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In collaboration with an international research team, University of Copenhagen researchers have for the first time mapped telomerase, an enzyme which has a kind of rejuvenating effect on normal cell ageing. The findings have just been published in Nature Genetics and are a step forward in the fight against cancer.
Mapping the cellular fountain of youth – telomerase. This is one of the results of a major research project involving more than 1,000 researchers worldwide, four years of hard work, DKK 55 million from the EU and blood samples from more than 200,000 people. This is the largest collaboration project ever to be conducted within cancer genetics.
Stig E. Bojesen, a researcher at the Faculty of Health and Medicial Sciences, University of Copenhagen, and staff specialist at the Department of Clinical Biochemistry, Copenhagen University Hospital, Herlev, has headed the efforts to map telomerase – an enzyme capable of creating new ends on cellular chromosomes, the so-called telomeres. In other words, a kind of cellular fountain of youth.
“We have discovered that differences in the telomeric gene are associated both with the risk of various cancers and with the length of the telomeres. The surprising finding was that the variants that caused the diseases were not the same as the ones which changed the length of the telomeres. This suggests that telomerase plays a far more complex role than previously assumed,” says Stig E. Bojesen.
The mapping of telomerase is an important discovery, because telomerase is one of the very basic enzymes in cell biology. It relengthens the telomeres so that they get the same length as before embarking on cell division.
“The mapping of telomerase may, among other things, boost our knowledge of cancers and their treatment, and with the new findings the genetic correlation between cancer and telomere length has been thoroughly illustrated for the first time,” says Stig E. Bojesen.
Telomeres a cellular ‘multi-ride ticket’
The human body consists of 50,000,000,000,000 or fifty trillion cells, and each cell has 46 chromosomes which are the structures in the nucleus containing our hereditary material, the DNA. The ends of all chromosomes are protected by so-called telomeres. The telomeres serve to protect the chromosomes in much the same way as the plastic sheath on the end of a shoelace. But each time a cell divides, the telomeres become a little bit shorter and eventually end up being too short to protect the chromosomes.
Popularly speaking, each cell has a multi-ride ticket, and each time the cell divides, the telomeres (the chromosome ends) will use up one ride. Once there are no more rides left, the cell will not divide any more, and will, so to speak, retire. But some special cells in the body can activate telomerase, which again can elongate the telomeres.
Sex cells, or other stem cells which must be able to divide more than normal cells, have this feature. Unfortunately, cancer cells have discovered the trick, and it is known that they also produce telomerase and thus keep themselves artificially young. The telomerase gene therefore plays an important role in cancer biology, and it is precisely by identifying cancer genes that the researchers imagine that you can improve the identification rate and the treatment.
“A gene is like a country. As you map it, you can see what is going on in the various cities. One of the cities in what could be called Telomerase Land determines whether you develop breast cancer or ovarian cancer, while other parts of the gene determine the length of the telomeres. Mapping telomerase is therefore an important step towards being able to predict the risk of developing different cancers. In summary, our findings are very surprising and point in many directions. But as is the case with all good research, our work provides many answers but leaves even more questions,” says Stig E. Bojesen.
At a glance
Chromosome ends are capped by telomeres, which protect them from inappropriate DNA repair and maintain genomic integrity1. Telomeres consist of structural proteins2 combined with many hundreds of hexanucleotide DNA repeats3, 4, which are progressively shortened by normal cell division5, 6, 7. Shortening restricts the proliferation of normal somatic cells but not cancer cells, which can maintain long telomeres, usually via telomerase8, 9, 10, and may divide indefinitely. The TERT gene at 5p15.33 (NCBI gene 7015) encodes the catalytic subunit of telomerase reverse transcriptase, a key component of telomerase. Germline mutations in TERT cause dyskeratosis congenita, a cancer susceptibility disorder characterized by exceedingly short telomeres11. Although up to 80% of the variation of telomere length is estimated to be due to heritable factors12, 13, association studies of TERT SNPs and differences in leukocyte telomere length have so far been inconclusive14, 15, 16, 17. Furthermore, it is unclear whether telomere length, measured in leukocyte DNA, is predictive of cancer risk: retrospective studies report that cancer patients after diagnosis have shorter telomeres than unaffected controls18, 19, 20, 21, but prospective studies with DNA taken before diagnosis have been inconclusive19, 22, 23. SNPs at 5p15.33 are reported to be associated with risks of several human cancers14, 15, 16, 24, 25, 26, 27, 28, 29, 30, 31, 32, including certain subtypes of both ovarian33 and breast34 cancers.
Resulting from a common interest, members of each of the constituent consortia in the Collaborative Oncological Gene-environment Study (COGS) nominated SNPs surrounding the TERT locus for inclusion on a genotyping array. Consequently, the iCOGS array design included a combination of individual TERT gene candidate SNPs, as well as a more comprehensive set to fine-scale map the entire locus, for shared use by all consortia. This study had three aims: to assess SNPs across the TERT locus for all detectable associations with mean telomere length and breast and ovarian cancer subtypes; to fine-scale map this locus to identify potentially causal variants for the observed associations; and to evaluate the functional effects of the strongest candidate causative variants.
One hundred and ten SNPs at the 5p15.33 locus (Build 37 positions 1,227,693–1,361,969) passed quality control tests in 103,991 breast cancer cases and controls from 52 Breast Cancer Association Consortium (BCAC) studies, of which 41 studies (89,050 individuals) were of European ancestry, 9 were of Asian ancestry (12,893 individuals) and 2 were of African-American ancestry (2,048 individuals). The same 110 SNPs passed quality control tests in 11,705 BRCA1 mutation carriers of European ancestry, recruited by 45 studies from the Consortium of Investigators of Modifiers of BRCA1 and BRCA2 (CIMBA), and 108 SNPs passed quality control tests in 44,308 ovarian cancer cases and controls from 43 Ovarian Cancer Association Consortium (OCAC) studies. For OCAC, analysis was confined to the 39,774 participants of European ancestry, of whom 8,371 cases had invasive epithelial ovarian neoplasia and 986 had serous low-malignant-potential (LMP) neoplasia. For all study participants, genotype imputation, using the 110 genotyped SNPs together with the January 2012 release of the 1000 Genomes Project35, 36, 37, 38, was used to increase coverage to ~480 SNPs (imputation r2 > 0.3; minor allele frequency (MAF) > 0.02) for each phenotype. Telomere length was initially measured in control subjects from two BCAC studies (Studies of Epidemiology And Risk factors in Cancer Heredity (SEARCH) and the Copenhagen City Heart Study (CCHS); combined n = 15,567) (Supplementary Note).
Manhattan plots are shown of the genotyped and well-imputed SNPs for the seven phenotypes analyzed, including mean telomere length (Fig. 1a), overall breast cancer risk (Fig. 1b), breast cancer in BRCA1 mutation carriers (Fig. 1c), ER-negative breast cancer (Fig. 1d), ER-positive breast cancer (Fig. 1e), serous LMP ovarian cancer (Fig. 1f) and serous invasive ovarian cancer (Fig. 1g). Conditional analyses within each of these phenotypes identified multiple independent SNP associations each for telomere length, overall breast cancer risk, ER-negative breast cancer and breast cancer in BRCA1 mutation carriers but only one peak each for ER-positive breast cancer, serous LMP ovarian cancer and invasive ovarian cancer (Table 1). Full results of all these SNP analyses are given in Supplementary Tables 1–3. All associations are consistent with a log-additive model.
Associations with telomere length
SNPs in two distinct regions (hereafter denoted peaks 1 and 2) were strongly associated with telomere length (Fig. 1a, Tables 1 and 2 and Supplementary Fig. 1a). Imputed SNP rs7705526 (peak 2, position 1,285,974, TERT intron 2) had the largest effect, with a change in relative telomere length of 1.026-fold per allele (95% confidence interval (CI) = 1.019–1.033; P = 2.3 × 10−14; conditional P = 2.5 × 10−11). We confirmed this finding in an additional 20,512 women and 17,645 men from a third study (CGPS) genotyped for rs7726159 (the best directly genotyped SNP; r2 = 0.83 with rs7705526). From a joint analysis of all 53,724 individuals, the change in relative telomere length was 1.020-fold per allele (95% CI = 1.016–1.023; P = 7.5 × 10−28). A second, independent association was observed with rs2736108 (peak 1, position 1,297,488, TERT promoter) with a per-allele change in relative telomere length of 1.017-fold (95% CI = 1.010–1.024; P = 5.8 × 10−7; conditional P = 4.0 × 10−4) (Fig. 1a, Tables 1 and 2 and Supplementary Fig. 1a). SNPs rs7705526 and rs2736108 were only weakly correlated (r2 = 0.04 in Europeans). Weak associations between peak 3 SNPs and telomere length became nonsignificant after adjustment for peak 2 SNP rs7705526 (data not shown).
