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Jones et Al. are investigating the effect of 3 compounds on SMN2 expression, a gene involved in Spinal Muscular Atrophy.  From the Fig. 1, graphs, we can observe that increasing concentration of 3 compounds, SMN-C1, SMN-C2, and SMN-C3, leads to higher FL SMN2 mRNA transcript level and a lower Δ7  SMN2 mRNA transcript level expression.  Such statement relates back to the banding patterns seen above, showing similar FL and Δ7 expression at a low SMN-C1 concentration (3nM) but a much higher FL expression and lower/absent Δ7 expression at 1 μm (highest concentration). This suggests inadequate RNA splicing decreases in a dose-dependent manner. Interestingly, in figure 2, we can observe 6 genes are respectively up and down regulated after SMNC-3 treatment. While SM2 expression fold change value increases, the latter increase is not significant. We may hence argue that SMNC-3 affects only the way SM2 is spliced but not significantly the amount of gene expression itself. These 12 genes could as well cause the change we observe in splicing tendencies in Fig.1. According to Figure 3a, we can see that plasma and brain SNM-C3 concentration increase about 2.5/3 hours (brain SNM-C3 being higher), following by a decreasing trend from this point on, showing that the drug can access the brain. Hence, SNM-C3 has an impact on SMN protein expression, as further outlined by Figure 3b, in which the percentage change from baseline is increased in the event of day 1 and day 10 injections and decreases abruptly back to normal after 168 hours. This aspect emphasizes on the unstable nature of the SMN protein. These overall observations provide a good glimpse of how SMN-C1, C2, and C3 can constitute a therapeutic target, namely by lowering alternative splicing and promoting SMN expression (here, by SMN-C3), but brings about a few risks: the 6 respective up and down regulated gene could code for specific proteins promoting oncogenic processes. Secondly, continuous SMN-C3 treatment could potentially heighten SMN doses at dangerously high levels, observation which, combined with the fact that the mouse model system is much different than the human’s, constitute crucial issues.

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The first experiment done was treating cells from SMA patients with three different compounds; SNM-C1, SNM-C2, SNM-C3. The levels of mRNA transcript of the spliced SMN2 (disease-causing mRNA transcript) and levels of the normal SMN2 transcript were measured using RT-PCR. The graphs presented showed that all of the compounds tested, can decrease the transcription of the disease-causing alternative spiced version of the SNM2 mRNA transcript (Delta7 SNM2).

Then, Jones et al. looked more closely and the regulation of gene expression when SMA patients are treated with SMA-C3. By comparing to a control of SMA patients treated with DMSO, the treatment with SMA-C3 does not have a significant on most genes (within the fibroblasts) transcription levels. However, 7 genes were upregulated and 7 were downregulated with SMA-C3 treatment. Looking specifically at the SMA2 gene, it’s expression was not affected by SNM-C3, suggesting that the normal expression of the gene is not affected, but comparing it to the previous figure, SMN-C3 only changes the way that SMN2 is spliced and not overall the gene expression.

A mouse was used as an in vivo model to give Jones et al. an idea of specifically SNM-C3's effect on a larger scale, compared to just looking at fibroblasts. The treatment goes to the mice brain in the first place, as the plasma concentration of SNM-C3 in the brain rises only a few hours after the dose. This is beneficial as the drug can signal to the rest of the body to treat the disease.

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Hence, Jones et al. applied three different compounds to both fibroblasts derived from SMA patients and a mouse model, and then derived 3 figures from the measurements they took. The first figure shows that, in term of SMN2 DNA sequence in the fibroblasts, the full length increased and the alternatively form missing exon 7 decrease as the concentrations of all three compounds increased. From this change, the SMN-C1, C2 and C3 all shows a inhibition effect on RNA splicing. From the second figure, nearly equal number of genes were upregulated and downregulated by SMN-C3 treatment, and SMN2 transcription is unaffected, suggest that the genes that have altered expression may regulate the SMN2 splicing. The last figure demonstrate that, the concentration of SMN-C3 in both plasma and brain increased steadily within the first three hours after it was given, and that in brain is more significant that in plasma. Furthermore, the SMN protein level increased remarkedly, and peak around 250% within 24 hours after the SMN-C3 treatment. However, in term of SMN protein, though the increase for brain is faster and higher, the decline is quicker as well. Overall, the SMN-C3 function nicely during the experiment, but some other important factors still wait to be tested, for example, the other organs besides brain have not be tested yet; Also, it is still based on a mouse model, the larger human size and different RNA splicing mechanism are not took into account either.

