Transthyretin Gene Expression Research Articles

Human transthyretin gene expression is markedly increased in repair Schwann cells in an in vitro model of hereditary transthyretin amyloidosis.

Hereditary transthyretin (TTR) amyloidosis (ATTRv) is characterized by TTR amyloid deposition in the peripheral nervous system. It remains unknown why variant TTR preferentially deposits in the peripheral nerves and dorsal root ganglia. We previously detected low levels of TTR expression in Schwann cells and established an immortalized Schwann cell line, TgS1, derived from a mouse model of ATTRv amyloidosis expressing the variant TTR gene. In the present study, the expression of TTR and Schwann cell marker genes was investigated in TgS1 cells by quantitative RT-PCR. TTR gene expression was markedly upregulated in TgS1 cells incubated in non-growth medium-Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum. The expression levels of c-Jun, Gdnf and Sox2 were increased, while Mpz was downregulated, suggesting that TgS1 cells exhibit a repair Schwann cell-like phenotype in the non-growth medium. Western blot analysis revealed that TTR protein was produced and secreted by the TgS1 cells. Furthermore, downregulation of Hsf1 with siRNA induced TTR aggregates in the TgS1 cells. These findings indicate that TTR expression is markedly increased in repair Schwann cells, likely to promote axonal regeneration. Therefore, aged dysfunctional repair Schwann cells may cause the deposition of variant TTR aggregates in the nerves of patients with ATTRv.

Perfluorohexanoic acid caused disruption of the hypothalamus-pituitary-thyroid axis in zebrafish larvae

Perfluorohexanoic acid (PFHxA) has been recognized as an alternative to the wide usage of perfluorooctanoate (PFOA) and perfluorooctane sulfonate (PFOS) in the fluoropolymer industry for years. PFHxA has been frequently detected in the environment due to its wide application. However, the ecological safety of PFHxA, especially its toxicological effects on aquatic organisms, remains obscure. In the present study, PFHxA at different concentrations (0, 0.48, 2.4, and 12 mg/L) was added to the culture medium for zebrafish embryo/larval exposure at 96 h postfertilization (hpf). Zebrafish larvae showed a slow body growth trend and changes in thyroid hormone levels (THs) upon PFHxA exposure, indicating the interference effect of PFHxA on fish larval development. Moreover, the transcription levels of genes related to the hypothalamic-pituitary-thyroid (HPT) axis were also analyzed. The gene expression level of thyroid hormone receptor β (trβ) was upregulated in a dose-dependent manner. Exposure to 0.48 mg/L PFHxA increased the expression levels of the thyrotrophic-releasing hormone (trh) and thyroid hormone receptor α (trα). Significant increases in corticotrophin-releasing hormone (crh) and transthyretin (ttr) gene expression were also observed when the zebrafish larvae were treated with 12 mg/L PFHxA, except iodothyronine deiodinases (dio1), which decreased obviously at that point. There were significant declines in the transcription of both thyroid-stimulating hormone β (tshβ) and uridinediphosphate-glucuronosyltransferase (ugt1ab) upon exposure to 2.4 mg/L PFHxA. In addition, PFHxA induced a dose-related inhibitory effect on the transcription of sodium/iodide symporter (nis). Finally, the thyroid status will be destroyed after exposure to PFHxA, thus leading to growth impairment in zebrafish larvae.

Doksycyklina w terapii amyloidozy układowej z zajęciem serca

The leading strategy in the management of systemic amyloidosis is currently focused on reducing the production of amyloid precursor proteins. This approach is based on the use of chemotherapy in light chain amyloidosis (AL amyloidosis) or liver transplantation, or attempts to suppress transthyretin gene expression (TTR) in patients with transthyretin amyloidosis (ATTR). Recently, however, therapies have been increasingly developed to reduce the formation of amyloid deposits from circulating precursors or to eliminate already formed amyloid deposits. This approach includes, in particular, the chronic use of doxycycline, a well-known bacteriostatic antibiotic from the tetracycline group. In preclinical studies it was shown that the anti-amyloidogenic potential of doxycycline in AL amyloidosis depends on interference in the process of amyloidogenesis and the destruction of amyloid deposits. Clinical retrospective studies indicate that doxycycline used with standard chemotherapy improves prognosis in patients with AL amyloidosis with heart involvement, which is the most unfavorable prognostic group, while maintaining a favorable safety profile of therapy. In contrast, in ATTR, doxycycline appears to stabilize the clinical course of the disease. In this paper, we review literature on the role of doxycycline therapy in the treatment of systemic amyloidosis.

