Social and cultural influences on genetic screening programme acceptability: A mixed-methods study of the views of adults, carriers, and family members living with thalassemia in the UK.

As population-level carrier screening panels for reprogenetic information emerge globally, conditions to be included, and the timing of implementation is widely debated. Thalassemia is the only condition for which population-based prenatal carrier screening is offered in the UK. However, little is known about the views and experiences of the UK thalassemia-affected community toward this screening or other forms of genetic screening for thalassemia (newborn, preconception), despite the range of direct consequences of screening programmes for this group. Using a mixed-methods integrative analysis (qualitative interviews n = 20 and quantitative survey n = 80), this study outlines the experiences and attitudes of adults with thalassemia, their family members, and screen-identified thalassemia carriers toward preconception, prenatal, and newborn screening for thalassemia. The majority of participants described thalassemia as a burdensome condition with a range of negative impacts, which contributed to their strong support for screening in all its potential formats. However, the data also highlight the challenges of each screening mode for this group, reflected in the high level of value conflict in participants' accounts and decisions. Cultural, social, and (to a lesser extent) religious factors were found to mitigate against the advantages of early screens, particularly within faith communities. Social stigma emerged as key to this process, informing the way that thalassemia severity was not only perceived, but also experienced by affected adults, which ultimately influenced screening uptake and outcomes. These findings suggest that cultural and social sensitivity is as important as the mode of screening delivery itself, if the iatrogenic and unintended harms of screening-particularly the social/psychological burden of value conflict-are to be adequately addressed and minimized.

Newborn screening for haemophilia: The views of families and adults living with haemophilia in the UK.

INTRODUCTION: As genomic sequencing become more efficient and cost-effective, the number of conditions identified through newborn screening globally is set to dramatically increase. Haemophilia is a candidate condition; however, very little is known about the attitudes of the haemophilia community towards screening. AIM: This study aimed to outline the perspectives of adults with haemophilia and their families towards newborn screening. METHODS: A paper and online survey on screening were distributed to every family known to the Haemophilia Society UK. Data collection occurred between January and June 2018. In total, 327 participants completed the survey: 76% were a relative of a person with haemophilia and 24% had haemophilia themselves; 83% were living with haemophilia A and 17% with haemophilia B. RESULTS: The vast majority supported newborn screening (77%) and preferred it to other forms of screening (preconception or prenatal). Participants supported newborn screening primarily because they viewed it as a means to facilitate early support and treatment, facilitate informed decisions about future pregnancies and prevent the "diagnostic odyssey." The 23% who did not support the screen did not associate these particular benefits with newborn screening. CONCLUSION: Haemophilia emerged from this analysis as a condition that the vast majority of participants considered a "liveable" disability and one best suited to newborn screening programmes that could improve support to affected families rather than reduce the birth rate of affected children.

The role of experiential knowledge within attitudes towards genetic carrier screening: A comparison of people with and without experience of spinal muscular atrophy.

PURPOSE: Autosomal recessive conditions, while individually rare, are a significant health burden with limited treatment options. Population carrier screening has been suggested as a means of tackling them. Little is known, however, about the attitudes of the general public towards such carrier screening and still less about the views of people living with candidate genetic diseases. Here, we focus on the role that such experience has on screening attitudes by comparing views towards screening of people with and without prior experience of the monogenetic disorder, Spinal Muscular Atrophy. METHODS: An exploratory sequential mixed methods design was adopted. In-depth qualitative interviews were used to develop two surveys. The surveys addressed attitudes towards carrier screening (pre-conceptual and prenatal) for SMA. PARTICIPANTS: 337 participants with SMA experience completed the SMA Screening Survey (UK) and 336 participants with no prior experience of SMA completed the UK GenPop Survey, an amended version of the SMA Screening Survey (UK). RESULTS: The majority of both cohorts were in favour of pre-conception and prenatal carrier screening, however people with experience of type II SMA were least likely to support either. Key differences emerged around perceptions of SMA, with those without SMA experience taking a dimmer view of the condition than those with. CONCLUSION: This study underscores the significance of prior experience with the condition to screening attitudes. It highlights the need for accurate and high-quality educational resources to support any future carrier screening programmes, that particularly in relation to rare genetic disorders like SMA that will fall outside the remit of everyday experience for the majority of the population.

