10 percent of Hex A enzyme activity, "sometimes lead full lives," according to Kolodny. "Intellectually, most of their cognitive function is retained. We have patients who are lawyers and accountants."

Pre-conception testing is still the gold standard for fighting the disease. "If your parents don't have the same recessive genes, you are home free," he said.

Those identified as at risk for having a child with Tay-Sachs can decide to adopt or conceive through in vitro fertilization, where geneticists can test the embryos before implantation to ensure the child will be disease-free.

Doctors can also do prenatal genetic testing and if the fetus is affected, the decision is up to the parents whether or not they want to terminate the pregnancy. "Three out of four times, they are reassured they have a normal child," said Kolodny.

Doctors say such testing -- at a cost of around $100 -- should be done routinely for 18 autosomal recessive disorders, including the gene for cystic fibrosis, which occurs in one in 20 caucasians, said Kolodny. Even with advances in Tay-Sachs testing in the Jewish community, public education must continue.

"The problem is each generation forgets what happened in the prior generation -- the grandmothers die out, " said Kolodny. "We need to educate health care professionals. Each new group of students graduating from medical school isn't prepared to ask the right questions."

Susan Kahn, NTSAD's executive director, who is involved in fundraising for research, agrees that along with a fight for a cure, genetic testing is critical.

"When there is a genetic disease, it's not just about that person, there is a whole implication for the rest of the family and how they deal with it," she said.

Stewart Altman sits on the association's board of directors and is a tireless crusader for a cure.

"He's got some disabilities that make it difficult for him to do certain things, but of all the board members asking for money to support, he is probably the boldest in our group," said Kahn. "He does have a lot of limitations, but he is still very energetic and wants to do something important. Not everyone responds with the same attitude."

His wife Lorrie backed that up with a laugh. "He is persistent," she said. "He carries these little envelopes around and will ask anyone he meets for a donation. It's almost embarrassing. He's not afraid to ask."

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Home | Gene Tools, LLC

Thursday, August 4th, 2016

What Do We Make?

Gene Tools makes Morpholino antisense oligos. Morpholino oligos bind to complementary RNA and get in the way of processes; they can knock down gene expression, modify RNA splicing or inhibit miRNA activity and maturation. Morpholinos are the premier knockdown tools used in developmental biology labs, the best RNA-blocking reagents for cells in culture and, as Vivo-Morpholinos, the most specific delivery-enhanced oligos available for other animal models. We are the sole commercial manufacturer selling research quantities of Morpholinos world-wide.

Morpholino oligos are short chains of about 25 Morpholino subunits. Each subunit is comprised of a nucleic acid base, a morpholine ring and a non-ionic phosphorodiamidate intersubunit linkage. Morpholinos do not degrade their RNA targets, but instead act via an RNAse H-independent steric blocking mechanism. With their requirement for greater complementarity with their target RNAs, Morpholinos are free of the widespread off-target expression modulation typical of knockdowns which rely on RISC or RNase-H activity. They are completely stable in cells and do not induce immune responses.

With their high mRNA binding affinity and exquisite specificity, Morpholinos yield reliable and predictable results. Depending on the oligo sequence selected, they either can block translation initiation in the cytosol (by targeting the 5' UTR through the first 25 bases of coding sequence), can modify pre-mRNA splicing in the nucleus (by targeting splice junctions or splice regulatory sites) or can inhibit miRNA maturation and activity (by targeting pri-miRNA or mature miRNA), as well as more exotic applications such as ribozyme inhibition, modifying poly-A tailing, blocking RNA translocation sequences or translational frameshifting. Morpholinos have been shown effective in animals, protists, plants and bacteria.

We are continually developing novel cytosolic delivery systems like our 'Endo-Porter' for cultured cells and our Vivo-Morpholinos for both cultures and in vivo delivery. With established delivery technologies it's easy to deliver Morpholinos into cultures, embryos or animals -- making Morpholinos the best tools for genetic studies and drug target validation programs.

What Sets Us Apart?

Morpholino oligos have excellent antisense properties compared to other gene knockdown systems. Microinjection or electroporation of Morpholino oligos into the embryos of frogs, zebrafish, chicks, sea urchins and other organisms successfully and specifically shuts down the expression of targeted genes, making Morpholinos an indispensable tool of developmental biologists. Morpholinos have also proven their versatility and efficacy in cultures of primary or immortal cells when delivered by Endo-Porter, electroporation or Vivo-Morpholinos. Usually, Vivo-Morpholinos are used to bring the specificity and efficacy of Morpholino oligos to experiments requiring systemic delivery in adult animals. The list of over 7500 publications using Morpholinos is growing daily and is maintained on-line in a browseable database.

Besides providing the best knockdown and splice modification tools, we also provide the best customer support available in the gene silencing industry. Our customer support team includes three Ph.D.-level scientists with hands-on Morpholino experience who are available to: 1) discuss your experiment design, 2) design your oligos for you, and 3) help you troubleshoot your experiments, all at no additional cost.

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Biotinidase Deficiency – GeneReviews – NCBI Bookshelf

Thursday, August 4th, 2016

Summary Clinical characteristics.

If untreated, young children with profound biotinidase deficiency usually exhibit neurologic abnormalities including seizures, hypotonia, ataxia, developmental delay, vision problems, hearing loss, and cutaneous abnormalities (e.g., alopecia, skin rash, candidiasis). Older children and adolescents with profound biotinidase deficiency often exhibit motor limb weakness, spastic paresis, and decreased visual acuity. Once vision problems, hearing loss, and developmental delay occur, they are usually irreversible, even with biotin therapy. Individuals with partial biotinidase deficiency may have hypotonia, skin rash, and hair loss, particularly during times of stress.

The diagnosis of biotinidase deficiency is established in a proband whose newborn screening or biochemical findings indicate multiple carboxylase deficiency based on either detection of deficient biotinidase enzyme activity in serum/plasma OR identification of biallelic pathogenic variants in BTD on molecular genetic testing.

Treatment of manifestations: All symptomatic children with profound biotinidase deficiency improve when treated with 5-10 mg of oral biotin per day. All individuals with profound biotinidase deficiency, even those who have some residual enzymatic activity, should have lifelong treatment with biotin. Children with vision problems may benefit from vision aids; those with hearing loss will usually benefit from hearing aids or cochlear implants, and those with developmental deficits from appropriate interventions.

Prevention of primary manifestations: Children with biotinidase deficiency identified by newborn screening should remain asymptomatic if biotin therapy is instituted early and continuously lifelong.

Surveillance: Annual vision and hearing evaluation, physical examination, and periodic assessment by a metabolic specialist.

Agents/circumstances to avoid: Raw eggs because they contain avidin, an egg-white protein that binds biotin and decreases the bioavailability of the vitamin.

Evaluation of relatives at risk: Testing of asymptomatic sibs of a proband ensures that biotin therapy for affected sibs can be instituted in a timely manner.

Biotinidase deficiency is inherited in an autosomal recessive manner. With each pregnancy, a couple who has had one affected child has a 25% chance of having an affected child, a 50% chance of having a child who is an asymptomatic carrier, and a 25% chance of having an unaffected child who is not a carrier. Carrier testing for at-risk family members and prenatal testing for pregnancies at increased risk are options if the pathogenic variants in the family are known.

Clinical issues and frequently asked questions regarding biotinidase deficiency have been addressed in a review [Wolf 2010].