Exposure to PFDoA causes disruption of the hypothalamus-pituitary-thyroid axis in zebrafish larvae.

Perfluorododecanoic acid (PFDoA), a kind of perfluorinated carboxylic acid (PFCA) with 12 carbon atoms, has an extensive industrial utilization and is widespread in both wildlife and the water environment, and was reported to have the potential to cause a disruption in the thyroid hormone system homeostasis. In this study, zebrafish embryos/larvae were exposed to different concentrations of PFDoA (0, 0.24, 1.2, 6 mg/L) for 96 h post-fertilization (hpf). PFDoA exposure caused obvious growth restriction connected with the reduced thyroid hormones (THs) contents in zebrafish larvae, strengthening the interference effect on the growth of fish larvae. The transcriptional level of genes within the hypothalamic-pituitary-thyroid (HPT) axis was analyzed. The gene expression levels of thyrotropin-releasing hormone (trh) and corticotrophin-releasing hormone (crh) were upregulated upon exposure to 6 mg/L of PFDoA, and iodothyronine deiodinases (dio2) was upregulated in the 1.2 mg/L PFDoA group. The transcription of thyroglobulin (tg) and thyroid receptor (trβ) were significantly downregulated upon exposure to 1.2 mg/L and 6 mg/L of PFDoA. PFDoA could also decrease the levels of sodium/iodide symporter (nis) and transthyretin (ttr) gene expression in a concentration-dependent manner after exposure. A significant decrease in thyroid-stimulating hormoneβ (tshβ), uridinediphosphate-glucuronosyltransferase (ugt1ab) and thyroid receptor (trα) gene expression were observed at 6 mg/L PFDoA exposure. Upregulation and downregulation of iodothyronine deiodinases (dio1) gene expression were observed upon the treatment of 1.2 mg/L and 6 mg/L PFDoA, respectively. All the data demonstrated that gene expression in the HPT axis altered after different PFDoA treatment and the potential mechanisms of the disruption of thyroid status could occur at several steps in the process of synthesis, regulation, and action of thyroid hormones.

Transthyretin gene mutation and expression in patients with familial vitreous amyloidosis

Objective To observe the transthyretin (TTR) gene mutation, protein and mRNA expression in patients with familial vitreous amyloidosis. Methods Subjects were divided into three groups: (1) illness group: seven patients with familial vitreous amyloidosis. (2) No-illness group: 9 unaffected family members. (3) Control group: 9 healthy individuals in same area. Subjects' peripheral venous blood were collected and DNA were extracted, 4 exons of TTR gene were amplified by reverse transcription polymerase chain reaction(RT-PCR), the gene fragments were sequencing by the fluorescence labelling method. Serum TTR protein expression was detected by Western blot, and TTR mRNA in leukocyte was assayed by RT-PCR. Results 4 exons of TTR gene of all samples were amplified, and DNA sequencing data showed that 7 patients and 3 subjects DNA from unaffected family members had mutated in the 3rd exon of 107th base, changing from G to C. Heterozygous mutation occurred in codon of the 83th amino acid in exon 3, namely, Gly83Arg, resulted in the change of GGC to CGC. The protein and mRNA expression of TTR was lower in illness group than no-illness group and control groups(P<0.05). Compared with control group, TTR mRNA expression in unaffected family members groups was significant decreased(P<0.05). Conclusion Heterozygous mutation occurred in codon of the 83th amino acid in exon 3, namely Gly83Arg, and suggested that Gly83Arg is connected with the change of TTR mRNA and protein expression. Key words: Amyloidosis, familial/genetics; Thyroid hormones; DNA mutational analysis