Impairment Experiences, Identity and Attitudes Towards Genetic Screening: the Views of People with Spinal Muscular Atrophy.

Developments in genetics are rapidly changing the capacity and scope of screening practices. However, people with genetic conditions have been under-represented in the literature exploring their implications. This mixed methods study explores the attitudes of people with Spinal Muscular Atrophy (SMA) towards three different population-level genetic screening programmes for SMA: pre-conception, prenatal and newborn screening. Drawing on qualitative interviews (n = 15) and a survey (n = 82), this study demonstrates that more severely affected individuals with early-onset symptoms (Type II SMA), are less likely to support screening and more likely to view SMA positively than those with milder, later onset and/or fluctuating symptoms (Types III/ IV SMA). Indeed, this clinically milder group were more likely to support all forms of screening and view SMA negatively. This paper highlights that screening is a complex issue for people with genetic conditions, and the nature of impairment experiences plays a critical role in shaping attitudes.

The SMN structure reveals its crucial role in snRNP assembly.

The spliceosome plays a fundamental role in RNA metabolism by facilitating pre-RNA splicing. To understand how this essential complex is formed, we have used protein crystallography to determine the first complete structures of the key assembler protein, SMN, and the truncated isoform, SMNΔ7, which is found in patients with the disease spinal muscular atrophy (SMA). Comparison of the structures of SMN and SMNΔ7 shows many similar features, including the presence of two Tudor domains, but significant differences are observed in the C-terminal domain, including 12 additional amino acid residues encoded by exon 7 in SMN compared with SMNΔ7. Mapping of missense point mutations found in some SMA patients reveals clustering around three spatial locations, with the largest cluster found in the C-terminal domain. We propose a structural model of SMN binding with the Gemin2 protein and a heptameric Sm ring, revealing a critical assembly role of the residues 260-294, with the differences at the C-terminus of SMNΔ7 compared with SMN likely leading to loss of small nuclear ribonucleoprotein (snRNP) assembly. The SMN complex is proposed to form a dimer driven by formation of a glycine zipper involving α helix formed by amino acid residues 263-294. These results explain how structural changes of SMN give rise to loss of SMN-mediated snRNP assembly and support the hypothesis that this loss results in atrophy of neurons in SMA.

Newborn screening for spinal muscular atrophy: The views of affected families and adults.

Spinal muscular atrophy (SMA) is one of the leading genetic causes of infant death worldwide. However, due to a lack of treatments, SMA has historically fallen short of Wilson-Jungner criteria. While studies have explored the acceptability of expanded newborn screening to the general public, the views of affected families have been largely overlooked. This is in spite of the potential for direct impacts on them and their unique positioning to consider the value of early diagnosis. We have previously reported data on attitudes toward pre-conception and prenatal genetic screening for SMA among affected families (adults with SMA [n = 82] and family members [n = 255]). Here, using qualitative interview [n = 36] and survey data [n = 337], we report the views of this same cohort toward newborn screening. The majority (70%) of participants were in favor, however, all subgroups (except adults with type II) preferred pre-conception and/or prenatal screening to newborn screening. Key reasons for newborn screening support were: (1) the potential for improved support; (2) the possibility of enrolling pre-symptomatic children on clinical trials. Key reasons for non-support were: (1) concerns about impact on the early experiences of the family; (2) inability to treat. Importantly, participants did not view the potential for inaccurate typing as a significant obstacle to the launch of a population-wide screening program. This study underscores the need to include families affected by genetic diseases within consultations on screening. This is particularly important for conditions such as SMA which challenge traditional screening criteria, and for which new therapeutics are emerging.

Population screening for spinal muscular atrophy: A mixed methods study of the views of affected families.