Biotinidase deficiency should be suspected in infants with positive newborn screening results, untreated individuals with clinical findings, and persons with suggestive preliminary laboratory findings [Wolf 2012]:

Virtually 100% of infants with either profound biotinidase deficiency or partial biotinidase deficiency can be detected in the US by newborn screening (see National Newborn Screening Status Report).

Newborn screening utilizes a small amount of blood obtained from a heel prick for a colorimetric test for biotinidase activity:

Children or adults with untreated profound biotinidase deficiency usually exhibit one or more of the following non-specific features (which are also observed in many other inherited metabolic disorders):

Seizures

Hypotonia

Respiratory problems including hyperventilation, laryngeal stridor, and apnea

Developmental delay

Hearing loss

Vision problems, such as optic atrophy

Features more specific to profound biotinidase deficiency include the following:

Eczematous skin rash

Alopecia

Conjunctivitis

Candidiasis

Ataxia

Older children and adolescents may exhibit limb weakness, paresis, and scotomata. Some have exhibited findings suggestive of a myelopathy and have been initially incorrectly diagnosed and treated as having another disorder before biotinidase deficiency is correctly diagnosed [Wolf 2015].

Children or adults with untreated partial biotinidase deficiency may exhibit any of the above signs and symptoms, but the manifestations are mild and occur only when the person is stressed, such as with a prolonged infection.

The following findings are sugggestive of biotinidase deficiency:

Metabolic ketolactic acidosis

Organic aciduria (usually with the metabolites commonly seen in multiple carboxylase deficiency; however, 3-hydroxyisovalerate may be the only metabolite present). Note: Urinary organic acids can be normal even in individuals with biotinidase deficiency who are symptomatic.

Hyperammonemia

The diagnosis of biotinidase deficiency is established in a proband whose newborn screening or biochemical findings indicate multiple carboxylase deficiency based on either:

Biotinidase enzyme activity in serum. The working group of the American College of Medical Genetics Laboratory Quality Assurance Committee has established technical standards and guidelines for the diagnosis of biotinidase deficiency [Cowan et al 2010] (full text).

Molecular genetic testing is performed by single-gene testing. Sequence analysis of BTD is performed first, followed by gene-targeted deletion/duplication analysis if only one or no pathogenic variant is found.

Molecular Genetic Testing Used in Biotinidase Deficiency

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Test characteristics. See Clinical Utility Gene Card [Kry et al 2012] for information on test characteristics including sensitivity and specificity.

Individuals with biotinidase deficiency who are diagnosed before they have developed symptoms (e.g., by newborn screening) and who are treated with biotin have normal development [Mslinger et al 2001, Weber et al 2004] (see also Management, Prevention of Primary Manifestations). Neurologic problems occur only in those individuals with biotinidase deficiency who have recurrent symptoms and metabolic compromise prior to biotin treatment.

Early onset. Symptoms of untreated profound biotinidase deficiency (<10% mean normal serum biotinidase activity) usually appear between ages one week and ten years, with a mean age of three and one-half months [Wolf et al 1985b].

Some children with biotinidase deficiency manifest only a single finding, whereas others exhibit multiple neurologic and cutaneous findings.

The most common neurologic features in individuals with untreated, profound biotinidase deficiency are seizures and hypotonia [Wolf et al 1983a, Wolf et al 1985b, Wastell et al 1988, Wolf 1995, Wolf 2011]. The seizures are usually myoclonic but may be grand mal and focal; some children have infantile spasms [Salbert et al 1993b]. Some untreated children have exhibited spinal cord involvement characterized by progressive spastic paresis and myelopathy [Chedrawi et al 2008]. Older affected children often have ataxia and developmental delay.

Many symptomatic children with biotinidase deficiency exhibit a variety of central nervous system abnormalities on brain MRI or CT [Wolf et al 1983b, Wastell et al 1988, Lott et al 1993, Salbert et al 1993b, Grnewald et al 2004]. These findings may improve or become normal after biotin treatment.

Sensorineural hearing loss and eye problems (e.g., optic atrophy) have also been described in untreated children [Wolf et al 1983b, Taitz et al 1985, Salbert et al 1993a, Weber et al 2004]. Approximately 76% of untreated symptomatic children with profound biotinidase deficiency have sensorineural hearing loss that usually does not resolve or improve but remains static with biotin treatment [Wolf et al 2002].

Cutaneous manifestations include skin rash, alopecia, and recurrent viral or fungal infections caused by immunologic dysfunction.

Respiratory problems including hyperventilation, laryngeal stridor, and apnea can occur.

One death initially thought to be caused by sudden infant death syndrome was subsequently attributed to biotinidase deficiency [Burton et al 1987].

Late onset. A number of children with profound biotinidase deficiency were asymptomatic until adolescence, when they developed sudden loss of vision with progressive optic neuropathy and spastic paraparesis [Ramaekers et al 1992, Lott et al 1993, Ramaekers et al 1993]. After several months of biotin therapy, the eye findings resolved and the spastic paraparesis improved. In other individuals with enzyme deficiency, paresis and eye problems have occurred during early adolescence [Tokatli et al 1997, Wolf et al 1998, Wolf 2015].

Individuals with partial biotinidase deficiency (10%-30% of mean normal serum biotinidase activity) may develop symptoms only when stressed, such as during infection.

One child with partial biotinidase deficiency who was not treated with biotin exhibited hypotonia, skin rash, and hair loss during an episode of gastroenteritis at approximately age six months. When treated with biotin, the symptoms resolved.

Genotype/phenotype correlations are not well established. Deletions, insertions, or nonsense variants usually result in complete absence of biotinidase enzyme activity, whereas missense variants may or may not result in complete loss of biotinidase enzyme activity. Those with absence of all biotinidase enzyme activity are likely to be at increased risk for earlier onset of symptoms.

Although genotype-phenotype correlations are not well established, in one study, children with symptoms of profound biotinidase deficiency with null variants were more likely to develop hearing loss than those with missense variants, even if not treated for a period of time [Sivri et al 2007].

Certain genotypes correlate with complete biotinidase deficiency and others with partial biotinidase deficiency:

Profound biotinidase deficiency (<10% mean normal serum biotinidase activity):

Most BTD pathogenic variants cause complete loss or near-complete loss of biotinidase enzyme activity. These alleles are considered profound biotinidase deficiency alleles; a combination of two such alleles, whether homozygous or compound heterozygous, results in profound biotinidase deficiency. Affected individuals are likely to develop symptoms if not treated with biotin.

Partial biotinidase deficiency (10%-30% of mean normal serum biotinidase activity)

Heterozygotes

Individuals with one profound or one partial biotinidase deficiency BTD variant are carriers of biotinidase deficiency and do not exhibit symptoms [B Wolf, personal observation]. Such individuals do not require biotin therapy.

Individuals who are homozygous for the p.Asp444His pathogenic variant are expected to have approximately 45%-50% of mean normal serum biotinidase enzyme activity (which is similar to the activity of heterozygotes for profound biotinidase deficiency) and do not require biotin therapy.

Almost all children with profound biotinidase deficiency become symptomatic or are at risk of becoming symptomatic if not treated.

Several reports describe adults with profound biotinidase deficiency who have offspring who also have profound biotinidase deficiency identified by newborn screening, but who have never had symptoms [Wolf et al 1997, Baykal et al 2005]. In addition, several enzyme-deficient sibs of symptomatic children have apparently never exhibited symptoms. It is possible that these individuals would become symptomatic if stressed, such as with a prolonged infection.

Profound and partial biotinidase deficiency is the accepted nomenclature for this disorder.