Associations with breast cancer risk
We identified SNPs associated with breast cancer risk (P < 1 × 10−4) in three distinct regions in subjects from the BCAC studies and in two regions in CIMBA BRCA1 mutation carriers. No significant (P < 1 × 10−4) evidence for heterogeneity among odds ratios (ORs) or hazard ratios (HRs) between studies for any of the top SNPs was observed (Supplementary Fig. 2). The strongest association with overall breast cancer risk in BCAC was with peak 1 SNP rs3215401 (Fig. 1b, Tables 1 and 2 and Supplementary Fig. 1b). There was also good evidence for an association with SNPs in peak 2 and weaker evidence that an additional SNP, outside the three main association peaks, was independently associated with breast cancer risk (Table 1 and Supplementary Table 1). The most strongly associated SNPs in BRCA1 mutation carriers were located in introns 2–4 (hereafter denoted peak 3), including rs10069690 (Fig. 1c, Tables 1 and 2 and Supplementary Fig. 2c) and rs2242652 (correlation with rs10069690, r2 = 0.70). The latter SNP also showed the strongest association with ER-negative breast cancer in BCAC (Fig. 1d, Tables 1 and 2 and Supplementary Fig. 1d) but showed little evidence of association with ER-positive breast cancer (Table 2). Stepwise regression analysis in CIMBA studies indicated two independent associations with breast cancer risk in BRCA1 mutation carriers (conditional P = 5 × 10−5 for rs2736108 in peak 1 and P = 4.8 × 10−13 for rs10069690 in peak 3). A very similar pattern was observed for ER-negative breast cancer in BCAC (conditional P = 6 × 10−6 for rs3215401 in peak 1 and P = 4.3 × 10−9 for rs2242652 in peak 3; Table 1).
The most strongly associated SNP with ER-positive breast cancer was rs2736107 in peak 1 (Fig. 1e, Tables 1 and 2 and Supplementary Fig. 3e). Weak associations between the key SNPs and risk for BRCA2 mutation carriers were also observed, but the sample size was too small to draw definitive conclusions (data not shown).
Associations with ovarian cancer risk
The strongest association observed for risk of LMP ovarian cancer was with peak 2 SNP rs7705526, and this was the only SNP retained in the stepwise regression analysis (Fig. 1f, Tables 1 and 2 and Supplementary Fig. 1f). The strongest observed association for serous invasive ovarian cancer was with peak 3 SNP rs10069690 (Fig. 1g, Tables 1 and 2 and Supplementary Fig. 1g). No other independent association was observed for serous invasive ovarian cancer (Table 1). We also analyzed SNP associations with endometrioid, mucinous, clear-cell invasive and mucinous LMP ovarian cancers but found no associations at P < 1 × 10−4 (Supplementary Table 4). We attempted analysis of invasive serous ovarian cancer stratified by grade, but, again, statistical power was low (Supplementary Fig. 3).
Three main peaks of association within the TERT locus
The above results indicate that the majority of observed associations with all seven tested phenotypes fall into association peaks 1–3. Correlated SNPs in the TERT promoter (peak 1) were associated with telomere length, ER-positive breast cancer, ER-negative breast cancer and breast cancer in BRCA1 mutation carriers. SNPs in peak 2, spanning TERT introns 2–4, were independently associated with telomere length, overall breast cancer risk and serous LMP ovarian cancer. SNPs in peak 3, also spanning TERT introns 2–4, showed strong associations with ER-negative breast cancer, breast cancer risk for BRCA1 mutation carriers and serous invasive ovarian cancer but not with telomere length (Tables 1 and 2). Although peaks 2 and 3 overlap physically, they define distinct sets of SNPs that are only partially correlated (for example, correlation between rs10069690 and rs7705526 was weak, r2 = 0.33; Fig. 2). Some SNP-phenotype associations in peak 2 were clearly weaker than those in peak 3 (for example, with ER-negative breast cancer) and became nonsignificant after adjustment for SNP rs2242652 in peak 3. Conversely, the associations with telomere length and serous LMP ovarian cancer were stronger for SNPs in peak 2, indicating that the associations in peaks 2 and 3 are not being driven by the same causal variants.
The strongest candidates for causation within each peak were identified by computing likelihood ratios; the SNPs listed in Tables 1 and 2 are those that cannot be excluded from being causal candidates at a likelihood ratio of >1:100 fold compared to the top hit in the peak. The statistical power to exclude SNPs differed between phenotypes: in peak 1, all but seven SNPs could be excluded from being causal for relative telomere length, breast cancer risk in BRCA1 mutation carriers and ER-negative breast cancer risk, but an additional SNP could be excluded for ER-positive breast cancer risk (Table 2). In peak 2, the greatest power was for the telomere length phenotype, where all but three SNPs could be excluded, whereas five or six remained for cancer risk. For peak 3, three putative causal SNPs remained for ER-negative breast cancer risk, two for serous invasive ovarian cancer risk and just one for breast cancer risk in BRCA1 mutation carriers. Results in each peak are compatible with a single causative variant being responsible for the multiple phenotype associations (notably, in peak 3, SNPs rs2242652 and rs10069690 were equally compatible with being the single causal variant). However, the possibilities of different causal variants being responsible for different phenotypes or of the associations being due to haplotype effects cannot be ruled out.
Asian and African-American studies
We tested all SNPs (n = 341) with MAF > 0.02 and imputation r2 > 0.3 for association with breast cancer in the 9 BCAC Asian studies (comprising 6,269 cases and 6,624 controls) for association, but none reached formal levels of significance. Furthermore, none of the top SNPs in individuals of European ancestry showed more than borderline levels of significance in Asians (Supplementary Table 5). Peak 3 SNP rs10069690 was directly genotyped in 2 BCAC African-American studies (1,116 cases and 932 controls), as well as in the above-mentioned Asian studies, and had estimated effects on ER-negative breast cancer similar to those in European populations (per-allele OR = 1.19, 95% CI = 1.06–1.31, P = 0.009 in African- Americans and OR = 1.09, 95% CI = 1.00–1.19, P = 0.07 in Asian women). Within OCAC, there were too few women of Asian and African ancestry to draw meaningful conclusions (Supplementary Table 6).
Analysis of Encyclopedia of DNA Elements (ENCODE) data39 showed no evidence of regulatory elements or open chromatin coinciding with any risk-associated SNPs in normal breast epithelial cells or the other represented tissues (Supplementary Fig. 4). Data for ovarian tissues are not included in ENCODE. We therefore performed site-specific formaldehyde-assisted isolation of regulatory elements (FAIRE)40 in ovarian cancer precursor tissues to identify regulatory elements in a 1 Mb region centered on peak 3. In fallopian tube secretory and ovarian surface epithelial cells, we detected FAIRE peaks coinciding with the CLPTM1L promoter but not the TERT promoter (Supplementary Fig. 4). In silico analyses additionally indicated that TERT introns 4 and 5 (within and beyond peak 3) contained regions showing regulatory potential and vertebrate sequence conservation41. We performed site-specific FAIRE analyses on a ~1 kb region centered on the peak 3 SNP rs10069690 in normal tissue samples from breast reduction mammoplasty (n = 4), ovarian cancer precursor tissues (n = 4) and ovarian cancer cell lines (n = 4). Breast cells from each woman were sorted into four enriched fractions on the basis of differential expression of cell surface markers42 (myoepithelial/stem, luminal progenitor, mature luminal and stromal cells), and assays were performed on each fraction (Fig. 3). Chromatin was in a closed configuration in all ovarian, breast luminal progenitor and mature luminal cell fractions. However, in two of four stromal cell fractions, we detected ~600 bp of open chromatin of varying amplitude, covering the position of SNP rs10069690 but not of rs2242652, and, in three of four myoepithelial/stem cell fractions, we detected ~800 bp of open chromatin, covering the positions of both SNPs rs10069690 and rs2242652.