Autosomal recessive conditions are a significant health burden with few treatments. Population carrier screening has been suggested as a means to tackle them. Little is known about the views of affected families despite the potential for direct impacts on them. Data are presented on attitudes among families affected by Spinal Muscular Atrophy (SMA) toward two population screening programs, pre-conception, and prenatal. Data were gathered through qualitative interviews (n = 36) and a survey (n = 337). Eighty-two survey participants had SMA and 255 were family members. The majority were in favor of screening (75%). Reasons for supporting pre-conception screening support were a belief that it would reduce SMA-related terminations and raise awareness of SMA in the population. For prenatal screening, reasons for support included a belief in the importance of informed decision-making and the need to reduce suffering. Key reasons for non-support of pre-conception screening included concerns about carrier stigmatization and social engineering. For prenatal screening, concerns focused on the collateral loss of high quality of life lives affected by SMA. This study highlights that those affected by SMA are predominantly in favor of screening, although pre-conception screening is most favored. While family members and adults with SMA had largely consistent views, perceptions varied according to the severity (type) of SMA, with those affected by SMA type II the least likely to support screening. These findings suggest that screening for SMA is a complex issue for affected families, underscoring the need to consider and include their views when planning and implementing screening programs. © 2016 Wiley Periodicals, Inc.

Femtosecond Study of Dimolybdenum Paddlewheel Compounds with Amide/Thioamide Ligands: Symmetry, Electronic Structure, and Charge Distribution in the 1MLCT S1 State.

Four photophysically interesting dimolybdenum paddlewheel compounds are synthesized and characterized: I and II contain amide ligand (N,3-diphenyl-2-propynamide), and III and IV contain thioamide ligand (N,3-diphenyl-2-propynethioamide). I and III are trans-Mo2L2(O2C-TiPB)2-type compounds, and II and IV are Mo2L4-type compounds, where O2C-TiPB is 2,4,6-triisopropylbenzoate. I-IV display strong light absorption due to metal to ligand charge transfer (MLCT) transitions from molybdenum to the amide/thioamide ligands. Charge transfer dynamics in the MLCT excited states of I-IV have been examined using femtosecond transient absorption (fs-TA) spectroscopy and femtosecond time-resolved infrared (fs-TRIR) spectroscopy. The asymmetric amide/thioamide ligands show two forms of regioarrangements in the paddlewheel compounds. Analyses of the ν(C≡C) bands in the fs-TRIR spectra of I and II show similar electron density distribution over ligands in their 1MLCT S1 states where only two amide ligands are involved and the transferred electron is mainly localized on one of them. The fs-TRIR spectra of III and IV, however, show different charge distribution patterns where the transferred electron is fully delocalized over two thioamide ligands in III and partially delocalized in IV. Fast interligand electron transfer (ILET) was recognized as the explanation for the various charge distribution patterns, and ILET was shown to be influenced by both the ligands and the ligand arrangements.

Probing Interligand Electron Transfer in the 1MLCT S1 Excited State of trans-Mo2L2L'2 Compounds: A Comparative Study of Auxiliary Ligands and Solvents.

The interligand charge dynamics of the lowest singlet metal-to-ligand charge-transfer states (1MLCT S1 states) of a series of quadruply bonded trans-Mo2(NN)2(O2C-X)2 paddlewheel compounds are investigated, where NN is a π-accepting phenylpropiolamidinate ligand and O2C-X (X = Me, tBu, TiPB, or CF3) is an auxiliary carboxylate ligand. The compounds show strong light absorption in the visible region due to MLCT transitions from the Mo2 center to the NN ligands. The transferred electron density was followed by femtosecond time-resolved infrared (fs-TRIR) spectroscopy with vibrational reporters such as the ethynyl groups on the NN ligands. The observed fs-TRIR spectra show that these compounds have asymmetric 1MLCT S1 excited states where the transferred electron mainly resides on a single NN ligand. The presence of interligand electron transfer (ILET) is suggested to explain the shape of the ν(C≡C) bands and the influence of auxiliary ligands and solvents on the interligand electronic coupling. The ILET in the 1MLCT S1 state is shown to be sensitive to the functional groups on the auxiliary ligands while being less responsive to changes in solvents.