Individuals with partial biotinidase deficiency were previously described as having late-onset or juvenile multiple or combined carboxylase deficiency.

Biotinidase deficiency should not be confused with holocarboxylase synthetase deficiency (see Differential Diagnosis), previously refered to as early-onset or infantile multiple or combined carboxylase deficiency.

Based on the results of worldwide screening of biotinidase deficiency [Wolf 1991], the incidence of the disorder is:

One in 137,401 for profound biotinidase deficiency;

One in 109,921 for partial biotinidase deficiency;

One in 61,067 for the combined incidence of profound and partial biotinidase deficiency.

The incidence of biotinidase deficiency is generally higher in populations with a high rate of consanguinity (e.g., Turkey, Saudi Arabia).

The incidence appears to be increased in the Hispanic population [Cowan et al 2012] and it may be lower in the African American population.

Carrier frequency in the general population is approximately one in 120.

Clinical features including vomiting, hypotonia, and seizures accompanied by metabolic ketolactic acidosis or mild hyperammonemia are often observed in inherited metabolic diseases. Individuals with biotinidase deficiency may exhibit clinical features that are misdiagnosed as other disorders (e.g., isolated carboxylase deficiency) before they are correctly identified [Suormala et al 1985, Wolf & Heard 1989]. Other symptoms that are more characteristic of biotinidase deficiency (e.g., skin rash, alopecia) can also occur in children with nutritional biotin deficiency, holocarboxylase synthetase deficiency, zinc deficiency, or essential fatty acid deficiency. See .

The biotin cycle

Free biotin enters the cycle from dietary sources or from the cleavage of biocytin or biotinyl-peptides by the action of biotinidase. The free biotin is then covalently attached to the various apocarboxylases, propionyl-CoA (more...)

Biotin deficiency. Biotin deficiency can usually be diagnosed by dietary history. Individuals with biotin deficiency may have a diet containing raw eggs or protracted parenteral hyperalimentation without biotin supplementation.

Low-serum biotin concentrations are useful in differentiating biotin and biotinidase deficiencies from holocarboxylase synthetase deficiency; however, it is important to know the method used for determining the biotin concentration as only methods that distinguish biotin from biocytin or bound biotin yield reliable estimates of free biotin concentrations.

Isolated carboxylase deficiency. Urinary organic acid analysis is useful for differentiating isolated carboxylase deficiencies from the multiple carboxylase deficiencies that occur in biotinidase deficiency and holocarboxylase synthetase deficiency:

The multiple carboxylase deficiencies are biotin responsive, whereas the isolated carboxylase deficiencies are not. A trial of biotin can be useful for discriminating between the disorders.

Isolated carboxylase deficiency can be diagnosed by demonstrating deficient enzyme activity of one of the three mitochondrial carboxylases in peripheral blood leukocytes (prior to biotin therapy) or in cultured fibroblasts grown in low biotin-containing medium, and normal activity of the other two carboxylases.

Holocarboxylase synthetase deficiency (OMIM). Both biotinidase deficiency and holocarboxylase synthetase deficiency are characterized by deficient activities of the three mitochondrial carboxylases in peripheral blood leukocytes prior to biotin treatment. In both disorders, these activities increase to near-normal or normal after biotin treatment.

The symptoms of biotinidase deficiency and holocarboxylase synthetase deficiency are similar, and clinical differentiation is often difficult.

The age of onset of symptoms may be useful for distinguishing between holocarboxylase synthetase deficiency and biotinidase deficiency. Holocarboxylase synthetase deficiency usually presents with symptoms before age three months, whereas biotinidase deficiency usually presents after age three months; however, there are exceptions for both disorders.

Organic acid abnormalities in biotinidase deficiency and holocarboxylase synthetase deficiency are similar and may be reported as consistent with multiple carboxylase deficiency. However, the tandem mass spectroscopic methodology that is being incorporated into many newborn screening programs should identify metabolites that are consistent with multiple carboxylase deficiency. Because most children with holocarboxylase synthetase deficiency excrete these metabolites in the newborn period, the disorder should be identifiable using this technology.

Definitive enzyme determinations are required to distinguish between the two disorders:

Individuals with holocarboxylase synthetase deficiency have deficient activities of the three mitochondrial carboxylases in extracts of fibroblasts that are incubated in medium containing only the biotin contributed by fetal calf serum (low biotin), whereas individuals with biotinidase deficiency have normal carboxylase activities in fibroblasts. The activities of the carboxylases in fibroblasts of individuals with holocarboxylase synthetase deficiency become near-normal to normal when cultured in medium supplemented with biotin (high biotin).

Sensorineural hearing loss (see Deafness and Hereditary Hearing Loss Overview). Sensorineural hearing loss has many causes. Biotinidase deficiency can be excluded as a cause by determining biotinidase enzyme activity in serum. This test should be performed specifically on children with hearing loss who are exhibiting other clinical features consistent with biotinidase deficiency.

To establish the extent of disease and needs in a symptomatic individual diagnosed with biotinidase deficiency, the following evaluations are recommended:

History of seizures, balance problems, feeding problems, breathing problems, loss of hair, fungal infections, skin rash, conjunctivitis

Physical examination for hypotonia, ataxia, eye findings such as optic atrophy, eczematous skin rash, alopecia, conjunctivitis, breathing abnormalities such as stridor, thrush, and/or candidiasis

Evaluation for psychomotor deficits

Evaluation for sensorineural hearing loss

Ophthalmologic examination

Identification of cellular immunologic abnormalities because of the increased risk of recurrent viral or fungal infections caused by immunologic dysfunction

Consultation with a metabolic specialist or clinical geneticist

To establish the extent of disease and needs in infants or children diagnosed with biotinidase deficiency following newborn screening, the following evaluations are recommended:

Physical examination for neurologic findings (e.g., hypotonia, ataxia), eye findings (e.g., conjunctivitis), skin findings (eczematous rash, alopecia), breathing abnormalities (e.g., stridor) and fungal infections caused by immunologic dysfunction (thrush and/or candidiasis).

Evaluation for psychomotor deficits

Evaluation for sensorineural hearing loss

Ophthalmologic examination (for finding such as optic atrophy)

Consultation with a metabolic specialist or clinical geneticist

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Biotinidase Deficiency - GeneReviews - NCBI Bookshelf

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Biology News Net – Latest Biology Articles, News & Current …

Thursday, August 4th, 2016

This is an illustration of SRM peaks and a human face. Reporting in the journal Cell, Senior Research Scientist Dr. Ulrike Kusebauch, of Institute for Systems Biology (ISB), describes the results of a collaboration between scientists at ISB, ETH Zurich and a number of other contributing institutes to develop the Human SRMAtlas, a compendium of proteomic assays for any human protein. The Human SRMAtlas is a compendium of highly specific mass spectrometry assays for the targeted identification and reproducible quantification of any protein in the predicted human proteome, including assays for many spliced variants, non-synonymous mutations and post-translational modifications. Using the technique called selected reaction monitoring, assays were developed with the use of 166,174 well-characterized, chemically synthesized proteotypic peptides. The SRMAtlas resource is freely publicly available at http://www.srmatlas.org and will equally benefit focused, hypothesis-driven and large proteome-scale studies. We expect this resource will significantly advance protein-based experimental biology to understand disease transitions and wellness trajectories because any human protein can now, in principle, be identified and quantified in any sample.