Luciferase reporter assays
The regulatory capabilities of the DNA in each of the three peaks and the effects of most of the strongest candidate causative variants in each one were examined in luciferase reporter assays, using a construct containing 3,915 bp of the TERT promoter sequence43. The effects of peak 1 TERT promoter variants were examined via five haplotype constructs differing at rs2736107, rs2736108 and rs2736109 (ref. 25) (Fig. 4a): one with all three major alleles (wild-type TERT), another with all three minor alleles and three each with a single minor allele of the SNPs. Relative promoter activity was determined in ER-positive (MCF7) and ER-negative (MDA-MB-468) breast cancer cell lines and in an ovarian cancer cell line (A2780). The construct containing all three minor alleles consistently generated the lowest luciferase signals, close to baseline. To determine whether the risk-associated variants in peaks 2 and 3 fell within putative cis-acting regulatory elements (PREs), we cloned ~3 kb of sequence surrounding each SNP. Constructs of PRE-A (peak 2) had no significant effect on the activity of either the wild-type (TERTwt) promoter or the promoter with three minor alleles (TERTh) (Fig. 4a). However, inclusion of the minor allele of rs7705526 resulted in ~30% higher TERT promoter activity in all three cell lines, suggesting that it can act as a transcriptional enhancer. Higher promoter activity was also observed with this construct in A2780 ovarian cancer cells but not in the two breast cancer cell lines. Constructs of PRE-B (peak 3) consistently acted as strong transcriptional silencers, leading to 40–50% lower activity, specifically in constructs containing the wild-type TERT promoter. Notably, inclusion of the minor allele of rs2242652 in PRE-B constructs decreased relative wild-type TERT promoter activity by a further ~20% compared to the silencer containing the major allele, but the minor allele of the highly correlated SNP rs10069690 did not generate this effect (Fig. 4a).
Alternative splicing of TERT
Several alternatively spliced variants of TERT have been found to affect telomerase activity44, 45. To determine the role of PRE-B (peak 3) SNPs in TERT alternative splicing, we inserted intron 4 sequence into a full-length TERT cDNA mini-gene construct and confirmed accurate splicing. Cancer risk–associated alleles for rs10069690 and rs2242652 were generated individually and in combination within the mini-gene. RT-PCR, using primers spanning intron 4, showed that all SNP permutations in all cell lines produced comparable levels of both wild-type transcript and an INS1 alternatively spliced variant, which includes the first 38 bp of TERT intron 4 (refs. 46, 47) (Supplementary Fig. 5a). We also identified a new splice variant of TERT, specifically associated with the minor allele of rs10069690 (termed INS1b; Supplementary Fig. 5a). Sequence analysis confirmed that INS1b includes the first 480 bp of intron 4 and results from the use of an alternative splice donor created by the minor allele of rs10069690 (ref. 48). INS1b has a premature stop codon 16 amino acids into intron 4 and is predicted to generate a severely truncated protein product, which is likely to affect telomerase activity (Supplementary Fig. 5b).
Gene expression and methylation analyses in ovarian tissue
We used The Cancer Genome Atlas (TCGA)49 data to examine gene expression of the 11 protein-coding genes and 1 microRNA (MIR4457) located within 1 Mb of peak 3 SNP rs10069690. Most genes showed higher expression in ovarian tumors compared with normal tissues (Supplementary Fig. 4 and Supplementary Table 7). We observed no association between rs10069690 and the expression levels of any of the genes in any of the cells tested (Supplementary Fig. 5 and Supplementary Tables 7 and 8). There was some evidence of association between rs10069690 and tumor methylation with probes cg23827991 (TERT CpG island, P = 1.3 × 10−6) and cg06550200 (CLPTM1L, P = 6.9 × 10−4) out of the 935 probes tested. Both regions showed lower methylation with the minor, cancer risk–associated allele (Supplementary Table 9), but this did not correlate with changes in expression.
Our comprehensive examination of the TERT locus has answered some long-standing questions and raised several new ones. We have identified two independent regions associated with telomere length in leukocyte DNA; these provide definitive evidence for genetic control of telomere length by common TERT variants. For rs2736108, the most significant SNP in promoter peak 1, the minor allele is associated with a 1.7% increase in telomere length. This is equal to a telomere length change of ~60 bp, which, because telomere length decreases by approximately 19 bp per year50, is equivalent in magnitude to an age difference of 3.1 years. We estimate that rs2736108 explains 0.08% of the variance in telomere length in men and 0.06% of the variation in women. SNPs in peak 2 have a stronger effect on telomere length, with each additional A (minor) allele of rs7705526 associated with a 2.6% increase. This is equal to a ~90 bp change in telomere length and, correspondingly, to 4.7 years of age. We estimate that rs7705526 explains 0.31% of the variance in telomere length in men and 0.16% of the variance in women. The only other reported associations with telomere length reaching genome-wide significance involve TERC-locus SNP rs1269304 (ref. 51) and OBFC1-locus SNP rs4387287 (ref. 52), which have similar effects on telomere length (75 bp and 115 bp per allele, respectively).
Our only findings consistent with the hypothesis that shorter telomeres predispose to increased cancer risk53 (equivalent to longer telomeres being protective) are those from the peak 1 SNPs. However, a regulatory element construct containing the longer telomere–associated alleles of three highly correlated SNPs, rs2736108, rs2736107 and rs2736109 (reconstructing a haplotype with 25% frequency in Europeans35), virtually abolished promoter activity in a reporter assay. This finding leaves a seemingly paradoxical association between lower enhancer activity and greater telomere length (Fig. 4). Control of telomerase activity is currently poorly understood, and this finding clearly merits further investigation.
SNPs within peak 3 (TERT introns 2–4) show strong associations with hormone-related cancers: peak 3 SNP rs10069690 is associated with risk of ER-negative breast cancer34 and breast cancer in BRCA1 mutation carriers, consistent with the observation that the majority of breast cancers arising in BRCA1 mutation carriers are ER negative. This variant has been reported to be associated with prostate cancer26, 54, and we find it associated with serous invasive ovarian cancer. Although SNPs in peaks 2 and 3 overlap on a physical map, the SNPs most strongly associated with cancer risk or telomere length were not highly correlated with each other (r2 between rs10069690 and rs7705526 = 0.33; Fig. 2b). This observation suggests that either the associations observed with multiple cancers and SNPs in peak 3 are mediated via a mechanism distinct from control of telomere length or that telomere length in breast, prostate and ovarian cells is under the control of a different set of SNPs from those controlling telomere length in leukocytes. Luciferase reporter assays show that peak 3 contains a silencer of the TERT promoter and that the minor allele of peak 3 SNP rs2242652 further reduces expression. Consistent with this finding, Kote-Jarai et al.54 report that the minor, risk allele of this SNP is associated with reduced TERT expression in benign prostate tissue. However, we were unable to identify comparable associations in ovarian or breast tumor tissue, possibly because TERT expression is severely dysregulated in most tumors. Taken together, our luciferase assays indicate that either reduced signal from regulatory elements in peaks 1 and 3 or increased signal from peak 2 increases risk of specific cancer types.
It should be noted that the minor allele of rs2242652 is associated with significantly lower risk of prostate cancer54 (OR = 0.84, 95% CI = 0.81–0.87; Ptrend = 1 × 10−23) but with significantly higher risks of breast and ovarian cancers (Tables 1 and 2). Similarly, a nearby SNP, rs401681, is associated with higher risks of cancers of the lung, bladder, testes and cervix and basal cell carcinoma but with lower risk of melanoma28, 30, 31. Such inverted associations might be due to tissue-specific interactions that need further examination.
We have additionally shown that the minor allele of rs10069690 affects splicing and is associated with transcription of a novel truncated isoform resulting from the introduction of a premature stop codon (Supplementary Fig. 6). We do not yet know whether this isoform affects canonical telomerase activity or how it changes activity. We further identified new open chromatin signatures overlapping rs10069690 in breast stromal and myoepithelial/stem cell fractions but not in progenitor or differentiated luminal epithelial cell fractions. Senescent stromal cells can stimulate malignant transformation of epithelial cells in in vitro and in vivo models55, 56, and the mechanisms mediated by these SNPs merit investigation in future studies.