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Hence, Jones et al. applied three different compounds to both fibroblasts derived from SMA patients and a mouse model, and then derived 3 figures from the measurements they took. The first figure shows that, in term of SMN2 DNA sequence in the fibroblasts, the full length increased and the alternatively form missing exon 7 decrease as the concentrations of all three compounds increased. From this change, the SMN-C1, C2 and C3 all shows a inhibition effect on RNA splicing. From the second figure, nearly equal number of genes were upregulated and downregulated by SMN-C3 treatment, and SMN2 transcription is unaffected, suggest that the genes that have altered expression may regulate the SMN2 splicing. The last figure demonstrate that, the concentration of SMN-C3 in both plasma and brain increased steadily within the first three hours after it was given, and that in brain is more significant that in plasma. Furthermore, the SMN protein level increased remarkedly, and peak around 250% within 24 hours after the SMN-C3 treatment. However, in term of SMN protein, though the increase for brain is faster and higher, the decline is quicker as well. Overall, the SMN-C3 function nicely during the experiment, but some other important factors still wait to be tested, for example, the other organs besides brain have not be tested yet; Also, it is still based on a mouse model, the larger human size and different RNA splicing mechanism are not took into account either.

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Effect of SMN-C1~3 on the level of functional SMN

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Effect of SMN-C1~3 on the level of functional SMN

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From this experiment, molecules SMN-C1,C2,C3 all showed an elevating effect in the level of functional SMN2. This effect was not achieved through the elevation of expression of SMN2 gene as there is no significant change in the total mRNA abundance of SMN2 gene. Instead, SMN-c1~3b exhibited an ability in shifting the preference of splicing from the form lacking exon 7 to the functional form. As the concentration of SMN-C1~3 applied to fibroblasts increases, the level of SMN2 lacking exon decreases, which is the reverse of functional SMN2.

Jones et al. then used an mouse model to examine the impact of SMN-C1~3 in vivo. Once daily oral doses of SMN-C3 (10 mg/kg) was given to mice and the concentration of SMN-C3 and functional SMN was measured on day 1 and 10. The level of SMN quickly increased in 12 hours then wear off, which matches the trend of concentration of SMN-C3 in mouse. However, SMN-C3 did exhibit an accumulating influence on the level of SMN. The peak concentration of SMN after intake of SMN-C3 on day 10 was two times higher than the peak concentration on day 1.

Overall, SMN-C1~3 seems like a potential treatment for SMA, but treatment must be long-term to maintain sufficient level of SMN. However, SMN-C1~3 did impacted the expression level of around 15 other genes in fibroblasts which must be looking into for following consequences. And the effect of SMN-C1~3 on splicing mechanisms of other genes was not examined in the experiment.

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After applying three compounds, SMN-C1, SMN-C2 and SMN-C3, to fibroblasts derived from SMA patients and quantifying the full length (FL) SMN2 and alternatively spliced form missing exon 7 (∆7) using RT-PCR (see figure 1), the data collected showed very interesting results. As the compound concentration increased, relative transcript level of FL mRNA increased exponentially, while the relative transcript level of ∆7 level decreased exponentially. This data demonstrated that those compounds can modify the mRNA precursor to the SMN2 gene, as alternative splicing is decreased when in presence of the compounds.