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Gene therapy – PBS

Saturday, October 24th, 2015

A treatment for Cystic Fibrosis. A cure for AIDS. The end of cancer. That's what the newspapers promised us in the early 1990's. Gene therapy was the answer to what ailed us. Scientists had at last learned how to insert healthy genes into unhealthy people. And those healthy genes would either replace the bad genes causing diseases like CF, sickle-cell anemia and hemophilia or stimulate the body's own immune system to rid itself of HIV and some forms of cancer. A decade later, none of these treatments have come to fruition and research into gene therapy has become politically unpopular, making clinical trials hard to approve and research dollars hard to come by. But some researchers who are taking a different approach to gene therapy could be on the road to more success than ever before. - - - - - - - - - - - -

Early Promise

Almost as soon as Watson and Crick unwound the double helix in the 1950's, researchers began considering the possibility- and ethics- of gene therapy. The goals were lofty- to fix inherited genetic diseases such as Cystic Fibrosis and hemophilia forever.

Gene therapists planned to isolate the relevant gene in question, prepare good copies of that gene, then deliver them to patients' cells. The hope was that the treated cells would give rise to new generations of healthy cells for the rest of the patient's life. The concept was elegant, but would require decades of research to locate the genes that cause illnesses.

By 1990, it was working in the lab. By inserting healthy genes into cells from CF patients, scientists were able to transmogrify the sick cells as if by magic into healthy cells.

That same year, four-year-old Ashanti DeSilva became the first person in history to receive gene therapy. Dr. W. French Anderson of the National Heart, Lung and Blood Institute and Dr. Michael Blaese and Dr. Kenneth Culver, both of the National Cancer Institute, performed the historic and controversial experiment.

DeSilva suffered from a rare immune disorder known as ADA deficiency that made her vulnerable to even the mildest infections. A single genetic defect- like a typo in a novel- left DeSilva unable to produce an important enzyme. Without that enzyme, DeSilva was likely to die a premature death.

Anderson, Blaese and Culver drew the girl's blood and treated her defective white blood cells with the gene she lacked. The altered cells were then injected back into the girl, where- the scientists hoped- they would produce the enzyme she needed as well as produce future generations of normal cells.

Though the treatment proved safe, its efficacy is still in question. The treated cells did produce the enzyme, but failed to give rise to healthy new cells. DeSilva, who is today relatively healthy, still receives periodic gene therapy to maintain the necessary levels of the enzyme in her blood. She also takes doses of the enzyme itself, in the form of a drug called PEG-ADA, which makes it difficult to tell how well the gene therapy would have worked alone.

"It was a very logical approach," says Dr. Jeffrey Isner, Chief of Vascular Medicine and Cardiovascular Research at St. Elizabeth's Medical Center in Boston as well as Professor of Medicine at Tufts University School of Medicine. "But in most cases the strategy failed, because the vectors we have today are not ready for prime time." - - - - - - - - - - - - 4 pages: | 1 | 2 | 3 | 4 |

Photo: Dr. W. French Anderson

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Gene therapy - PBS

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Gene therapy | Cancer Research UK

Thursday, October 22nd, 2015

Researchers are looking at different ways of using gene therapy, including

Some types of gene therapy aim to boost the body's natural ability to attack cancer cells. Our immune system has cells that recognise and kill harmful things that can cause disease, such as cancer cells.

There are many different types of immune cell. Some of them produce proteins that encourage other immune cells to destroy cancer cells. Some types of therapy add genes to a patient's immune cells to make them better at finding or destroying particular types of cancer. There are a few trials using this type of gene therapy in the UK.

Some gene therapies put genes into cancer cells to make the cells more sensitive to particular treatments such as chemotherapy or radiotherapy. This type of gene therapy aims to make the other cancer treatments work better.

Some types of gene therapy deliver genes into the cancer cells that allow the cells to change drugs from an inactive form to an active form. The inactive form of the drug is called a pro drug.

After giving the carrier containing the gene, the doctor gives the patient the pro drug. The pro drug may be a tablet or capsule that you swallow, or you may have it into the bloodstream.

The pro drug circulates in the body and doesn't harm normal cells. But when it reaches the cancer cells, the gene activates it and the drug kills the cancer cells.

Some gene therapies block processes that cancer cells use to survive. For example, most cells in the body are programmed to die if their DNA is damaged beyond repair. This is called programmed cell death or apoptosis. But cancer cells block this process so they don't die even when they are supposed to. Some gene therapy strategies aim to reverse this blockage. Doctors hope that these new types of treatment will make the cancer cells die.

Some viruses infect and kill cells. Researchers are working on ways to change these viruses so that they only target and kill cancer cells, leaving healthy cells alone. This sort of treatment uses the viruses to kill cancer cells directly rather than to deliver genes. So it is not cancer gene therapy in the true sense of the word. But doctors sometimes refer to it as gene therapy.

One example of this type of research uses the cold sore virus (herpes simplex virus). The changed virus is called Oncovex. It has been tested in early clinical trials for advanced melanoma, pancreatic cancer and head and neck cancers.

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Gene therapy – An Introduction to Genetic Analysis – NCBI …

Wednesday, October 14th, 2015

The general approach of gene therapy is nothing more than an extension of the technique for clone selection by functional complementation (Chapter 12). The functions absent in the recipient as a result of a defective gene are introduced on a vector that inserts into one of the recipients chromosomes and thereby generates a transgenic animal that has been genetically cured. The technique is of great potential in humans because it offers the hope of correcting hereditary diseases. However, gene therapy is also being applied to mammals other than humans.

The first example of gene therapy in a mammal was the correction of a growth-hormone deficiency in mice. The recessive mutation little (lit) results in dwarf mice. Even though a mouses growth-hormone gene is present and apparently normal, no mRNA for this gene is produced. The initial step in correcting this deficiency was to inject homozygous lit/lit eggs with about 5000 copies of a 5-kb linear DNA fragment that contained the rat growth-hormone structural gene (RGH) fused to a regulatorpromoter sequence from a mouse metallothionein gene (MP). The normal job of metallothionein is to detoxify heavy metals, so the regulatory sequence is responsive to the presence of heavy metals in the animal. The eggs were then implanted into pseudopregnant mice, and the baby mice were raised. About 1 percent of these babies turned out to be transgenic, showing increased size when heavy metals were administered in the course of development. A representative transgenic mouse was then crossed with a homozygous lit/lit female. The ensuing pedigree is shown in . We can see in that mice two to three times the weight of their lit/lit relatives are produced in subsequent generations, with the rat growth-hormone transgene acting as a dominant allele, always heterozygous in this pedigree. The rat growth-hormone transgene also makes lit+ mice bigger ().

The rat growth-hormone gene (RGH), under the control of a mouse promoter region that is responsive to heavy metals, is inserted into a plasmid and used to produce a transgenic mouse. RGH compensates for the inherent dwarfism (lit/lit) in the mouse. RGH (more...)

Transgenic mouse. The mice are siblings, but the mouse on the left was derived from an egg transformed by injection with a new gene composed of the mouse metallothionein promoter fused to the rat growth-hormone structural gene. (This mouse weighs 44g, (more...)

The site of insertion of the introduced DNA in mammals is highly variable, and the DNA is generally not found at the homologous locus. Hence, gene therapy most often provides not a genuine correction of the original problem but a masking of it.

Similar technology has been used to develop transgenic fast-growing strains of Pacific salmon, with spectacular results. A plasmid containing a growth-hormone gene placed next to a metallothionein promoter (all derived from salmon) was microinjected into salmon eggs. A small proportion of the resulting fish proved to be transgenic, testing positive when their DNA was probed with the plasmid construct. These fish were on average 11-fold heavier than the nontransgenic controls (). Progeny inherited the transgene in the same manner as the mice in the earlier example.