The SNPs originally reported to be associated with risk of lung (rs402710)57 and breast (rs3816659)58 cancers (Supplementary Table 10) were not associated with any cancer in this study. Moreover, SNP rs2736100 in peak 2 has been reported to be associated with glioma and lung and testicular cancers27, 28, 31, 57, 59, 60, 61, 62, whereas nearby SNP rs2853677 was reported to be associated with glioma in the Han Chinese population63. Despite their physical proximity, these SNPs are not highly correlated with rs7705526 (r2 = 0.52 and 0.18, respectively), nor do they show independent associations with telomere length after adjustment for rs7705526. Thus, variants underlying susceptibility to different cancer types are different from the set of variants in the TERT region mediating changes in telomere length.
One limitation of this study is the incomplete representation of all SNPs at 5p15.33 on the iCOGS chip, which was designed in March 2010 using SNPs catalogued in HapMap 3 together with those from the pilot study of the 1000 Genomes Project35. To help fill known gaps on the iCOGS chip, additional SNPs were genotyped from the October 2010 1000 Genomes Project data release, and imputation was based on the most recent January 2012 release. However, several gaps remain across the TERT locus, and the existence of these gaps, coupled with the low linkage disequilibrium across the region (Fig. 2), raises the possibility that there could be more independent associations that we have not yet detected. Furthermore, the incomplete SNP catalog at the time of study design means that we cannot assume with certainty that the true causal variants, directly responsible for the observed association peaks, were captured in our analysis. It is also possible that additional rare variants not specifically investigated in this study could have functional effects within this locus. Further resequencing of this region is needed to uncover the full spectrum of variation and phenotype associations. Another limitation is that telomere length was measured in DNA from leukocytes rather than from breast or ovarian tissue. Whereas we obtained suitable blood DNA for measurements in >53,000 subjects (a necessary sample size for adequate statistical power), obtaining comparable qualities and quantities of DNA from normal breast or ovary cells would be almost impossible. Telomere lengths measured in different tissues within one individual have been shown to be highly correlated64, 65, 66, meaning that leukocyte telomere lengths are likely to be good surrogates for the corresponding lengths in other tissues. Furthermore, one of our aims was to investigate whether the previously reported associations between mean telomere length and cancer risk might be mediated by TERT variants, and such studies have been based on telomere length measured in blood cell DNA. Another limitation was that we were unable to stratify OCAC ovarian cancer cases by BRCA1 and BRCA2 mutation status because this information was not available; nor was there sufficient power to evaluate ovarian cancer risk in mutation carriers in CIMBA.
Our findings provide evidence relevant to the hypothesis that shorter telomeres increase cancer risks: associations in the TERT promoter (peak 1) fit this hypothesis best, whereas those in peaks 2 and 3 (TERT introns 2–4) and other reported 5p15.33 SNP cancer associations (Supplementary Table 10) do not. Thus, it would seem that the majority of cancer associations within the TERT locus are mediated via alternative mechanisms involving the TERT gene. The protein product of TERT has functions beyond the telomerase-mediated extension of telomeres67. These non-canonical functions of TERT strongly resemble those mediated by MYC and WNT68, which are upstream regulators of proliferation, differentiation and migration. TERT also modulates WNT/β-catenin signaling69, and ectopic TERT expression induces increased cell division and decreased apoptosis in primary mammary cells, independent of telomere elongation70.
In conclusion, this study provides definitive evidence for genetic control of telomere length by common genetic variants in the TERT locus. Additionally, we report multiple, independent TERT SNP associations with breast cancer risk, confirming previously reported associations and identifying new associations in both the general population and in BRCA1 mutation carriers. We also provide, for the first time to our knowledge, highly significant evidence for the association of distinct TERT SNPs with serous LMP and invasive ovarian cancer risks. Our results show that the relationships between TERT genotype, telomere length and cancer risk are complex and that the TERT locus may influence cancer risk through multiple mechanisms.
HapMap 3 catalog, http://www.sanger.ac.uk/resources/downloads/human/hapmap3.html; Wellcome Trust Case Control Consortium investigators, http://www.wtccc.org.uk/; investigators and institutions constituting the TCGA research network, http://cancergenome.nih.gov/.
SNP selection and genotyping.
Most SNPs were genotyped on the iCOGS custom array36, 37, 71. SNPs at 5p15.33 (Build 36 positions 1,280,000–1,415,000; Build 37 positions 1,227,693–1,361,969) were selected on the basis of published cancer associations, from the March 2010 release of the 1000 Genomes Project35. These included all known SNPs with MAF > 0.02 in Europeans and r2 > 0.1 with the then-known cancer-associated SNPs (rs402710 (ref. 57) and/or rs3816659 (ref. 58)), plus a tagging set for all known SNPs in the linkage disequilibrium blocks encompassing the genes in the region (SLC6A18, TERT and CLPTM1L). An additional 30 SNPs in TERT were selected through a telomere length candidate gene approach. In total, 134 SNPs were selected, 121 of which were successfully manufactured; 110 of those passed quality control36 in BCAC and CIMBA, and 108 passed quality control in OCAC (Supplementary Tables 1–3). After genotyping, these SNPs were complemented with 22 SNPs, selected from the October 2010 release of the 1000 Genomes Project to improve coverage. These were genotyped in two BCAC studies, SEARCH72 and CCHS73, using a Fluidigm array according to the manufacturer’s instructions. To improve SNP density further, comprehensive genotype data for the locus were imputed for all subjects on the basis of the January 2012 1000 Genomes Project release. The genotype imputation process is described in refs. 36,37,38. All participants provided written informed consent. Ethical approval for each study/consortium is described in detail in refs. 36,37,38.
Samples and quality control.
Study characteristics, iCOGS methodology and quality control for cancer risk analyses are detailed elsewhere36, 37, 38. We measured telomere length in 6,766 control samples from the SEARCH study; 1,569 of these were accrued by SEARCH itself36, 793 were collected as part of the Sisters in Breast Screening (SIBS) study15, and 4,404 were cancer-free participants in the European Prospective Investigation into Cancer (EPIC)-Norfolk study19. We also measured telomere length in 8,841 participants in CCHS73, 74 and in 38,145 participants in the Copenhagen General Population Study (CGPS)75, 76. Genotype clusters were visually inspected for the most strongly associated SNPs (Supplementary Fig. 2). For all studies, ancestry was assigned using HapMap (release 22) genotype data for European, African and Asian populations as reference (for BCAC and CIMBA, using multidimensional scaling; for OCAC, using LAMP77). All CIMBA analyses were restricted to individuals of European ancestry. For BCAC, separate estimates for individuals of east Asian and African-American ancestry were also derived. For OCAC, limited analyses of non-European ancestry groups were also performed. A subset of BCAC and OCAC cases and controls was used in previous breast and ovarian cancer association studies of individual SNPs78. However, the associations with the key SNPs (rs10069690, rs2736108 and rs7705526) remained significant after excluding this subset of cases and controls from analysis, demonstrating similar ORs.
Telomere length measurement.
Telomere length was measured in SEARCH using a modified version of the protocol described elsewhere19, 79. Twelve percent of samples were run in duplicate. Failed PCR reactions were not repeated. Telomere length was measured in CCHS and CGPS with a modified version of the protocol described elsewhere50, 80. Each individual was measured in quadruplicate. After exclusion of outliers, average cycle threshold (CT) values of the remaining samples were calculated. Failed measurements were repeated up to twice. For meta-analysis, telomere length measurements from SEARCH were converted to the same scale as that used for the CCHS and CGPS measurements on the basis of parameters from the linear regression between corresponding 5-percentile groups (including the 5th, 10th, 15th, 20th, 25th, 30th, 35th, 40th, 45th, 50th, 55th, 60th, 65th, 70th, 75th, 80th, 85th, 90th, 95th, 97th and 98th percentiles) in each 10-year age group of women from CCHS and SEARCH (Supplementary Fig. 7). This measure of telomere length was used for all the analyses and then converted into fold change (RTL) to aid interpretation (Supplementary Fig. 7).