In vivo models were then investigated, where SMA mice were given oral doses of the SMN-C3 (see figure 3). Plasma and brain tissues were collected and the compound concentrations were quantified over a 24 hour period. The results showed a spike in concentration in the first 6 hours, followed by a decrease. When quantifying SMN protein in quadriceps and brain after once daily dose on days 1 and 10, it was clear that the level of SMN protein was higher in day 10 than day 1.

The data discussed above shows a great potential for those compounds to offer a new potential treatment for spinal muscular atrophy (SMA).

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Three compunds, SMN-C1, SMNC2 and SMN-C3 were selected and their effects were tested on fibroblasts derived from SMA patients. Jones et al. quantified the transcription level of full length (FL) SMN2 and the alternatively spliced form missing exon 7 (Δ7) mRNA, and found that with increasing concentration (0.01μM, 0.1μM, and 1μM) of SMN-C1, SMN-C2 and SMN-C3, the transcription level of FL mRNA increased and that of Δ7 mRNA decreased. This suggests that all 3 molecules are able to pharmacologically reduce alternatively splicing of SMN2 gene transcripts.
In the patient fibroblasts used, 11,714 genes are found to be expressed. In a second experiment, upon treatment of SMN-C3, about 6 genes were upregulated and 6 were downregulated, but the transcription level of SMN2 was unaffected. This suggests that one of those genes whose expression level was changed probably had an effect on the splicing process of SMN2 gene.
They then looked at a mouse model for SMA that was given daily oral doses (10 mh/kg) of SMN-C3 for 10 days. They found that there was an initial rise in SMN-C3 concentration in plasma and brain right after the dose was given and then decrease over time, with much higher concentration in the plasma than the brain. It is key that this confirms SMN-C3 can penetrate into the brain. Jones et al. then measured the level of SMN proteins in the quadriceps and brain in vivo. The data is clear that after 10 days, the amount of SMN protein was much higher in the mouse compared to that in day 1, so suggesting that with the accumulation of SMN-C3 in the body, a higher SMN protein level can be achieved.

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In the article, SMN-C3 is presented as a possible novel drug to treat SMA. SMN-C3 showed to increase in the amount of full-length SMN2 transcript (the healthy version). Consequently, the alternatively spliced form missing exon 7 was reduced.

The mechanism by which SMN-C3 is increased is not entirely known, but there was upregulation and downregulation of certain genes. It could be possible that those genes could affect the splicing mechanism. Finally, it was shown that at higher concentrations of SMN-C3 there were higher percentages of healthy SMN protein.

However, the results should be taken with a pinch of salt. Firstly because the experiments were done in mice models. Therefore, it is not entirely possible to predict the consequences of SMN-C3 on humans. Secondly, SMN-C3 intake changed the expression of several genes. Although they could be involved in the splicing mechanism, they could also produce unwanted side effects.

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Jones et al. investigates three compounds SMN-C1, SMN-C2 and SMN-C3 on treating SMA. WhenSMN-C1, SMN-C2 and SMN-C3 were applied to fibroblasts derived from SMA patients. The experimental data demonstrates when the concentration of these three compounds increases (from 0.01uM to 1uM), the full length mRNA increased and the level of alternative RNA spliced, then the exon7 is lowered.

SMN-C3 has no significant change in total mRNA abundance. This suggests one of the gene did have altered expression could be mediating the change in SMN2 splicing detected.

A mouse model with SMA was also investigated by given oral doses of SMN-C1, SMN-C2 and SMN-C3 (10mg/kg) for 10 days. Within a few hours after the dose, the SMN protein level at both quadriceps and brain increases significantly. The level of SMN protein was then degrades gradually. The difference between day 1 and day 10 is that the overall level of SMN protein increases and the speed of SMN degradation slowed down. We can conclude that the three SMN compounds can affect the alternative splicing and could be a possible treatment for the SMA. However, side effects may occur as the SMN-C3 alters the splicing of 12 different genes shown in the second figure.