Correction for Li et al., Soluble amyloid precursor protein (APP) regulates transthyretin and Klotho gene expression without rescuing the essential function of APP

Correction for Li et al., Soluble amyloid precursor protein (APP) regulates <i>transthyretin</i> and <i>Klotho</i> gene expression without rescuing the essential function of APP

Reevaluation of transthyretin gene expression in the peripheral nervous system

Reevaluation of transthyretin gene expression in the peripheral nervous system

Hepatocyte nuclear factor‐4α interacts with other hepatocyte nuclear factors in regulating transthyretin gene expression

Transthyretin is a negative acute phase protein whose serum level decreases during the acute phase response. Transthyretin gene expression in the liver is regulated at the transcriptional level, and is controlled by hepatocyte nuclear factor (HNF)-4α and other HNFs. The site-directed mutagenesis of HNF-4, HNF-1, HNF-3 and HNF-6 binding sites in the transthyretin proximal promoter dramatically decreases transthyretin promoter activity. Interestingly, the mutation of the HNF-4 binding site not only abolishes the response to HNF-4α, but also reduces significantly the response to other HNFs. However, mutation of the HNF-4 binding site merely affects the specific binding of HNF-4α, but not other HNFs, suggesting that an intact HNF-4 binding site not only provides a platform for specific interaction with HNF-4α, but also facilitates the interaction of HNF-4α with other HNFs. In a cytokine-induced acute phase response cell culture model, we observed a significant reduction in the binding of HNF-4α, HNF-1α, HNF-3β and HNF-6α to the transthyretin promoter, which correlates with a decrease in transthyretin expression after injury. These findings provide new insights into the mechanism of the negative transcriptional regulation of the transthyretin gene after injury caused by a decrease in the binding of HNFs and a modulation in their coordinated interactions.

Evolutionary changes to transthyretin: evolution of transthyretin biosynthesis

Thyroid hormones are involved in growth and development, particularly of the brain. Thus, it is imperative that these hormones get from their site of synthesis to their sites of action throughout the body and the brain. This role is fulfilled by thyroid hormone distributor proteins. Of particular interest is transthyretin, which in mammals is synthesized in the liver, choroid plexus, meninges, retinal and ciliary pigment epithelia, visceral yolk sac, placenta, pancreas and intestines, whereas the other thyroid hormone distributor proteins are synthesized only in the liver. Transthyretin is synthesized by all classes of vertebrates; however, the tissue specificity of transthyretin gene expression varies widely between classes. This review summarizes what is currently known about the evolution of transthyretin synthesis in vertebrates and presents hypotheses regarding tissue-specific synthesis of transthyretin in each vertebrate class.