SNP associations with telomere length were evaluated using linear regression to model the fold change in telomere length per minor allele, adjusted for age, 384-well plate, sex, 7 principal components and study. Each SNP was coded as the number of minor alleles (0, 1 or 2 for genotyped SNPs and the inferred genotype for imputed SNPs). The test of association was based on the 1-degree-of-freedom trend test statistic. We also performed separate analyses (SEARCH, CCHS females, CCHS males, CGPS females and CGPS males) and combined the parameter estimates in a fixed-effect meta-analysis in STATA (StataCorp). Associations with breast and ovarian cancer risks in BCAC and OCAC were evaluated by comparing genotype frequencies in cases and controls using unconditional logistic regression. Analyses were adjusted for study and by seven principal components in BCAC36 and five principal components in OCAC37. Nine OCAC studies with case-only genotype data were paired with case-control studies from similar geographic regions, resulting in 34 analysis study strata. The principal analysis fitted each SNP as an allelic dose and tested for association using a 1-degree-of-freedom trend test, but genotype-specific risks were also obtained. Associations between genotypes and breast cancer risk in CIMBA studies (BRCA1 mutation carriers) were evaluated using a 1-degree-of-freedom per-allele trend score test, based on modeling the retrospective likelihood of the observed genotypes conditional on breast cancer phenotypes81. To allow for non-independence among related individuals, an adjusted version of the score test was used in which the variance of the score was derived, taking into account the correlation between the genotypes by estimating the kinship coefficient for each pair of individuals using the available genotype data82. Per-allele HR estimates were obtained by maximizing the retrospective likelihood. All analyses were stratified by country of residence. US and Canadian strata were further stratified on the basis of reported Ashkenazi Jewish ancestry.
Conditional analyses were performed to identify SNPs independently associated with each phenotype. To identify the most parsimonious model, all SNPs with marginal P value < 0.001 were included in forward selection regression analyses with a threshold for inclusion of P < 1 × 10−4 and with terms for age (for telomere length only), principal components and study. Similarly, forward selection Cox regression analysis was performed for BRCA1 mutation carriers, stratified by country of residence, using the same P-value thresholds. This approach provides valid association tests, although the estimates can be biased81, 83. Parameter estimates for the most parsimonious model were obtained using the retrospective likelihood approach.
Normal breast tissue was donated by women undergoing reduction mammoplasty surgery. These individuals provided written consent, and all work was performed with full local institutional human ethics approval. Tissue was dissociated as described previously84. Cells were prepared for flow cytometry as described previously42 by staining with a cocktail of Lin+ markers (CD31-PE, CD45-PE and CD235a-PE), EpCAM-FITC, CD49f-PE-Cy5 and Sytox Blue. Cells were then processed by a BD FACSAria II Cell Sorter, and live cells negative for immunostaining of Lin+ markers were sorted into four subpopulations on the basis of their EpCAM-FITC and CD49f-PE-Cy5 fluorescence.
Cell pellets derived from FACS fractionation of breast tissue samples were cross-linked in 1% formaldehyde and lysed in 200 μl of Tris-buffered 1% SDS lysis buffer containing protease inhibitors. Lysates were sonicated using a QSONICA Model Q125 Ultra Sonic Processor to shear chromatin to fragments of 200 bp to 1 kb in length. Insoluble cell material was removed through centrifugation, and supernatants were equally divided into 100-μl input and FAIRE samples. Input samples were incubated overnight at 65 °C to reverse cross-linking. All samples were purified through two rounds of phenol-chloroform extraction, and DNA was recovered through ethanol precipitation and resuspended in water for use as PCR template. Sequences for PCR primers are listed in Supplementary Table 11.
Plasmid construction and luciferase assays.
TERT promoter variants were introduced into pGL3-TERT-3915 (ref. 43) by site-directed mutagenesis (Agilent Technologies). TERT PRE-A (hg19; chr. 5: 1,284,900–1,287,087) and PRE-B (chr. 5: 1,279,401–1,282,763) were PCR amplified using KAPAHiFi DNA polymerase (Geneworks) and cloned into pGL3-TERT-3915 or the vector encoding the minor alleles of rs2736107, rs2736108 and rs2736107. Individual SNPs were incorporated using overlap extension PCR. Sequences for PCR primers are listed in Supplementary Table 11. Cells were transfected with equimolar amounts of luciferase reporter plasmids and 50 ng of pRLTK using siPORT NeoFX Transfection Agent (Ambion), according to the manufacturer’s instructions, and harvested after 48 h. Luminescence activity was measured with a Wallac Victor3 1420 multilabel counter, and data from three replicates per construct were analyzed by one-way ANOVA with post-hoc Dunnett’s tests.
Mini-gene construction and quantitative RT-PCR analysis.
TERT intron 4 was synthesized by GenScript and subcloned into pIRES-TERT44. The minor alleles at rs10069690 and rs2242652 were introduced by site-directed mutagenesis (Agilent Technologies). The resultant plasmids, designated pIRES-TERTint4-WT (wild-type intron 4), pIRES-TERTint4-rs10069690, pIRES-TERTint4-rs2242652 and pIRES-TERTint4-DM (minor alleles at both sites), were transfected into cells using siPORT NeoFX Transfection Agent, and cells were harvested after 24 h. Total RNA was extracted using the RNeasy Mini kit (Qiagen) and digested with DNase I (Invitrogen). cDNA was synthesized from 1 μg of RNA by random priming using SuperScript III reverse transcriptase (Invitrogen). Samples were screened for the presence of TERT splice variants by RT-PCR. Sequences for PCR primers are listed in Supplementary Table 11.
Molecular correlations at the 5p15.33 locus.
For each gene within 1 Mb of the TERT locus, we performed the following assays: (i) gene expression analysis in ovarian cancer cell lines (n = 50) compared to ovarian surface epithelial and fallopian tube secretory cell lines (n = 73) and tissues from high-grade serous ovarian cancers; (ii) methylation analysis in high-grade serous ovarian cancers compared to normal tissues and methylation quantitative trait locus (mQTL) analysis; and (iii) expression quantitative trait locus (eQTL) analysis to evaluate genotype–gene expression associations in normal high-grade serous ovarian cancer precursor tissues. We also evaluated these genes in silico in the somatic data from TCGA49. We profiled the spectrum of noncoding regulatory elements in ovarian surface epithelial and fallopian tube secretory cell lines using a combination of FAIRE sequencing (FAIRE-seq40) and RNA sequencing (RNA-seq).
The U.S Dollar is quickly losing its status as the world reserve currency. Five of the top ten economies in the world, plus a few others, no longer use the dollar as an intermediary currency for trade. This trend poses a huge risk to the dollar and the United States along with it.
ZeroHedge points out today that Australia, the world’s 12th-ranked economy, has now joined a growing list of nations that have agreed to bypass the dollar in bilateral trade with China. China, ranked 2nd behind the U.S., also has similar agreements with Japan (3rd), Brazil (6th), India (9th), and Russia (10th).
Although unilateral agreements have been in place for some time between China and the countries listed above, last week the BRICS (Brazil, Russia, India, China and South Africa) agreed to set up a development bank to compete with the IMF, indicating it’s gearing up to compete in a post-dollar world.
Additionally, Brazil, who agreed in principle to drop the dollar with bilateral trade with China some time ago, just made it official with $30 billion in annual currency swaps which will facilitate around 50% of all trade between them.
Besides those agreements with China, some of these nations have made other similar agreements with each other. India and Japan began swapping $15 billion in each other’s currency in 2011 to handle their bilateral trade. And the sanctions against Iran haven’t stopped them from trading oil with China, Russia, and India in anything but the dollar.
It appears that the dollar is certainly nearing the end of its reign, which could lead to severe economic hardship for the United States.
Dave Hodges writes:
The United States’ good economic fortune is due solely to the fact that world must use the dollar, the Petrodollar if you will, in order to make their nation’s individual oil purchases; this provides the only source of backing for the U.S. dollar that the Federal Reserve requires in order to somewhat sustain our back-breaking debt that the banker-occupied United States government has passed along to the American taxpayer in the form of bailouts.
And Marin Katusa of Casey Research writes:
If the US dollar loses its position as the global reserve currency, the consequences for America are dire. A major portion of the dollar’s valuation stems from its lock on the oil industry – if that monopoly fades, so too will the value of the dollar. Such a major transition in global fiat currency relationships will bode well for some currencies and not so well for others, and the outcomes will be challenging to predict. But there is one outcome that we foresee with certainty: Gold will rise. Uncertainty around paper money always bodes well for gold, and these are uncertain days indeed.