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Three compounds (SMN-C1, SMN-C2 and SMN-C3) were treated with the fibroblasts derived from the SMA patient. All 3 compounds have shown to be altering the transcription level of both full length SMN2 and its alternatively spliced form with missing exon 7, as suggested from the RT-PCR analysis. The 3 compounds helped increasing the transcription of the full length mRNA, and repressed the transcription of the alternatively spliced form, as their concentrations increased; they could fully repress the transcription of the alternatively spliced form at a very high concentration. SMN-C3 in particular, did not have a significant effect on the splicing in fibroblasts, as within the fibroblasts only 14 out of the 11,714 genes were up-regulated or down-regulated by SMN-C3. One of these altered genes might have side-effect that mediate the change in SMN2 splicing instead of SMN-C3, as SMN2 did not have a significant change in mRNA abundance when treated with SMN-C3. From an in vivo model, SMN-C3 concentration decreased as time passed after the oral dose, showing SMN protein was an unstable protein. More specifically, the brain and quadricep absorbed more SMN-C3 than the plasma, and there was an accumulative effect of SMN protein when doses were administered more regularly.

SMN compounds have shown to upregulate the full length gene and downregulate the alternatively form, so they could be a potential therapeutic target. However, SMN-C3 showed to have significant effect on 14 off-target genes imposing a high risk as the brain has a high absorption. Further studies needed to investigate the associated risk by the off-targeting effect on genes.

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Jones et al. investigate the effects of three compounds SMN-C1, SMN-C2, SMN-C3 on the SMN2 gene. Figure 1 shows that after treating with 1µM of all three compounds, the relative transcription level of full length SMN2 increase while the transcription level of alternatively spliced form missing exon 7 (∆7) decreases to almost 0. This indicates that all three molecules reduce RNA splicing.

Figure 2 shows that 7 genes are upregulated, and 7 genes are downregulated by SMN-C3, while SMN2 is not significantly affected. Those genes that have an altered expression level could potentially affect the splicing of SMN2 giving the result of Figure 1.

From in vivo model, it is suggested that SMN-C3 could actually reach the brain and have an effect over SMN protein level in living organisms. Also, the fact that SMN protein level gradually decreases after the day 10 dose indicates that the protein is not very stable and decomposes overtime. This study suggests potential treatment for SMA. However, the fact that 7 genes were upregulated and 7 genes were downregulated posts a potential threat to the body.

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The study by Jones team al revealed that injected compounds SMN-C1, SMN-C2 and SMN-C3 tend to affect the proportions of mRNA transcript existed in the fibroblasts from SMA patients. PCR is used to demonstrate how different types of SMN-C molecules will affect lengths of mRNA and this shows the alternative splicing is decreased by adding SMN molecules.Figure 2 illustrates there is no clear up or downstream regulatory patterns following SMN-C3 treatment as the lengths of transcripts did not exceed the two lines of fold changes too frequently. It could be one gene is changed and cause those few genes up and downstream regulated. Figure 3 shows that in the brain the SMN levels increase dramatically at first but also fasten at a higher rate than in the plasma. It proves that SMN protein can get into brain and compared the one in day10, the protein level in day1 degrades fat a faster rate. In their essay, we analyse that SMNcan affect the level of alternative splicing

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In the study, all of SMN-C1,C2, and C3 are making longer full length mRNA for SMN2 gene. They cause the change in exon 7 to decrease compared to the mutated one, which making the mRNA level closer to the normal standard. As a result, the RNA splicing will become normal and stop splicing the exon 7 out. For the transcriptional changes by 3 compounds, its showing that SMN2 is not significantly changed, while there are several other genes fluctuating above doubling line or below half line. The resulting changes balanced out the splicing of exon 7. However, it is not caused by SMN2 but by other genes that do change. Next, the mouse model shows that SMN compounds do enter the plasma, brain, and quadriceps, especially high in brain at the first 24 hours and it takes much longer time for mouse to metabolize SMN proteins after taking 10 days’ oral doses than only 1 day’s oral dose. 