Effect of introducing a hormone transgene complex with a strong promoter into Pacific salmon. All salmon shown are the same age. (R. H. Devlin, T. Y. Yesaki, C. A. Biagi, E. M. Donaldson, P. Swanson, and W.-K. Chan, Extraordinary Growth, (more...)

Perhaps the most exciting and controversial application of transgenic technology is in human gene therapy, the treatment and alleviation of human genetic disease by adding exogenous wild-type genes to correct the defective function of mutations. We have seen that the first case of gene therapy in mammals was to cure a genotypically dwarf fertilized mouse egg by injecting the appropriate wild-type allele for normal growth. This technique () has little application in humans, because it is currently impossible to diagnose whether a fertilized egg cell carries a defective genotype without destroying the cell. (However, in an early embryo containing only a few cells, one cell can be removed and analyzed with no ill effects on the remainder.)

Two basic types of gene therapy can be applied to humans, germ line and somatic. The goal of germ-line gene therapy () is the more ambitious: to introduce transgenic cells into the germ line as well as into the somatic cell population. Not only should this therapy achieve a cure of the person treated, but some gametes could also carry the corrected genotype. We have seen that such germinal therapy has been achieved by injecting mice eggs. However, the protocol that is relevant for application to humans is the removal of an early embryo (blastocyst) with a defective genotype from a pregnant mouse and injection with transgenic cells containing the wild-type allele. These cells become part of many tissues of the body, often including the germ line, which will give rise to the gonads. Then the gene can be passed on to some or all progeny, depending on the size of the clone of transgenic cells that lodges in the germinal area. However, no human germ-line gene therapy has been performed to date.

We have seen that most transforming fragments will insert ectopically throughout the genome. This is a disadvantage in human gene therapy not only because of the possibility of the ectopic insert causing gene disruption, but also because, even if the disease phenotype is reversed, the defective allele is still present and can segregate away from the transgene in future generations. Therefore, for effective germinal gene therapy, an efficient targeted gene replacement will be necessary, in which case the wild-type transgene replaces the resident defective copy by a double crossover.

Somatic gene therapy () focuses only on the body (soma). The approach is to attempt to correct a disease phenotype by treating some somatic cells in the affected person. At present, it is not possible to render an entire body transgenic, so the method addresses diseases whose phenotype is caused by genes that are expressed predominantly in one tissue. In such cases, it is likely that not all the cells of that tissue need to become transgenic; a percentage of cells being transgenic can ameliorate the overall disease symptoms. The method proceeds by removing some cells from a patient with the defective genotype and making these cells transgenic through the introduction of copies of the cloned wild-type gene. The transgenic cells are then reintroduced into the patients body, where they provide normal gene function.

Currently, there are two ways of getting the transgene into the defective somatic cells. Both methods use viruses. The older method uses a disarmed retrovirus with the transgene spliced into its genome, replacing most of the viral genes. The natural cycle of retroviruses includes the integration of the viral genome at some location in one of the host cells chromosomes. The recombinant retrovirus will carry the transgene along with it into the chromosome. This type of vector poses a potential problem, because the integrating virus can act as an insertional mutagen and inactivate some unknown resident gene, causing a mutation. Another problem with this type of vector is that a retrovirus attacks only proliferating cells such as blood cells. This procedure has been used for somatic gene therapy of severe combined immunodeficiency disease (SCID), otherwise known as bubble-boy disease. This disease is caused by a mutation in the gene encoding the blood enzyme adenosine deaminase (ADA). In an attempt at gene therapy, blood stem cells are removed from the bone marrow, the transgene is added, and the transgenic cells are reintroduced into the blood system. Prognosis for such patients is currently good.

Even solid tissues seem to be accessible to somatic gene therapy. In a dramatic case, gene therapy was administered to a patient homozygous for a recessive mutant allele of the LDLR gene for low-density-lipoprotein receptor (genotype LDLR/LDLR). This mutant allele increases the risk of atherosclerosis and coronary disease. The receptor protein is made in liver cells, so 15 percent of the patients liver was removed, and the liver cells were dissociated and treated with retrovirus carrying the LDLR+ allele. Transgenic cells were reintroduced back into the body by injection into the portal venous system, which takes blood from the intestine to the liver. The transgenic cells took up residence in the liver. The latest reports are that the procedure seems to be working and the patients lipid profile has improved.

The other vector used in human gene therapy is the adenovirus. This virus normally attacks respiratory epithelia, injecting its genome into the epithelial cells. The viral genome does not integrate into a chromosome but persists extrachromosomally in the cells, which eliminates the problem of insertional mutagenesis by the vector. Another advantage of the adenovirus as a vector is that it attacks nondividing cells, making most tissues susceptible in principle. Inasmuch as cystic fibrosis is a disease of the respiratory epithelium, adenovirus is an appropriate choice of vector for treating this disease, and gene therapy for cystic fibrosis is currently being attempted with the use of this vector. Viruses bearing the wild-type cystic fibrosis allele are introduced through the nose as a spray. It is also possible to use the adenovirus to attack cells of the nervous system, muscle, and liver.

A promising type of construct that should find use in gene therapy is the human artificial chromosome (HAC). HACs contain essentially the same components as YACs. They were made by mixing human telomeric DNA, genomic DNA, and arrays of repetitive -satellite DNA (thought to have centrometric activity). To this unjoined mixture was added lipofectin, a substance needed for passage through the membrane, and the complete mixture was added to cultured cells. Some cells were observed to contain small new chromosomes that seemed to have assembled de novo inside the cell from the added components (). When the technology has been perfected, these HACs should be potent vectors capable of transferring large amounts of human DNA into cells in a stable replicating form.

An artificial human chromosome (at arrow). (John J. Harrington, G. Van Bokkelen, R. W. Mays, K. Gustashaw, and H. F. Willard, Formation of de Novo Centromeres and Construction of First-Generation Human Artificial Microchromosomes, Nature (more...)

Gene therapy introduces transgenic cells either into somatic tissue to correct defective function (somatic therapy) or into the germ line for transmission to descendants (germ-line therapy).

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Gene Therapy Successes – Learn Genetics

Monday, October 12th, 2015

Researchers have been working for decades to bring gene therapy to the clinic, yet very few patients have received any effective gene-therapy treatments. But that doesn't mean gene therapy is an impossible dream. Even though gene therapy has been slow to reach patients, its future is very encouraging. Decades of research have taught us a lot about designing safe and effective vectors, targeting different types of cells, and managing and minimizing immune responses in patients. We've also learned a lot about the disease genes themselves. Today, many clinical trials are underway, where researchers are carefully testing treatments to ensure that any gene therapy brought into the clinic is both safe and effective.

Below are some gene therapy success stories. Successes represent a variety of approachesdifferent vectors, different target cell populations, and both in vivo and ex vivo approachesto treating a variety of disorders.

Sebastian Misztal was a patient in a hemophilia gene therapy trial in 2011. Following the treatment, Misztal no longer had spontaneous bleeding episodes. Credit: UCLH/UCL NIHR Biomedical Research Centre

Several inherited immune deficiencies have been treated successfully with gene therapy. Most commonly, blood stem cells are removed from patients, and retroviruses are used to deliver working copies of the defective genes. After the genes have been delivered, the stem cells are returned to the patient. Because the cells are treated outside the patient's body, the virus will infect and transfer the gene to only the desired target cells.