Evolutionary changes to transthyretin

Thyroid hormones (THs) are key regulators of growth, differentiation and metabolism of vertebrates. They are soluble in lipid membranes, and binding to specific proteins in plasma could thereby prevent their loss from the circulation. Albumin, transthyretin and thyroxine-binding globulin (TBG) are the major distributors of THs in the plasma of larger mammals. Of these TH-binding proteins, transthyretin is one of the most interesting. In this minireview series, Prapunpoj and Leelawatwattana review the work carried out to elucidate the 3D structures and functions of transthyretins from several vertebrates. During evolution, the amino acid residues involved in TH binding have not been altered. However, pronounced changes have occurred in the Nand, to a lesser extent, in the C-terminal regions. The question arose as to whether these alterations influence the function of transthyretin and this point is discussed in the review. Transthyretin has versatile functions and these are suggested to depend on its binding affinities mainly to THs and to retinol-binding protein (RBP). Transthyretin works in a ‘network system’ with albumin and TBG in distributing THs from the thyroid gland to target tissues. In this network, a deficiency in one component can be compensated for by the other compounds, which has meant that any specific function of transthyretin could not be revealed directly by means of gene knockout. Analysis of the specific sites of synthesis of transthyretin and the onset of transthyretin gene expression in tissues during vertebrate evolution is one effective way of elucidating the putative functions of transthyretin. This, and hypotheses regarding tissue-specific synthesis of transthyretin in each vertebrate class, are summarized and discussed by Richardson. The evolutionary expression of a gene in specific tissues can indicate the importance and functions of the gene product. The developmentally regulated geneexpression profile of the transthyretin and other TH carriers in vertebrates, reviewed by Yamauchi and Ishihara, indicates how the TH distributor network in the extracellular space developed during evolution and how this bloodstream network evolved with functional redundancy. A correlation of the appearance of the network with the increased requirement of THs for TH-dependent tissue remodeling and ⁄or increased metabolism in TH-target tissues with the acquisition of homeothermy, has been proposed. The primary structures of transthyretins from fish and amphibians are similar to those from humans, suggesting that there has been little change since the divergence of the vertebrates. Multiple genomes have been identified and characterized via nucleotide sequencing coupled with sophisticated pairwise sequence-alignment search tools. These have also enabled a search for a transthyretin homologue, transthyretin-like protein (TLP), among nonvertebrate organisms, as summarized by Hennebry. The distribution of the TLP genes in nature, and the structure, function and evolution of the TLP proteins from various organisms, are discussed. This minireview series illustrates what we know about the evolution of transthyretin. The knowledge obtained not only provides insight into the importance and functional aspects of transthyretin, but also points the way to future research directions relating to this versatile molecule.

A Diet Enriched with the Omega-3 Fatty Acid Docosahexaenoic Acid Reduces Amyloid Burden in an Aged Alzheimer Mouse Model

Epidemiological studies suggest that increased intake of the omega-3 (n-3) polyunsaturated fatty acid (PUFA) docosahexaenoic acid (DHA) is associated with reduced risk of Alzheimer's disease (AD). DHA levels are lower in serum and brains of AD patients, which could result from low dietary intake and/or PUFA oxidation. Because effects of DHA on Alzheimer pathogenesis, particularly on amyloidosis, are unknown, we used the APPsw (Tg2576) transgenic mouse model to evaluate the impact of dietary DHA on amyloid precursor protein (APP) processing and amyloid burden. Aged animals (17-19 months old) were placed in one of three groups until 22.5 months of age: control (0.09% DHA), low-DHA (0%), or high-DHA (0.6%) chow. beta-Amyloid (Abeta) ELISA of the detergent-insoluble extract of cortical homogenates showed that DHA-enriched diets significantly reduced total Abeta by >70% when compared with low-DHA or control chow diets. Dietary DHA also decreased Abeta42 levels below those seen with control chow. Image analysis of brain sections with an antibody against Abeta (amino acids 1-13) revealed that overall plaque burden was significantly reduced by 40.3%, with the largest reductions (40-50%) in the hippocampus and parietal cortex. DHA modulated APP processing by decreasing both alpha- and beta-APP C-terminal fragment products and full-length APP. BACE1 (beta-secretase activity of the beta-site APP-cleaving enzyme), ApoE (apolipoprotein E), and transthyretin gene expression were unchanged with the high-DHA diet. Together, these results suggest that dietary DHA could be protective against beta-amyloid production, accumulation, and potential downstream toxicity.

The evolution of the thyroid hormone distributor protein transthyretin in the order insectivora, class mammalia.