America’s imperialism, combined with its ultra-fiat status of unending debt creation, appears to have created a final downward spiral that has caused many of the top economies to abandon a sinking ship. It might not be too much longer before the rest follow suit. Now might be a great time to consider diversifying into other currencies, and even digital currencies, to mitigate growing losses in the U.S. dollar.
The U.S. economy is in a bubble inflated by “phony money” from the Federal Reserve and will burst within a few years, warned David Stockman, who was budget director for President Ronald Reagan.
In an essay published in the New York Times, Stockman wrote that the Fed’s quantitative easing policies in the aftermath of the credit crisis have flooded stock markets with cash even while the “Main Street economy” remains weak. The combination, he wrote, is “unsustainable.”
“When it bursts, there will be no new round of bailouts like the ones the banks got in 2008,” wrote Stockman, a former senior managing director at Blackstone Group LP and a former Republican congressman from Michigan.
“Instead, America will descend into an era of zero-sum austerity and virulent political conflict, extinguishing even today’s feeble remnants of economic growth.”
Stockman, 66, is the author of “The Great Deformation: The Corruption of Capitalism in America,” which will be published April 2.
The Fed, led by Ben S. Bernanke, is purchasing $85 billion in assets every month. The Fed is leaving its key interest rate near zero while it tries to reduce unemployment below 6.5 percent and hold inflation below 2.5 percent.
The Standard & Poor’s 500 Index rose to an all-time high last week, closing at 1,569.19 on March 28. That surpassed the previous record of 1,565.15 set in October 2007. U.S. stock markets were closed March 29 for the Good Friday holiday.
Among the other culprits Stockman blamed for what he termed a “state-wreck” are President Franklin Delano Roosevelt for weakening the gold standard in 1933, President Richard Nixon for removing the convertibility of dollars to gold and “lapsed hero” Alan Greenspan, the former Fed chairman, for keeping interest rates too low for too long.
Investors will sell, Stockman wrote, at any hint that the Fed is starting to remove assets from its balance sheet.
“Notwithstanding Bernanke’s assurances about eventually, gradually making a smooth exit, the Fed is domiciled in a monetary prison of its own making,” he wrote, warning of unsustainable fiscal policies as well. “These policies have brought America to an end-stage metastasis. The way out would be so radical it can’t happen.”
Paul Krugman, the Princeton University economist and New York Times columnist, responded on his blog yesterday, saying that he was “disappointed” in Stockman’s “gee-whiz, context- and model-free numbers embedded in a rant — and not even an interesting rant.”
Krugman called Stockman’s piece “cranky old man stuff,” and summarized it this way:
“We’ve been doomed, yes doomed, ever since FDR took us off the gold standard and introduced unemployment insurance. What about those 80 years of non-doom? Just a series of lucky accidents. Now we’re really doomed. I mean it!”
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Former Florida Gov. Jeb Bush has written the cover story for the April edition of Newsmax magazine, offering an optimistic view of America’s future — and straight talk about its problems — with the resounding message: “Growth is the answer” to our nagging economic woes.
And the leading Republican sat down with Newsmax for a wide-ranging interview elaborating on a number of topics he discusses in his magazine feature, arguing that America can be great again if we open up the doors to economic opportunity.
Bush, who served two terms as Florida’s governor, has earned a nationwide reputation for reform proposals in the fields of education and immigration. He is founder and chairman of the Foundation for Excellence in Education, author of the new book “The Immigration Wars: Forging an American Solution” — co-written with constitutional law expert Clint Bolick — and is considered a strong contender for the GOP presidential nomination in 2016.
Bush’s Newsmax magazine piece stresses the need for economic growth to deal with the nation’s debt and deficit problems.
In his exclusive interview with Newsmax TV, he was asked if growth alone will solve those problems.
Story continues below.
“If we grew at 2 percent more per year than what’s projected, which is anywhere from 1 to 1.5 percent per year in real terms, we would create a Germany in terms of incremental economic output in the 10th year,” he says. “And that Germany would be taxed at the current rate at least to the tune of about $1 trillion.
“There is no tax plan that could come close to [that].
“The president, almost every time he has a chance to speak, talks about the need for high growth, but his policies do the exact opposite. His policies have created the most tepid recovery in modern American history.
“The dual effect of high growth creating higher income that’s taxed by government at all levels, combined with lessening demands placed on government that occurs during economic prosperity, is a worthy objective.
“Right now we have this discussion of austerity versus government spending, when in fact what we ought to be doing is [determining] how we can invest in the private sector, creating investment opportunities and job creating opportunities that create the kind of growth that lessens government’s involvement in all this. That’s what’s missing and the president has a duty to outline specific proposals that will create economic growth. Up until now I don’t think he’s done it.”
Bush suggests that Republicans could be talking too much about the debt and deficit instead of growth.
“It’s a principled position to say that we have to live within our means, and over time you can do that and we should do that. But the fastest way to get to that is in combination with high growth. It’s also a winning political message to embrace the dynamic nature of our economy. It’s much better than just saying, ‘world coming to end. Memo to follow.’
“We should be much more hopeful and optimistic about the future of our country. If you just see the trends in the life science area and telecommunications and information technology, we’re living in the most wondrous time ever in human history, and globally the United States is a leader. We should be excited about the future and creating strategies to ensure that we create long-term, sustained economic growth.”
Reform should start with energy, immigration policy, regulations, the tax code, and education, Bush states.
“They’re the set of policy objectives that we should embrace, not all of which are ideological either. The sad thing is the dysfunction in Washington right now limits the ability even when there’s enough consensus to move forward.”
During the presidential campaign, Mitt Romney was criticized for not putting specifics on the table, particularly regarding entitlement cuts and taxes. But Bush believes lawmakers and candidates can get specific without getting bogged down in minutiae.
“First of all you have to inform. You can’t just assume that everybody has the facts,” he maintains.
“Most of the cuts we’re talking about are cuts in the growth of spending, not actual cuts, and there are people in Washington that have [offered] specific proposals, Paul Ryan being perhaps the most obvious one. He was Mitt Romney’s running mate and a courageous and inspired choice, and they did defend those so-called cuts for a while, then they kind of reverted back to trying to make the election about the president, exclusively a referendum on his job performance.
“. . . The lesson of 2012 is be bold, be specific, persuade, don’t follow polls, don’t be driven by what people’s feelings are here and now because those opinions can shift if you make a compelling case.”
But Bush does agree that the entitlement system is “unsustainable” and “no one argues that we can keep doing what we’re doing. So either we have it collapse or we change it to protect it.”
Among the changes he suggests are “raising the retirement age to reflect the life expectancy increase that’s been dramatic, means testing some of the entitlement programs over time. We have to reform healthcare underneath the entitlement system as well so that the cost curve is dealt with, which means we should move toward catastrophic coverage as the form of insurance and reward healthy lifestyle decisions and focus on prevention to lessen cost by improving healthcare outcomes.”
Bush favors eliminating tax loopholes and reducing taxes to the lowest possible rates
“If you eliminate as many of the deductions as possible, you would create a boom of economic activity. You would lessen compliance costs. Right now, whether it’s a small business guy or a private person, people pay an arm and a leg for just the simple compliance costs. Now, you have to pay a lot of money because our code’s way too complex. My personal belief is the simpler the code, the better it is.”
Asked if he would be willing to give up the mortgage interest deduction if tax rates are lowered, Bush responds: “I would. To me, the better way of doing this would be to say, all of the tax code expires” and must be rebuilt from the ground up.
“That would be a better approach because people could see the benefits of a simpler code and the empowering nature of that. People could keep more of their own money, make decisions for themselves, and not effectively get in line to get their version of some tax expenditure or tax credit. [We would] lower the rates significantly and create a burst of economic activity for our country.”
In his Newsmax magazine article, Bush never once mentions President Obama or the Democrats.
He tells Newsmax TV that other Republicans could be too focused on talking about what the opposition is doing instead of what they have to offer.
“We’ve gotten a little far removed from where we once were not that long ago, which is the party of reform, the party of new thinking, the party of ideas, the party that embraces the future.
“There’s a reason why in my recent speeches and the Newsmax article that I didn’t mention the president. The president’s record on economic policy is a failed one, but we’re not going to win the day unless we have a compelling alternative and that’s where the focus needs to be.
“In Washington, particularly, the loyal opposition has a job to be the opposition. But you can’t stop there. You have to oppose in a principled, civil way but you also have to offer alternatives.