Severe Combined Immune Deficiency (SCID) was one of the first genetic disorders to be treated successfully with gene therapy, proving that the approach could work. However, the first clinical trials ended when the viral vector triggered leukemia (a type of blood cancer) in some patients. Since then, researchers have begun trials with new, safer viral vectors that are much less likely to cause cancer.

Adenosine deaminase (ADA) deficiency is another inherited immune disorder that has been successfully treated with gene therapy. In multiple small trials, patients' blood stem cells were removed, treated with a retroviral vector to deliver a functional copy of the ADA gene, and then returned to the patients. For the majority of patients in these trials, immune function improved to the point that they no longer needed injections of ADA enzyme. Importantly, none of them developed leukemia.

Gene therapies are being developed to treat several different types of inherited blindnessespecially degenerative forms, where patients gradually lose the light-sensing cells in their eyes. Encouraging results from animal models (especially mouse, rat, and dog) show that gene therapy has the potential to slow or even reverse vision loss.

The eye turns out to be a convenient compartment for gene therapy. The retina, on the inside of the eye, is both easy to access and partially protected from the immune system. And viruses can't move from the eye to other places in the body. Most gene-therapy vectors used in the eye are based on AAV (adeno-associated virus).

In one small trial of patients with a form of degenerative blindness called LCA (Leber congenital amaurosis), gene therapy greatly improved vision for at least a few years. However, the treatment did not stop the retina from continuing to degenerate. In another trial, 6 out of 9 patients with the degenerative disease choroideremia had improved vision after a virus was used to deliver a functional REP1 gene.

Credit: Jean Bennett, MD, PhD, Perelman School of Medicine, University of Pennsylvania; Manzar Ashtari, Ph.D., of The Children's Hospital of Philadelphia, Science Translational Medicine.

People with hemophilia are missing proteins that help their blood form clots. Those with the most-severe forms of the disease can lose large amounts of blood through internal bleeding or even a minor cut.

In a small trial, researchers successfully used an adeno-associated viral vector to deliver a gene for Factor IX, the missing clotting protein, to liver cells. After treatment, most of the patients made at least some Factor IX, and they had fewer bleeding incidents.

Patients with beta-Thalassemia have a defect in the beta-globin gene, which codes for an oxygen-carrying protein in red blood cells. Because of the defective gene, patients don't have enough red blood cells to carry oxygen to all the body's tissues. Many who have this disorder depend on blood transfusions for survival.

In 2007, a patient received gene therapy for severe beta-Thalassemia. Blood stem cells were taken from his bone marrow and treated with a retrovirus to transfer a working copy of the beta-globin gene. The modified stem cells were returned to his body, where they gave rise to healthy red blood cells. Seven years after the procedure, he was still doing well without blood transfusions.

A similar approach could be used to treat patients with sickle cell disease.

In 2012, Glybera became the first viral gene-therapy treatment to be approved in Europe. The treatment uses an adeno-associated virus to deliver a working copy of the LPL (lipoprotein lipase) gene to muscle cells. The LPL gene codes for a protein that helps break down fats in the blood, preventing fat concentrations from rising to toxic levels.

Several promising gene-therapy treatments are under development for cancer. One, a modified version of the herpes simplex 1 virus (which normally causes cold sores) has been shown to be effective against melanoma (a skin cancer) that has spread throughout the body. The treatment, called T-VEC, uses a virus that has been modified so that it will (1) not cause cold sores; (2) kill only cancer cells, not healthy ones; and (3) make signals that attract the patient's own immune cells, helping them learn to recognize and fight cancer cells throughout the body. The virus is injected directly into the patient's tumors. It replicates (makes more of itself) inside the cancer cells until they burst, releasing more viruses that can infect additional cancer cells.

A completely different approach was used in a trial to treat 59 patients with leukemia, a type of blood cancer. The patients' own immune cells were removed and treated with a virus that genetically altered them to recognize a protein that sits on the surface of the cancer cells. After the immune cells were returned to the patients, 26 experienced complete remission.

Patients with Parkinson's disease gradually lose cells in the brain that produce the signaling molecule dopamine. As the disease advances, patients lose the ability to control their movements.

A small group of patients with advanced Parkinson's disease were treated with a retroviral vector to introduce three genes into cells in a small area of the brain. These genes gave cells that don't normally make dopamine the ability to do so. After treatment, all of the patients in the trial had improved muscle control.

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Types of Gene Therapy Treatment | MD Anderson Cancer Center

Tuesday, September 29th, 2015

Much of today's cancer research is devoted to finding missing or defective genes that cause cancer or increase an individual's risk for certain types of cancer. Gene research at MDAnderson has resulted in many important discoveries. We identified the mutated multiple advanced cancers gene (MMAC1) involved in some common cancers. We also performed the first successful correction of a defective tumor suppressor gene (p53) in human lung cancer. Current gene therapies are experimental, and many are still tested only on animals. There are some clinical trials involving a very small number of human subjects.

The potential benefits of gene therapy are two-fold:

The focus of most gene therapy research is the replacement of a missing or defective gene with a functional, healthy copy, which is delivered to target cells with a "vector." Viruses are commonly used as vectors because of their ability to penetrate a cells DNA. These vector viruses are inactivated so they cannot reproduce and cause disease. Gene transfer therapy can be done outside the body (ex vivo) by extracting bone marrow or blood from the patient and growing the cells in a laboratory. The corrected copy of the gene is introduced and allowed to penetrate the cells DNA before being injected back into the body. Gene transfers can also be done directly inside the patients body (in vivo).

Other therapies include:

Gene therapy is a complicated area of research, and many questions remain unanswered. Some cancers are caused by more than one gene, and some vectors, if used incorrectly, can actually cause cancer or other diseases. Replacing faulty genes with working copies also brings up ethical issues that must be addressed before these therapies can be accepted for preventing cancer. Talk to your cancer specialist about the implications of gene therapy.

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Gene Therapy – A Revolution in Progress: Human Genetics …

Sunday, September 27th, 2015

Gene therapy attempts to treat genetic diseases at the molecular level by correcting what is wrong with defective genes. Clinical research into gene therapys safety and effectiveness has just begun. No one knows if gene therapy will work, or for what diseases. If gene therapy is successful, it could work by preventing a protein from doing something that causes harm, restoring the normal function of a protein, giving proteins new functions, or enhancing the existing functions of proteins. How Do You Do It? Gene therapy relies on finding a dependable delivery system to carry the correct gene to the affected cells. The gene must be delivered inside the target cells and work properly without causing adverse effects. Delivering genes that will work correctly for the long term is the greatest challenge of gene therapy.

Human ex vivo Gene Therapy

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How does gene therapy work? – Genetics Home Reference

Wednesday, September 2nd, 2015

Gene therapy is designed to introduce genetic material into cells to compensate for abnormal genes or to make a beneficial protein. If a mutated gene causes a necessary protein to be faulty or missing, gene therapy may be able to introduce a normal copy of the gene to restore the function of the protein.

A gene that is inserted directly into a cell usually does not function. Instead, a carrier called a vector is genetically engineered to deliver the gene. Certain viruses are often used as vectors because they can deliver the new gene by infecting the cell. The viruses are modified so they cant cause disease when used in people. Some types of virus, such as retroviruses, integrate their genetic material (including the new gene) into a chromosome in the human cell. Other viruses, such as adenoviruses, introduce their DNA into the nucleus of the cell, but the DNA is not integrated into a chromosome.