Thyroid hormones are involved in the regulation of growth and metabolism in all vertebrates. Transthyretin is one of the extracellular proteins with high affinity for thyroid hormones which determine the partitioning of these hormones between extracellular compartments and intracellular lipids. During vertebrate evolution, both the tissue pattern of expression and the structure of the gene for transthyretin underwent characteristic changes. The purpose of this study was to characterize the position of Insectivora in the evolution of transthyretin in eutherians, a subclass of Mammalia. Transthyretin was identified by thyroxine binding and Western analysis in the blood of adult shrews, hedgehogs, and moles. Transthyretin is synthesized in the liver and secreted into the bloodstream, similar to the situation for other adult eutherians, birds, and diprotodont marsupials, but different from that for adult fish, amphibians, reptiles, monotremes, and Australian polyprotodont marsupials. For the characterization of the structure of the gene and the processing of mRNA for transthyretin, cDNA libraries were prepared from RNA from hedgehog and shrew livers, and full-length cDNA clones were isolated and sequenced. Sections of genomic DNA in the regions coding for the splice sites between exons 1 and 2 were synthesized by polymerase chain reaction and sequenced. The location of splicing was deduced from comparison of genomic with cDNA nucleotide sequences. Changes in the nucleotide sequence of the transthyretin gene during evolution are most pronounced in the region coding for the N-terminal region of the protein. Both the derived overall amino sequences and the N-terminal regions of the transthyretins in Insectivora were found to be very similar to those in other eutherians but differed from those found in marsupials, birds, reptiles, amphibians, and fish. Also, the pattern of transthyretin precursor mRNA splicing in Insectivora was more similar to that in other eutherians than to that in marsupials, reptiles, and birds. Thus, in contrast to the marsupials, with a different pattern of transthyretin gene expression in the evolutionarily "older" polyprotodonts compared with the evolutionarily "younger" diprotodonts, no separate lineages of transthyretin evolution could be identified in eutherians. We conclude that transthyretin gene expression in the liver of adult eutherians probably appeared before the branching of the lineages leading to modern eutherian species.

Evolution of transthyretin gene expression in the liver of Didelphis virginiana and other American marsupials

The occurrence of the thyroid hormone-binding plasma protein transthyretin in the bloodstream was investigated for four American marsupial species. Serum samples were analyzed by incubation with radioactive T4, followed by electrophoresis, then autoradiography, and Western blotting. Transthyretin was found in serum from Monodelphis domestica, Didelphis virginiana, Caluromys lanatus, and Dromiciops australis. For unambiguous identification, transthyretin from D, virginiana was purified from serum and its N-terminal amino acid sequence was determined. The obtained results suggest that the initiation of transthyretin gene expression in the liver of marsupials occurred independently in several lineages of American marsupials, all of which are at the ends of phylogenetic branches. The expression of the transthyretin gene in the liver of the American polyprotodont marsupials contrasts with the lack of transthyretin gene expression in the liver of all 22 previously investigated Australian Polyprotodonta.

Evolution of marsupial and other vertebrate thyroxine-binding plasma proteins

Binding of radioactive thyroxine to proteins in the plasma of vertebrates was studied by electrophoresis followed by autoradiography. Albumin was found to be a thyroxine carrier in the blood of all studied fish, amphibians, reptiles, monotremes, marsupials, eutherians (placental mammals), and birds. Thyroxine binding to transthyretin was detected in the blood of eutherians, diprotodont marsupials, and birds, but not in blood from fish, toads, reptiles, monotremes, and Australian polyprotodont marsupials. Globulins binding thyroxine were only observed in the plasma of some mammals. Apparently, albumin is the phylogenetically oldest thyroxine carrier in vertebrate blood. Transthyretin gene expression in the liver developed in parallel, and independently, in the evolutionary lineages leading to eutherians, to diprotodont marsupials, and to birds. In contrast, high transthyretin mRNA levels, strong synthesis, and secretion of transthyretin in choroid plexus from reptiles and birds indicate that transthyretin gene expression in the choroid plexus evolved much earlier than in the liver, probably at the stage of the stem reptiles. NH2-terminal sequence analysis suggests a change of transthyretin pre-mRNA splicing during evolution.

Transthyretin expression evolved more recently in liver than in brain

1. 1. Transthyretin was found to be synthesized and secreted by choroid plexus from rats, echidnas, and lizards, but not toads. 2. 2. Transthyretin was observed in blood from placental mammals, birds, and marsupials, but not reptiles and monotremes. 3. 3. The obtained data suggest that transthyretin synthesis by the liver evolved independently in the lineage leading to the placental mammals and marsupials and in that leading to the birds. 4. 4. It is proposed that transthyretin gene expression in mammalian liver appeared about 200 million years later than its first occurrence in the choroid plexus of the stem reptiles.