“There are a lot of reasons why the election turned out the way it did. All I know for sure is that I wish Mitt Romney was president right now because he’d be a practical person trying to forge common-sense solutions to these big, dysfunctional challenges we face.”
On education, Bush writes in his magazine cover story about the need to increase the choices that are available to parents and students.
To do that, he tells Newsmax we need to “change laws. We have 50 states that are the governance model or the catalyst for how governance operates. We have 13,000 government-run monopolies, heavily unionized. And in that kind of setting, it should not be a surprise that the economic interests of the adults is really a dominant part of education life in America today.
“We have a third of our kids that don’t make it through the system, even though we spend more per student than any country in the world. And a lot of students could be doing college-level work by the time they’ve graduated from high school but in effect they’re held back because we’ve got this adult-centered homogenized learning model.
“What the Foundation for Excellence in Education is trying to do is to help policymakers in our 50 states open up their laws to allow more innovation to take place.
“We’re putting too much emphasis on the college experience. The country’s moving towards fewer standards that require critical thinking skills both for going to college as well as for being able to get a job in some technical skill or career-oriented avocation. So the skills necessary for both paths need to be higher.”
Bush in his magazine article cites the need to strengthen family values as a way to boost our economy.
“It’s really important to recognize that family life in America has changed pretty dramatically,” he tells Newsmax TV.
“For the first time in American history there are fewer married women than unmarried women in the age cohort that’s measured. We’ve had dramatic increases in out-of-wedlock birth rates. Every study that I’ve seen suggests that if a young woman doesn’t have a child before she’s married and she can defer marriage [until] she’s 25 and can graduate from high school, the chance of her living in poverty is dramatically reduced, and similarly with men.
“If we get that right then we can have a chance of creating economy prosperity over the long haul. If we don’t get it right, it really does change who we are as a nation and it creates enormous strains.“
Bush also says there is an “inverse relationship” between liberty and the size and scope of government, and agrees with Sen. Rand Paul that the Republican Party needs to embrace more libertarian ideas.
He also talks at length about the immigration policies espoused in his controversial new book.
And he ends his Newsmax interview on an optimistic note, stating that “I just have total confidence that a dynamic world will yield benefits that we can’t even imagine.
“If you believe like I do that the world is abundant with possibilities, then we need to make sure we build capacity so that everybody is successful or can be successful in the pursuit of their dreams — not the dreams of someone from government, but their own dreams.”
People often boggle at the use of the word “neoliberal”; as if the utterer were some kind of crazed tinfoil hat wearing conspiracy theorist raving about insane lizard-man conspiracies, rather than someone attempting to concisely define the global economic orthodoxy of the last three decades or so.
One of the main problems we encounter when discussing neoliberalism is the haziness of the definition. Neoliberalism is certainly a form of free-market neoclassical economic theory, but it quite difficult to pin down further than that, especially since neoliberal governments and economists carefully avoid referring to themselves as neoliberals and the mainstream media seem to avoid using the word at all costs (think about the last time you saw a BBC or CNN news reporter use the word “neoliberal” to describe the IMF or a particularly right-wing government policy).
The economic model that the word “neoliberalism” was coined to describe was developed by the “Chicago school” economists in the 1960s and 1970s based upon Austrian neoclassical economic theories, but heavily influenced by Ayn Rand’s barmy pseudo-philosophy of Übermenschen and greed-worship.
The first experiment in applied neoliberal theory began on September 11th 1973 in Chile when a US backed military coup resulted in the death of social-democratic leader Salvador Allende and his replacement with the brutal military dictator General Pinochet. Thousands of people were murdered by the Pinochet regime for political reasons and tens of thousands more were tortured, as Pinochet and the “Chicago boys” set about implementing neoliberal economic reforms and brutally suppressing anyone that stood in their way. The US financially doped the Chilean economy in order to create the impression that these rabid-right wing reforms were successful. After the “success” of the Chilean neoliberal experiment the instillation and economic support of right-wing military dictatorships to impose neoliberal economic reforms became unofficial US foreign policy.
The first of the democratically elected neoliberals were Margaret Thatcher in the UK and Ronald Reagan in the US. They both set about introducing ideologically driven neoliberal reforms, such as the complete withdrawal of capital controls by Tory Chancellor Geoffrey Howe and the deregulation of the US financial markets that led to vast corruption scandals like Enron and the neoliberal economic meltdown. By 1989 the ideology of neoliberalism was enshrined as the economic orthodoxy of the world as undemocratic Washington based institutions such as the International Monetary Fund (IMF), the World Bank and the US Treasury Department signed up to a ten point economic plan which was riddled with neoliberal ideology such as trade liberalisation, privatisation, financial sector deregulation and tax cuts for the wealthy. This agreement between anti-democratic organisations is misleadingly referred to as “The Washington Consensus”.
These days the IMF is the most high profile pusher of neoliberal economic policies. Their strategy involves applying strict “structural adjustment” conditions on their loans. These conditions are invariably neoliberal reforms such as privatisation of utilities and government owned industries, tax cuts for corporations and the wealthy, the abandonment of capital controls, the removal of democratic controls over central banks and monetary policy and the deregulation of financial industries.Neoliberal economic policies have created economic disaster after economic disaster, virtually wherever they have been tried out. Some of the most high profile examples include:
South Africa: When the racist Apartheid system was finally overthrown in 1994, the new ANC government embraced neoliberal economic theory and set about privatising virtually everything, cutting taxes for the wealthy, destroying capital controls and deregulating their financial sector. After 18 years of neoliberal government, more black South Africans are living in extreme poverty, more people are unemployed and South Africa is an even more unequal society than it was under the racist Apartheid regime. Between 1994 and 2006 the number of South Africans living on less than $1 a day doubled from 2 million to 4 million, by 2002, eight years after the end of Apartheid 2002 the unemployment rate for black South Africans had risen to 48%.*
Russia: After the fall of communism, neoliberal economists flooded into Russia to create their free-market utopia, however all they managed to do was massively increase levels of absolute poverty, reduce productivity and create a few dozen absurdly wealthy oligarchs who siphoned their $trillions out of Russia to “invest” in vanity projects such as Chelsea FC. Within less than a decade of being one of the world’s two great super-powers, the neoliberal revolution resulted in Russia defaulting on their debts in 1998.
Argentina: Praised as the poster-boys of neoliberalism by the IMF in the 1990s for the speed and scale of their neoliberal reforms, the Argentine economy collapsed into chaos between 1999-2002, only recovering after Argentina defaulted on their debts and prioritised repayment of their IMF loans, which allowed them to tear up the IMF book of neoliberal dogma and begin implementing an investment based growth strategy which boosted the Argentine economy out of their prolonged recession. The late Argentine President Néstor Kirchner famously stated that the IMF had “transformed itself from being a lender for development to a creditor demanding privileges“.
The Eurozone: The right-wing love to drivel on about how the EU is a “leftie” organisation, but the unelected technocrats that run the EU (the European commission and the European Central Bank) are fully signed up to the neoliberal economic orthodoxy, where economic interests are separated from democratic control. Take the economic crisis in Greece: The EC and the ECB lined up with the neoliberal pushing IMF to force hard line neoliberal reforms onto the Greek economy in return for vast multi-billion “bailouts” that flowed directly out of Greece to “bail out” their reckless creditors (mainly German and French banks). When the neoliberalisation reforms resulted in further economic contraction, rising unemployment and worsening economic conditions the ECB, EC, IMF troika simply removed the democratic Greek government and appointed their own stooge, an economic coup trick they also carried out in Italy. Spain and Ireland are other cracking examples of neoliberal failure in the Eurozone. These two nations were more fiscally responsible than Germany, France or the UK in terms of government borrowing before the neoliberal economic meltdown, however their deregulated financial sectors inflated absurd property bubbles, leaving the Irish and Spanish economies in ruins once the bubbles burst around 2007-08.
Despite this litany of economic failures, neoliberalism remains the global economic orthodoxy. Just like any good pseudo-scientific or religious orthodoxy the supporters of neoliberal theory always manage to come up with a load of post-hoc rationalisations for the failure of their theories and the solutions they present for the crises their own theories induced are always based upon the implementation of even more fundamentalist neoliberal policies.
One of the most transparent of these neoliberal justification narratives is the one that I describe as the Great Neoliberal Lie: The fallacious and utterly misleading argument that the global economic crisis (credit crunch) was caused by excessive state spending, rather than by the reckless gambling of the deregulated, neoliberalised financial sector.