Synthesis, Structure, and Photophysical Properties of Mo2(NN)4 and Mo2(NN)2(T(i)PB)2, Where NN = N,N'-Diphenylphenylpropiolamidinate and T(i)PB = 2,4,6-Triisopropylbenzoate.

Two dimolybdenum compounds featuring amidinate ligands with a C≡C bond, Mo2(NN)4 (I), where NN = N,N'-diphenylphenylpropiolamidinate, and trans-Mo2(NN)2(T(i)PB)2 (II), where T(i)PB = 2,4,6-triisopropylbenzoate, have been prepared and structurally characterized by single-crystal X-ray crystallography. Together with Mo2(DAniF)4 (III), where DAniF = N,N'-bis(p-anisyl)formamidinate, all three compounds have been studied with steady-state UV-vis, IR, and time-resolved spectroscopy methods. I and II display intense metal to ligand charge transfer (MLCT). Singlet state (S1) lifetimes of I-III are determined to be 0.7, 19.1, and 2.0 ps, respectively. All three compounds have long-lived triplet state (T1) lifetimes around 100 µs. In femtosecond time-resolved infrared (fs-TRIR) experiments, one ν(C≡C) band is observed at the S1 state for I but two for II, which indicate different patterns of charge distribution. The electron would have to be localized on one NN ligand in I and partially delocalized over two NN ligands in II to account for the observations. The result is a standard showcase of excited-state mixed valence in coordination compounds.

Electronic and Spectroscopic Properties of Avobenzone Derivatives Attached to Mo2 Quadruple Bonds: Suppression of the Photochemical Enol-to-Keto Transformation.

From the reactions between Mo2(T(i)PB)4, where T(i)PB is 2,4,6-triisopropylbenzoate, and 2 equiv of the acids 4-formylbenzoic acid, HBzald; 4-(3-oxo-3-phenylpropanoyl)benzoic acid, HAvo; and 4-(2,2-difluoro-6-phenyl-2H-1λ(3),3,2λ(4)-dioxaborinin-4-yl)benzoic acid, HAvoBF2, the compounds Mo2(T(i)PB)2(Bzald)2, I; Mo2(T(i)PB)2(Avo)2, II; and Mo2(T(i)PB)2(AvoBF2)2, III, have been isolated. Compounds I and II are red, and compound III is blue. The new compounds have been characterized by (1)H NMR, MALDI-TOF MS, steady-state absorption and emission spectroscopies, and femtosecond and nanosecond time-resolved transient absorption and infrared spectroscopies. Electronic structure calculations employing density functional theory and time-dependent density functional theory have been carried out to aid in the interpretation of these data. These compounds have strong metal-to-ligand charge transfer, MLCT, and transitions in the visible region of their spectra, and these comprise the S1 states having lifetimes ∼5-15 ps. The triplet states are Mo2δδ* with lifetimes in the microseconds. The spectroscopic properties of I and II are similar, whereas the planarity of the ligand in III greatly lowers the energy of the MLCT and enhances the intensity of the time-resolved spectra. The Mo2 unit shifts the ground state equilibrium entirely to the enol form and quenches the degradation pathways of the avobenzone moiety.

Electronic and spectroscopic properties of avobenzone derivatives attached to ditungsten quadruple bonds.

From the reactions between W2(T(i)PB)4, where T(i)PB is 2,4,6-triisopropylbenzoate, and 2 equiv of acid, 4-formylbenzoic acid, HBzald, 4-(3-oxo-3-phenylpropanoyl)benzoic acid, HAvo, or 4-(2,2-difluoro-6-phenyl-2H-1λ(3),3,2λ(4)-dioxaborinin-4-yl)benzoic acid, HAvoBF2, three new compounds W2(T(i)PB)2(Bzald)2, I, W2(T(i)PB)2(Avo)2, II, and W2(T(i)PB)2(AvoBF2)2, III, have been prepared. As solid compounds I and II are blue while compound III is green. Characterization of these compounds has been carried out by means of (1)H NMR, MALDI-TOF MS, steady-state absorption and emission spectroscopies, and femtosecond and nanosecond transient absorption and time-resolved infrared spectroscopies. Compounds I and II have strong metal to ligand charge transfer, MLCT, transitions in the visible region of their spectra while compound III exhibits MLCT absorption in the near-infrared (λmax = 1017 nm). All three have S1 states that have corresponding lifetimes of ∼3-30 ps and are (1)MLCT in character. The triplet states are (3)MLCT with lifetimes in the range 3-10 ns. Density functional theory and time-dependent density functional theory were employed to perform electronic structure calculations in order to aid in the interpretation of these data. The spectroscopic properties of I and II are similar while the planarity of the ligand in III greatly lowers the energy of the MLCT state. The W2 unit enables direct observation of intersystem crossing from the (1)MLCT state to (3)MLCT state via the use of ultrafast spectroscopy.