The Expression of the Transthyretin Gene in Liver Evolved during the Radiation of Diprotodont Marsupials in Australia

Thyroid hormone-binding proteins in blood plasma were identified in 28 different marsupial species by their capacity to bind radioactive thyroxine. All species contained albumin. Transthyretin was not found in the blood from any of 12 polyprotodont marsupial species, but was abundant in the blood from all of 16 diprotodont marsupial species investigated. Transthyretin mRNA was absent from the liver of the stripe-faced dunnart, a polyprotodont marsupial, but abundant in the liver of the diprotodont grey kangaroo. Diprotodont marsupials probably evolved in Australia from polyprotodont marsupials after their transantarctic migration from South America, about 40 million years ago. It is suggested that hepatic transthyretin expression evolved in marsupials during the radiation of herbivorous, diprotodont species in Australia. The earlier appearance of transthyretin gene expression in the choroid plexus of the stem reptiles, about 300 million years ago, contrasts with hepatic transthyretin synthesis, a relatively late evolutionary event, occurring independently in at least three lineages.

Specific repression of transthyretin gene expression in rat liver by a peroxisome proliferator clofibrate

The effect of clofibrate, a hypolipidemic drug and a potent peroxisome proliferator, on expression of a nonperoxisomal transthyretin (prealbumin) gene was studied using rats fed clofibrate for various periods. Upon feeding a diet containing clofibrate, the level of transthyretin mRNA was down-regulated, reaching 20% of the control level, in an almost reciprocal time course to that of increases in the levels of peroxisomal mRNAs. Studies on expression of other steroid hormone regulated genes suggest that clofibrate may down-regulate several but not all types of steroid hormone regulated mRNA expression.

Transthyretin (prealbumin) gene expression in choroid plexus is strongly conserved during evolution of vertebrates

1. 1. The major protein synthesized and secreted by the choroid plexus from mammals, birds, reptiles and probably amphibians is similar in subunit structure to transthyretin. 2. 2. In mammals and birds the proportion of transthyretin mRNA is much higher in choroid plexus RNA than in liver RNA. No transthyretin mRNA is found in brain outside the choroid plexus. 3. 3. Transthyretin-like protein, such as that secreted by the choroid plexus, was not detected in amphibian serum and was present in very low levels in reptile serum. 4. 4. It is proposed that transthyretin synthesis and secretion arose earlier in evolution in the choroid plexus than in the liver.

Ontogenesis of transthyretin gene expression in chicken choroid plexus and liver

1. Chicken liver transthyretin cDNA hybridizes strongly with choroid plexus transthyretin mRNA from chickens, pigeons, quails and ducks. 2. In the chicken at hatching the choroid plexus has reached 70%, total brain 30%, and liver 5.8% of their organ masses in adults. 3. The proportion of transthyretin mRNA in total RNA is 0.45-times the adult value in the choroid plexus of the chicken at hatching. 4. In the liver at hatching, the proportion of transthyretin mRNA in total RNA is 1.1-times the value in adult chickens. 5. The pattern of maturation of transthyretin gene expression in chicken liver is comparable to that in precocial, but differs from that in altricial mammals.

Similarities in transthyretin gene expression and differences in transcription factors: liver and yolk sac compared to choroid plexus.

The serum thyroxine-binding protein, transthyretin (TTR), is made by hepatocytes and by choroid plexus epithelium in adults and by yolk sac cells in embryogenesis. Four hepatocyte nuclear factors (HNF-1, -3, and -4 and C/EBP) that are present in liver but not in most other adult tissues bind DNA sites in the TTR gene that are sufficient to direct transgenic expression. Three of these proteins were also found in yolk sac cells, which also express the transgene. A limited transgenic construct is not active in the choroid plexus and a TTR-producing choroid plexus tumor lacks three of the liver-enriched DNA-binding proteins. We conclude that cell-specific expression of TTR is regulated at least in part by the differential cellular distribution of positive-acting transcription factors.