Just as with other pseudo-scientific theories and fundamentalist ideologies, the excuse that “we just weren’t fundamentalist enough” is always there. The neoliberal pushers know that pure free-market economies are as much of an absurd fairytale as 100% pure communist economies, however they keep pushing for further privatisations, tax cuts and deregulations precicely because they are the direct beneficiaries of these policies. Take the constantly widening wealth gap in the UK throughout three decades of neoliberal policy. The minority of beneficiaries from this ever widening wealth gap are the business classes, financial sector workers, the mainstream media elite and the political classes. It is no wonder at all that these people think neoliberalism is a successful ideology. Within their bubbles of wealth and privilege it has been, to everyone else it has been an absolute disaster.
Returning to a point I raised earlier in the article; one of the main problems with the concept of “neoliberalism” is the nebulousness of the definition. It is like a form of libertarianism, however it completely neglects the fundamental libertarian idea of non-aggression, in fact it is so closely related to that other (highly aggressive) US born political ideology of Neo-Conservatism that many people get the two concepts muddled up. A true libertarian would never approve of vast taxpayer funded military budgets and the waging of imperialist wars of aggression.
Another concept that is closely related to neoliberalism is the ideology of minarchism (small stateism), however the neoliberal brigade seem perfectly happy to ignore the small-state ideology when it suits their personal interests. Take the vast banker bailouts (the biggest state interventions in human history) that were needed to save the neoliberalised global financial sector from the consequences of their own reckless gambling or the Tory policy of bringing in government imposed alcohol price fixing measures.
The Godfather of neoliberalism was Milton Friedman. He made the case that illegal drugs should be legalised in order to create a free-market drug trade, this is politically inconvenient, so unlike so many of his neoliberal ideas that have consistently failed yet remain incredibly popular with the wealthy elite, Friedman’s drug legalisation proposals have never even been tried out.
The fact that neoliberals are so often prepared to ignore the fundamental principles of libertarianism (the non-aggression principle, drug legalisation) and abuse the fundamental principles of small state minarchism (vast taxpayer funded bailouts for their financial sector friends, £billions in taxpayer funded outsourcing contracts, alcohol price fixing schemes) demonstrate that neoliberalism is actually more like Ayn Rand’s barmy (greed is the only virtue, all other “virtues” are aberrations) pseudo-philosophical ideology of objectivism than a set of formal economic theories.
Neoliberal fanatics in powerful positions have demonstrated time and again that they will willingly ditch their right-wing libertarian and minarchist “principles” if those principles happen to conflict with their own personal self-interest. Neoliberalism is less of a formal set of economic theories than an error strewn obfuscation narrative to promote the economic interests, and justify the personal greed of the wealthy, self-serving establishment elite.
There has been an ongoing battle between researchers and the natural gas and oil industries over whether or not hydraulic fracturing (fracking) is definitively leading to an increase in earthquake activity.
Since September of 2010, nearly 1,000 earthquakes have rattled Arkansas and the area around the New Madrid Fault Line. Previous to this, Arkansas had a total of 38 quakes in 2009. Yet two cities, Greenbrier and Guy had a swarm of 30 small earthquakes in a four-day period in early 2011, which paralleled fracking activity in the same area.
Now, a study has appeared from The Geology Society of America, which investigated the largest of the quakes that rattled Oklahoma and 17 other states in November of 2011. While supporting the research of others in establishing a causal link between fracking and earthquakes, they appear to have found another even more troubling aspect to the data.
According to the RMA, “The Rocky Mountain Arsenal deep injection well was constructed in 1961, and was drilled to a depth of 12,045 feet” and 165 million gallons of Basin F liquid waste, consisting of “very salty water that includes some metals, chlorides, wastewater and toxic organics” was injected into the well during 1962-1966.
Why was the process halted? “The Army discontinued use of the well in February 1966 because of the possibility that the fluid injection was “triggering earthquakes in the area,” according to the RMA. (Source)
They drew this conclusion even after the EPA stated that this method of deep injection was safe. Over the decades that would follow, both the U.S. Army Corps of Engineers and the U.S. Geological Survey would accumulate data that showed a link between fracking and underground instability that can trigger earthquakes.
One of the first large studies of an Oklahoma earthquake swarm which began early in 2011 was conducted by the Oklahoma Geological Survey. The results were released in August of 2011 and conclude with a probable correlation within this event, as well as citing a connection made from the historical record:
The strong correlation in time and space as well as a reasonable fit to a physical model suggest that there is a possibility these earthquakes were induced by hydraulic fracturing.
Our analysis showed that shortly after hydraulic fracturing began small earthquakes started occurring, and more than 50 were identified, of which 43 were large enough to be located. Most of these occurred within a 24 hour period after hydraulic fracturing operations had ceased. There have been previous cases where seismologists have suggested a link between hydraulic fracturing and earthquakes, but data was limited, so drawing a definitive conclusion was not possible for these cases. The first case occurred in June 1978 in Carter and Love Counties, just south of Garvin County, with 70 cases in 6.2 hours. The second case occurred in Love County with 90 earthquakes following the first and second hydraulic fracturing stages. [Nicholson and Wesson, 1990] (Source)
Given the aforementioned documentation by the U.S. Army and the U.S. Geological Survey, the data is beginning to look conclusive.
However, there is one more concern that is addressed in the most recent study conducted by the Geological Society of America, a non-profit organization with tens of thousands of members from nearly 100 countries. According to the abstract of this study, which looked most specifically at the largest earthquake reported, a 5.7 event that occurred in November (after the study cited above):
Significant earthquakes are increasingly occurring within the continental interior of the United States, including five of moment magnitude (Mw) ≥ 5.0 in 2011 alone. Concurrently, the volume of fluid injected into the subsurface related to the production of unconventional resources continues to rise. Here we identify the largest earthquake potentially related to injection, an Mw 5.7 earthquake in November 2011 in Oklahoma. The earthquake was felt in at least 17 states and caused damage in the epicentral region. It occurred in a sequence, with 2 earthquakes of Mw 5.0 and a prolific sequence of aftershocks.
Subsurface data indicate that fluid was injected into effectively sealed compartments, and we interpret that a net fluid volume increase after 18 yr of injection lowered effective stress on reservoir-bounding faults. Significantly, this case indicates that decades-long lags between the commencement of fluid injection and the onset of induced earthquakes are possible, and modifies our common criteria for fluid-induced events. The progressive rupture of three fault planes in this sequence suggests that stress changes from the initial rupture triggered the successive earthquakes, including one larger than the first [emphasis added]. (Source)
This is a worrisome conclusion which seems to indicate that even if fracking is halted, earthquake activity can continue as a result of previous activity, and earthquake magnitude can increase over time.
Much more studying needs to be done to confirm the above, and the full results still need to be scrutinized by the scientific community. However, in light of the worries already surrounding the historically dangerous New Madrid Fault Line, we should hope that definitive action is taken sooner rather than later to halt a process that is dubious at best, and cataclysmic at worst.
Note: beyond the risk of earthquakes, fracking has other horrendous consequences. To see what life is like for some residents within fracking zones, please do not miss the film Gasland.
WASHINGTON (The Borowitz Report)—Justice Antonin Scalia dropped a bombshell on the Supreme Court today, announcing his decision to resign from the Court “effective immediately” and leave the United States forever.
Calling this week “by far the worst week of my life,” Justice Scalia lashed out at his fellow-Justices and the nation, saying, “I don’t want to live in a sick, sick country that thinks the way this country apparently thinks.”
Justice Scalia said that he had considered fleeing to Canada, “but they not only have gay marriage but also national health care, which is almost as evil.”
He said the fact that nations around the world recognizing same-sex marriage are “falling like deviant dominoes” would not deter him from leaving the United States: “There are plenty of other countries that still feel the way I do. I’ll move to Iran if I have to.”
Throwing off his robe in a dramatic gesture, Justice Scalia reserved his harshest parting shot for his fellow-Justices, screaming, “Damn you! Damn each and every one of you to hell! You call yourself judges? That’s a good one. You’re nothing but animals!”
Breathing heavily after his tirade, he turned to Justice Clarence Thomas and said, “Except you, Clarence. Are you coming with me?”
Justice Thomas said nothing in reply.