Mo2 paddlewheel complexes functionalized with a single MLCT, S1 infrared-active carboxylate reporter ligand: preparation and studies of ground and photoexcited states.

From the reactions between Mo2(DAniF)3pivalate (DAniF = N,N'-di(p-anisyl)formamidinate) and the carboxylic acids LH, the title compounds Mo2(DAniF)3L have been prepared and characterized: compounds I (L = O2CC≡CPh), II (L = O2CC4H2SC≡CH), and III (L = O2CC6H4-p-CN). The new compounds have been characterized in their ground states by spectroscopy ((1)H NMR, ultraviolet-visible absorption, near-infrared absorption, and steady state emission), cyclic voltammetry, and density functional theory calculations. The compounds show strong metal Mo2 to ligand L δ-π* transitions in their visible spectra. The nature of the S1 (1)MLCT and T1 states has been probed by time-resolved (femtosecond and nanosecond) transient absorption and infrared spectroscopy. The observed shifts of the C≡C and C≡N vibrational modes are found to be consistent with the negative charge being localized on the single L in the S1 states, while the T1 states are (3)Mo2 δδ*. The present results are compared to earlier studies of the photoexcited states of trans-Mo2(2,4,6-triisopropylbenzoate)2L2 compounds that have been assigned as either localized or delocalized.

SMN and the Gemin proteins form sub-complexes that localise to both stationary and dynamic neurite granules.

Childhood spinal muscular atrophy (SMA) is caused by a reduction in survival motor neuron (SMN) protein. SMN is expressed in every cell type, but it is predominantly the lower motor neurones of the spinal cord that degenerate in SMA. SMN has been linked to the axonal transport of beta-actin mRNA, a breakdown in which could trigger disease onset. It is known that SMN is present in transport ribonucleoproteins (RNPs) granules that also contain Gemin2 and Gemin3. To further characterise these granules we have performed live cell imaging of GFP-tagged SMN, GFP-Gemin2, GFP-Gemin3, GFP-Gemin6 and GFP-Gemin7. In all, we have made two important observations: (1) SMN granules appear metamorphic; and (2) the SMN-Gemin complex(es) appears to localise to two distinct subsets of bodies in neurites; stationary bodies and smaller dynamic bodies. This study provides an insight into the neuronal function of the SMN complex.

Analysis of SMN-neurite granules: Core Cajal body components are absent from SMN-cytoplasmic complexes.

Childhood spinal muscular atrophy (SMA) is caused by a reduction in survival motor neuron (SMN) protein. SMN is a ubiquitously expressed house keeping protein that is involved in RNA production and processing. However, although SMN is expressed in every cell type, only the lower motor neurons of the spinal cord are degraded in SMA. It remains unclear why this is the case. Recently, SMN has been linked to the axonal transport of beta-actin mRNA from the cell body down to the growth cones. beta-Actin is transported actively in neurite granules (NGs). However, it remains unclear which known SMN-binding partners are present in these SMN-NGs. To address this we have analysed SMN-NGs in a human neuronal cell line, SH-SY5Y, using antibodies against the majority of reported SMN-binding partners, including: Gemin2, Gemin3, Gemin4, Gemin5, Gemin6, Gemin7, Sm core proteins, fibrillarin, EWS, PFNII, Unrip and ZPR1. The obtained results highlight the metamorphic nature of the SMN complex, suggesting that not all the "core" SMN-binding proteins are transported in SMN-NGs.