The vector can be injected or given intravenously (by IV) directly into a specific tissue in the body, where it is taken up by individual cells. Alternately, a sample of the patients cells can be removed and exposed to the vector in a laboratory setting. The cells containing the vector are then returned to the patient. If the treatment is successful, the new gene delivered by the vector will make a functioning protein.

Researchers must overcome many technical challenges before gene therapy will be a practical approach to treating disease. For example, scientists must find better ways to deliver genes and target them to particular cells. They must also ensure that new genes are precisely controlled by the body.

A new gene is injected into an adenovirus vector, which is used to introduce the modified DNA into a human cell. If the treatment is successful, the new gene will make a functional protein.

The Genetic Science Learning Center at the University of Utah provides information about various technical aspects of gene therapy in Gene Delivery: Tools of the Trade. They also discuss other approaches to gene therapy and offer a related learning activity called Space Doctor.

The Better Health Channel from the State Government of Victoria (Australia) provides a brief introduction to gene therapy, including the gene therapy process and delivery techniques.

Penn Medicines Oncolink describes how gene therapy works and how it is administered to patients.

Next: Is gene therapy safe?

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Recent Articles | Gene Therapy | The Scientist Magazine

Sunday, August 23rd, 2015

Most Recent

By targeting rhodopsin genes to neurons, scientists help blind mice see.

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Researchers deploy ancestors of todays adeno-associated viruses to deliver gene therapies without immune system interference.

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Expressing a gene for a component of the inner ears hair cells treated a form of genetic deafness.

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The results of a Phase 2 trial suggest that delivering normal copies of the gene that causes cystic fibrosis may slow lung decline.

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By Kerry Grens | June 26, 2015

Biotech firm likely to pull the plug after its gene therapy product fails.

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Participants of two gene-therapy trials who experienced partial restoration of sight following treatment are now losing their vision once again.

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A newly discovered protein promotes immunity to viruses and cancer by triggering the production of cytotoxic T cells.

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By Kerry Grens | January 22, 2015

In a mouse model of a rare disease, scientists have figured out how to reduce the elevated cancer risk tied to a gene therapy treatment.

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A small peptide helps a silencing construct home in on the adipocytes of obese mice.

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A former postdoc in a prominent gene therapy lab is branded a fraud by the US government more than three years after having a slew of papers retracted from various journals.

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Gene Therapy I – RCN

Saturday, August 8th, 2015

Many human diseases are caused by defective genes.

All of these diseases are caused by a defect at a single gene locus. (The inheritance is recessive so both the maternal and paternal copies of the gene must be defective.) Is there any hope of introducing functioning genes into these patients to correct their disorder? Probably.

Other diseases also have a genetic basis, but it appears that several genes must act in concert to produce the disease phenotype. The prospects of gene therapy in these cases seems far more remote.

It is a disease of young children because, until recently, the absence of an immune system left them prey to infections that ultimately killed them.

Once the virus has infected the target cells, this RNA is reverse transcribed into DNA and inserted into the chromosomal DNA of the host.

The first attempts at gene therapy for SCID children (in 1990), used their own T cells (produced following ADA-PEG therapy) as the target cells.

In June of 2002, a team of Italian and Israeli doctors reported on two young SCID patients that were treated with their own blood stem cells that had been transformed in vitro with a retroviral vector carrying the ADA gene. After a year, both children had fully-functioning immune systems (T, B, and NK cells) and were able to live normal lives without any need for treatment with ADA-PEG or immune globulin (IG). The doctors attribute their success to first destroying some of the bone marrow cells of their patients to "make room" for the transformed cells.

Nine years later (August 2011) these two patients are still thriving and have been joined by 28 other successfully-treated children most of whom no longer need to take ADA-PEG.

Gene therapy has also succeeded for 20 baby boys who suffered from another form of severe combined immunodeficiency called X-linked SCID because it is caused by a mutated X-linked gene encoding a subunit called c (gamma-c) of the receptor for several interleukins, including interleukin-7 (IL-7).

IL-7 is essential for converting blood stem cells into the progenitors of T cells. [View]. Boys with X-linked SCID can make normal B cells, but because B cells need T-helper cells to function, these boys could make neither cell-mediated nor antibody-mediated immune responses and had to live in a sterile bubble before their treatment.

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Gene Therapy and Children – KidsHealth

Sunday, July 12th, 2015

Gene therapy carries the promise of cures for many diseases and for types of medical treatment that didn't seem possible until recently. With its potential to eliminate and prevent hereditary diseases such as cystic fibrosis and hemophilia and its use as a possible cure for heart disease, AIDS, and cancer, gene therapy is a potential medical miracle-worker.

But what about gene therapy for children? There's a fair amount of risk involved, so thus far only seriously ill kids or those with illnesses that can't be cured by standard medical treatments have been involved in clinical trials using gene therapy.

As those studies continue, gene therapy may soon offer hope for children with serious illnesses that don't respond to conventional therapies.

Our genes help make us unique. Inherited from our parents, they go far in determining our physical traits like eye color and the color and texture of our hair. They also determine things like whether babies will be male or female, the amount of oxygen blood can carry, and the likelihood of getting certain diseases.

Genes are composed of strands of a molecule called DNA and are located in single file within the chromosomes. The genetic message is encoded by the building blocks of the DNA, which are called nucleotides. Approximately 3 billion pairs of nucleotides are in the chromosomes of a human cell, and each person's genetic makeup has a unique sequence of nucleotides. This is mainly what makes us different from one another.

Scientists believe that every human has about 25,000 genes per cell. A mutation, or change, in any one of these genes can result in a disease, physical disability, or shortened life span. These mutations can be passed from one generation to another, inherited just like a mother's curly hair or a father's brown eyes. Mutations also can occur spontaneously in some cases, without having been passed on by a parent. With gene therapy, the treatment or elimination of inherited diseases or physical conditions due to these mutations could become a reality.

Gene therapy involves the manipulation of genes to fight or prevent diseases. Put simply, it introduces a "good" gene into a person who has a disease caused by a "bad" gene.

The two forms of gene therapy are:

Currently, gene therapy is done only through clinical trials, which often take years to complete. After new drugs or procedures are tested in laboratories, clinical trials are conducted with human patients under strictly controlled circumstances. Such trials usually last 2 to 4 years and go through several phases of research. In the United States, the U.S. Food and Drug Administration (FDA) must then approve the new therapy for the marketplace, which can take another 2 years.

The most active research being done in gene therapy for kids has been for genetic disorders (like cystic fibrosis). Other gene therapy trials involve children with severe immunodeficiencies, such as adenosine deaminase (ADA) deficiency (a rare genetic disease that makes kids prone to serious infection), sickle cell anemia, thalassemia, hemophilia, and those with familial hypercholesterolemia (extremely high levels of serum cholesterol).

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Challenges in Gene Therapy – Learn Genetics

Friday, July 3rd, 2015

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Gene Therapy

Challenges in Gene Therapy?

Gene therapy is not a new field; it has been evolving for decades. Despite the best efforts of researchers around the world, however, gene therapy has seen only limited success. Why?

Gene therapy poses one of the greatest technical challenges in modern medicine. It is very hard to introduce new genes into cells of the body and keep them working. And there are financial concerns: Can a company profit from developing a gene therapy to treat a rare disorder? If not, who will develop and pay for these life-saving treatments?

Let's look at some of the main challenges in gene therapy.