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In the study, all of SMN-C1,C2, and C3 are making longer full length mRNA for SMN2 gene. They cause the change in exon 7 to decrease compared to the mutated one, which making the mRNA level closer to the normal standard. As a result, the RNA splicing will become normal and stop splicing the exon 7 out. For the transcriptional changes by 3 compounds, its showing that SMN2 is not significantly changed, while there are several other genes fluctuating above doubling line or below half line. The resulting changes balanced out the splicing of exon 7. However, it is not caused by SMN2 but by other genes that do change. Next, the mouse model shows that SMN compounds do enter the plasma, brain, and quadriceps, especially high in brain at the first 24 hours and it takes much longer time for mouse to metabolize SMN proteins after taking 10 days’ oral doses than only 1 day’s oral dose. 

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In the Jones et al study it indicates that the compounds SMN-C1, SMN-C2, and SMN-C3 can all increase the levels of SMN protein in healthy non mutant mRNA (FL mRNA). The three drugs were applied to the fibroblasts that were derived the cells of SMA patients to see if they could modify the mRNA precursor to the gene SMN2 (survival of motor neuron 2). These compounds could therefore possibly increase the level of functional SMN2 and therefore begin to alleviate or stop the symptoms of SMN2. Through the paper the compounds were investigated. When exon 7 was removed, as shown in figure, from the mRNA the effects of the three compounds, that were injected in a dose dependant manner, decreased causing the relative transcript level to increase and therefore causing alternative splicing to decrease. The analysis of SMN-C3 in figure 2, revealed that there was no significant change in the abundance of mRNA, but 6 genes were up and down regulated therefore suggesting that one of these genes may have affected the splicing of SMN2. Treatment with a living mouse in figure 3 of C3 led to an increase in brain and quadricep SMN2 levels for around 40 hours before the treatment efficacy decreased as levels began to return to normal. The impact of this paper, shows that C3 may have the potential to be a target for therapeutical medicines for SMA. However SMN-C3, also showed some other unwanted effects that include a build-up of SMN in the body (figure 3 – day 10) which took longer for SMN to be removed from the body of the mouse in comparison to day 1. Also the treatment caused an affect in the brain which is not expected.

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An initial experiment was conducted in which cells containing one of the two forms of the SMN2 gene  were treated for 24 hours, with three different compounds; SMN-C1, SMN-C2 and SMN-C3, followed by RT-PCR analysis of full length (FL) SMN2 and the alternatively spliced form missing exon 7 (△7). This allowed for the comparison of the relative transcription levels of both forms after treatment with the different compounds at different concentrations between 0uM and 1uM. SMN-C1 showed the greatest increase in relative transcription level of the FL SMN2 gene and was also shown to decrease relative transcription level of the △7 SMN2 gene. All three compounds showed the same trend. This trend was further supported by a gel that was conducted on cells treated with SMN-C1.

A measure of the difference in total transcription expression of 11,714 genes was then conducted to identify if the expression of any genes was affected following SMN-C3 treatment. SMN-C3 was shown to have no significant effect on the expression of the SMN2 gene, however it was shown to have a significant effect on the expression of 12 (0.001%) genes, with half of these being significantly upregulated and the other half being significantly downregulated.

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In her study, Jones. tested the effect of three different compounds: SMN-C1, SMN-C2 and SMN-C3 on SMN splicing in vitro and then in various tissues using a mouse model. What she found was that when introduced outside of an organism all three compounds show potential to increase the levels of healthy SMN protein and reduce the translation of the faulty delta7 variant. (fig.1). SMN-C2 seems the most reliable candidate as it does not lower the overall translation of SMN the way SMN-C3 does and it effects are observable on a shorter timescale than SMN-C1. When replicated in vivo, treating mice with a dose of the potential drug indeed increased plasma levels of the healthy NMS molecules and so it did in brain and muscle tissue showing that the drug is capable of targeting neurons. As demonstrated by fig.2 this change in the absolute levels of SMN protein and not due to an overall change in the amount of gene transcription going on but a correction to the structure of SMN. Receiving SMN-C2 daily induced an ability to retain altered levels of SMN proteins for up to 168 after 10 days of daily treatment.