Joining the dots: production, processing and targeting of U snRNP to nuclear bodies.

The spliceosome is the RNP complex than catalyses the removal of introns from the Uridine-rich small nuclear ribonucleoproteins (U snRNPs) that make up the main components of this complex. The production of these RNPs is an intricate process, involving several key stages. These include: 1) the transcription of the U snRNAs; 2) their nuclear export; 3) the cytoplasmic assembly of the U snRNP; 4) their nuclear import; 5) their processing within Cajal bodies and the nucleolus; and 6) their storage in interchromatin granule clusters (IGCs). This review focuses on each of these stages, discussing the key complexes involved as well as the trafficking and targeting mechanisms involved.

Identification of a tripartite import signal in the Ewing Sarcoma protein (EWS).

The Ewing Sarcoma (EWS) protein is a ubiquitously expressed RNA processing factor that localises predominantly to the nucleus. However, the mechanism through which EWS enters the nucleus remains unclear, with differing reports identifying three separate import signals within the EWS protein. Here we have utilized a panel of truncated EWS proteins to clarify the reported nuclear localisation signals. We describe three C-terminal domains that are important for efficient EWS nuclear localization: (1) the third RGG-motif; (2) the last 10 amino acids (known as the PY-import motif); and (3) the zinc-finger motif. Although these three domains are involved in nuclear import, they are not independently capable of driving the efficient import of a GFP-moiety. However, collectively they form a complex tripartite signal that efficiently drives GFP-import into the nucleus. This study helps clarify the EWS import signal, and the identification of the involvement of both the RGG- and zinc-finger motifs has wide reaching implications.

Preventing lives affected by hemophilia: A mixed methods study of the views of adults with hemophilia and their families toward genetic screening.

BACKGROUND: Genomic sequencing technologies have made the possibility of population screening for whole panels of genetic disorders more feasible than ever before. As one of the most common single gene disorders affecting the UK population, hemophilia is an attractive candidate to include on such screening panels. However, very little is known about views toward genetic screening amongst people with hemophilia or their family members, despite the potential for a wide range of impacts on them. METHODS: Twenty-two in-depth qualitative interviews were undertaken to explore the views of adults with hemophilia and their family members, recruited through the Haemophilia Society UK. These interviews were used to develop a survey, the Haemophilia Screening Survey (UK), which was distributed in paper and online format through the support group, receiving 327 returns between January and June 2018. RESULTS: Fifty-seven per cent of the sample supported preconception carrier screening of the population for hemophilia, and 59% supported prenatal carrier screening. Key reasons for support included a desire to reduce pregnancy terminations and increase awareness of hemophilia. Despite support for screening however, 90% of the sample disagreed with pregnancy terminations for hemophilia. CONCLUSIONS: Families and adults living with hemophilia are more supportive of screening for information and preparation purposes than to prevent boys with hemophilia from being born. A distinction was made between preventing the disease and preventing the lives of people with it, with support shown for the use of screening to achieve the former, but not at the expense of the latter.

Identification and characterisation of a nuclear localisation signal in the SMN associated protein, Gemin4.

Gemin4 is a ubiquitously expressed multifunctional protein that is involved in U snRNP assembly, apoptosis, nuclear/cytoplasmic transportation, transcription, and RNAi pathways. Gemin4 is one of the core components of the Gemin-complex, which also contains survival motor neuron (SMN), the seven Gemin proteins (Gemin2-8), and Unrip. Mutations in the SMN1 gene cause the autosomal recessive disorder spinal muscular atrophy (SMA). Although the functions assigned to Gemin4 predominantly occur in the nucleus, the mechanisms that mediate the nuclear import of Gemin4 remain unclear. Here, using a novel panel of Gemin4 constructs we identify a canonical nuclear import sequence (NLS) in the N-terminus of Gemin4. The Gemin4 NLS is necessary and independently sufficient to mediate nuclear import of Gemin4. This is the first functional NLS identified within the SMN-Gemin complex.