For some disorders, gene therapy will work only if we can deliver a normal gene to a large number of cellssay several millionin a tissue. And they have to the correct cells, in the correct tissue. Once the gene reaches its destination, it must be activated, or turned on, to make the protein it encodes. And once it's turned on, it must remain on; cells have a habit of shutting down genes that are too active or exhibiting other unusual behaviors.

Introducing changes into the wrong cells Targeting a gene to the correct cells is crucial to the success of any gene therapy treatment. Just as important, though, is making sure that the gene is not incorporated into the wrong cells. Delivering a gene to the wrong tissue would be inefficient, and it could cause health problems for the patient.

For example, improper targeting could incorporate the therapeutic gene into a patient's germline, or reproductive cells, which ultimately produce sperm and eggs. Should this happen, the patient would pass the introduced gene to his or her children. The consequences would vary, depending on the gene.

Our immune systems are very good at fighting off intruders such as bacteria and viruses. Gene-delivery vectors must be able to avoid the body's natural surveillance system. An unwelcome immune response could cause serious illness or even death.

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Gene Therapy – Nature

Thursday, July 2nd, 2015

Included in top indexing databases and has an impact factor of 3.104! The journal website is hosted on http://www.nature.com that gets 8.4 million page views per month Over 69,000 table of contents registrants and 82,706 page views across the Gene Therapy web site on average Easy and quick online submission system Rapid and rigorous peer review Advanced Article Preview (AAP) feature to be available from early March 2015 to all accepted original and review article authors. Accepted articles to be available online within 72 hours of acceptance into production! For further information on AAP please refer to About accepted article preview section Free manuscript deposition service to Pub Med Central on behalf of non-open access authors We regularly promote content alongside that of the Nature branded titles on our subject pages and in collections as well as via social media We also have a number of regional websites reaching a wide and varied audience demographic http://www.nature.com/regions Free online issue

Volume 22, No 7 July 2015 ISSN: 0969-7128 EISSN: 1476-5462

2014 Impact Factor 3.104* 117/289 Biochemistry & Molecular Biology 47/162 Biotechnology & Applied Microbiology 63/167 Genetics & Heredity 43/123 Medicine, Research & Experimental

Editors: J Glorioso, USA N Lemoine, UK

*2014 Journal Citation Reports Science Edition (Thomson Reuters, 2015)

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Gene Therapy now offers authors the option to publish their articles with immediate open access upon publication. Open access articles will also be deposited on PubMed Central at the time of publication and will be freely available immediately. Find out more from our FAQs page.

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Gene therapy – Science Daily

Friday, May 22nd, 2015

Gene therapy is the insertion of genes into an individual's cells and tissues to treat a disease, and hereditary diseases in which a defective mutant allele is replaced with a functional one.

Although the technology is still in its infancy, it has been used with some success.

Antisense therapy is not strictly a form of gene therapy, but is a genetically-mediated therapy and is often considered together with other methods.

In most gene therapy studies, a "normal" gene is inserted into the genome to replace an "abnormal," disease-causing gene.

A carrier called a vector must be used to deliver the therapeutic gene to the patient's target cells.

Currently, the most common type of vectors are viruses that have been genetically altered to carry normal human DNA.

Viruses have evolved a way of encapsulating and delivering their genes to human cells in a pathogenic manner.

Scientists have tried to harness this ability by manipulating the viral genome to remove disease-causing genes and insert therapeutic ones.

Target cells such as the patient's liver or lung cells are infected with the vector.

The vector then unloads its genetic material containing the therapeutic human gene into the target cell.

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What is gene therapy? – Genetics Home Reference

Tuesday, May 19th, 2015

Gene therapy is an experimental technique that uses genes to treat or prevent disease. In the future, this technique may allow doctors to treat a disorder by inserting a gene into a patients cells instead of using drugs or surgery. Researchers are testing several approaches to gene therapy, including:

Replacing a mutated gene that causes disease with a healthy copy of the gene.

Inactivating, or knocking out, a mutated gene that is functioning improperly.

Introducing a new gene into the body to help fight a disease.

Although gene therapy is a promising treatment option for a number of diseases (including inherited disorders, some types of cancer, and certain viral infections), the technique remains risky and is still under study to make sure that it will be safe and effective. Gene therapy is currently only being tested for the treatment of diseases that have no other cures.

MedlinePlus from the National Library of Medicine offers a list of links to information about genes and gene therapy.

Educational resources related to gene therapy are available from GeneEd.

The Genetic Science Learning Center at the University of Utah provides an interactive introduction to gene therapy and a discussion of several diseases for which gene therapy has been successful.

The Centre for Genetics Education provides an introduction to gene therapy, including a discussion of ethical and safety considerations.

KidsHealth from Nemours offers a fact sheet called Gene Therapy and Children.

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What is Gene Therapy ? – Learn Genetics

Tuesday, May 19th, 2015

Gene therapy could be a way to fix a genetic problem at its source. By adding a corrected copy of a defective gene, gene therapy promises to help diseased tissues and organs work properly. This approach is different from traditional drug-based approaches, which may treat symptoms but not the underlying genetic problems.

Most commonly, gene therapy uses a vector, typically a virus, to deliver a gene to the cells where it's needed. Once it's inside, the cell's gene-reading machinery uses the information in the gene to build RNA and protein molecules. The proteins (or RNA) can then carry out their job in the cells.

But gene therapy is not a molecular bandage that will automatically fix any genetic problem. While many disorders or medical conditions can potentially be treated using gene therapy, others are not suitable for this approach. So what makes a condition a good candidate for gene therapy?

Could the condition be corrected by adding one or a few functional genes? For you to even consider gene therapy, the answer must be "yes." For instance, genetic disorders caused by mutations in single genes tend to be good candidates for gene therapy, while diseases involving many genes and environmental factors tend to be poor candidates.

Do you know which genes are involved? If you plan to treat a genetic flaw, you need to know which gene(s) to pursue. You must also have a DNA copy of the gene available in your laboratory.

Do you understand the biology of the disorder? To design the best possible approach, you need to learn all you can about how the gene factors into the disorder. For example, which tissues the disorder affects, what role the protein encoded by the gene plays within the cells of that tissue, and exactly how mutations in the gene affect the protein's function.

Will adding a normal copy of the gene fix the problem in the affected tissue? Or could getting rid of the defective gene fix it? Sometimes when a gene is defective, no functional protein is being made from it. In cases like these, adding a functional copy of the gene could correct the problem. But sometimes a defective gene codes for a protein that starts doing something it shouldn't or prevents another protein from doing its job. In order to correct the problem, you would need to get rid of the misbehaving protein.

Can you deliver the gene to cells of the affected tissue? The answer will come from several pieces of information, including the tissue's accessibility and molecular signatures.

APA format: Genetic Science Learning Center (2014, June 22) What is Gene Therapy?. Learn.Genetics. Retrieved May 19, 2015, from http://learn.genetics.utah.edu/content/genetherapy/gtintro/ MLA format: Genetic Science Learning Center. "What is Gene Therapy?." Learn.Genetics 19 May 2015 <http://learn.genetics.utah.edu/content/genetherapy/gtintro/> Chicago format: Genetic Science Learning Center, "What is Gene Therapy?," Learn.Genetics, 22 June 2014, <http://learn.genetics.utah.edu/content/genetherapy/gtintro/> (19 May 2015)

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What is Gene Therapy ? - Learn Genetics

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Tuesday, May 19th, 2015

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Alliance for Cancer Gene Therapy (ACGT) Foundation

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