If SMN-C2 can demonstrate clinically significant improvement of motor neurons' function it could become a potential treatment for Spinal Muscular Atrophy. However, the study demonstrated that SMN-C2 dysregulates the expression of about 12 genes other than SMN so it is difficult just yet to determine the net effect this would have in patients and further investigations are needed to establish the exact relationship between SMN-C2 administration and motor neurons function as well as address any safety concerns such as off-target effects the treatment might cause in other genes and aspects of physiology.

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Based on the results, the 3 SMN molecules have similar enhancing effects for transcription of FL and similar inhibitory effects for transcription of delta-7, with SMN-C1 being the most enhancing and the least inhibitory while SMN-C3 being the least enhancing and the most inhibitory. These effects are more amplified at higher concentrations. This shows that SMN molecules are effective to lower the transcription of delta-7 SMN (forming the mutant protein) and increase transcription of the normal SMN gene to form the normal protein, with SMN-C3 being the most effective.

However, with SMN-C3, the amount of SMN2 gene is not significantly affected compared with 11713 other genes. However, with repeated administration of SMN-C3, SMN protein is increasingly expressed due to increasing start concentrations of SMN proteins. In day 10 of administration, SMN protein levels are still 25% higher than the baseline after 168 hours, which shows that SMN-C3 helps the production of normal SMN protein (which is not rapidly degraded). These data collectively show the positive correlation of SMN-C3 dosage and normal SMN protein levels, which may suggest that SMN-C3 is a feasible drug for the disease.

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Based on the results, the 3 SMN molecules have similar enhancing effects for transcription of FL and similar inhibitory effects for transcription of delta-7, with SMN-C1 being the most enhancing and the least inhibitory while SMN-C3 being the least enhancing and the most inhibitory. These effects are more amplified at higher concentrations. This shows that SMN molecules are effective to lower the transcription of delta-7 SMN (forming the mutant protein) and increase transcription of the normal SMN gene to form the normal protein, with SMN-C3 being the most effective.

However, with SMN-C3, the amount of SMN2 gene is not significantly affected compared with 11713 other genes. However, with repeated administration of SMN-C3, SMN protein is increasingly expressed due to increasing start concentrations of SMN proteins. In day 10 of administration, SMN protein levels are still 25% higher than the baseline after 168 hours, which shows that SMN-C3 helps the production of normal SMN protein (which is not rapidly degraded). These data collectively show the positive correlation of SMN-C3 dosage and normal SMN protein levels, which may suggest that SMN-C3 is a feasible drug for the disease.

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SMN-C1, SMN-C2 and SMN-C3 were used in the research. In the in vitro experiments it was shown that all of them played a role in increasing the relative transcript level in full length mRNA and reducing the relative transcript level in exon 7 missing mRNA after 24 hours of cell treatment. The effectiveness of all three compounds were similar which rose as compound concentration was increased. Jones et al. also examined the impacts of SMN-C3 on the level of gene expression in patient fibroblasts. It was indicated that 7 genes were up-regulated and 7 genes were down-regulated. However, there was no significant change in the transcription of SMN2 which means SMN-C3 reduced the percentage of mutated alternative splicing without changing the amount of SMN2 gene transcribed. Furthermore, in vivo experiments on mice proved that SMN-C3 mainly travelled to the brain after oral ingestion instead of plasma and the concentration of SMN-C3 dropped over 24-hour period after the intake. It also boosted the amount of SMN proteins and this effect accumulated over a 10 day period.

In conclusion, this research gives us some insights into the possibility of using SMN-C1, SMN-C2 and SMN-C3 to treat SMA patients in human. More researches might be done to investigate other compounds that can be effective on treating other diseases caused splicing errors. However, this treatments can be risky as experiments haven't carried out in human where the genome is different from the genome of mice and SMN-C1, SMN-C2 and SMN-C3 may interact with human genome in an unknown but possibly harmful way.

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Three different compounds were used in this investigation: SMN-C1, SMN-C2, and SMN-C3. Samples of fibroblast were first derived from SMA patients to explore whether different SMN2 treatments affected RNA splicing. The method RT-PCR is used to quantify relative levels of SMN2 RNA and exon 7 expression (Δ7), which are compared against the internal control GAPDH. Jones et al. showed that an increase in the concentration of all three compounds increased the concentration of full-length SMN2 mRNA and decreased the concentration of Δ7. The researchers have further investigated treatment changes involving compound SMN-C3. They have shown that a few genes have been upregulated and downregulated after the treatment, but the figure they have provided us has shown an insignificant change in SMN2 gene expression. The change in the other genes may have had an effect on the splicing mechanism of SMN2 mRNA. Moreover, they have given oral doses of SMN-C3 to mice and have collected brains and blood tissues from the mice to analyse the treatment effect. The data has shown that it takes about 5 hours for the treatment to take effect as the maximum concentration of SMN-C3 is reached in both brain and blood tissue. The treatment must also be taken over a couple of days to significantly increase the change in SMN protein percentage in the brain or quadriceps. Finally, the treatments, especially SMN-C3, has shown potential in treating SMA patients, but it must be noted that though the SMN2 gene has not shown a significant change in gene expression as previously suggested, it did cause a significant change in 12 of the other genes, which shows that there are risks involved with using the SMN-C3 treatment. Furthermore, the experiments were either performed in vitro or in mice (in vivo), not in humans, so the potential risks the treatment have on humans is inconclusive.

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Their research found that with an increase of SMN- C1, C2 and C3, transcription of exon 7 decreased, with higher concentrations resulting in complete lack of transcription. This suggests that all three forms of SMN affect alternative splicing and could lead to the absence of exon 7, therefore making them all potential contributors to the development of SMA. Despite its clear effect on splicing in relation to exon 7, SMN-C3 was not actually found to have a major overall effect on splicing in fibroblasts; of the 11,714 genes tested by Jones et al., only 14 showed a significant change in transcription (with 7 being upregulated and downregulated, respectively). These 14 genes could however, potentially be of interest in terms of SMA.

When Jones et al. administered SMN-C3 in vivo to a mouse model, it was seen that the C3 was absorbed in much higher concentrations in the brain compared to the plasma. This could be a concern, as this increases the likelihood of the motor neurons within the brain being affected and of an affected individual developing SMA. A higher concentration of SMN-C3 was seen in the quadriceps as compared to the brain, and this could again imply that the motor neurons within these muscles could be significantly affected in the disease.

These findings highlight that all three forms of SMN can be implicated within SMA and could be considered potential therapeutical targets in treatment of SMA. It would also be valuable to further research the 14 genes affected by SMN-3, to evaluate any other potential risks.

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Jones et al. applied three of the potential ‘magic compounds’ (SMN-C1, C2, C3) to SMA patient fibroblasts. Figure 1 shows the total level of expression for full length (FL) SMA and alternatively spliced SMA did not change due to an increase in SMN-C1, however, the expression of FL SMA increases while the alternatively spliced version decreases. Furthermore, as the amount of each compound increases, the relative transcript level of FL SMA increases. This could suggest the drugs prevent alternative splicing of the SMA gene. Figure 2 shows that there wasn’t a significant increase in gene expression of SMN2 because majority of the genes fall within the +1 and –1 bands.

From figure 3, we can see as time post-dose increases, less FL SMN2 is made and more alternative spliced SMN2 is made. Since the alternative spliced SMN2 is missing exon 7, it eventually becomes degraded which is why the level of SMN protein decreases. However, after 10 days of treatment the concentration of full length SMA protein increased significantly from baseline, and it took 168 hours for full length protein levels to decrease to normal. This implies that continuous use of the drug could lead to an increase in SMN protein production.

This research could prove to be useful in developing treatment for patients with SMA.

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Spinal muscular atrophy (SMA) is a genetic disease that mainly affects the central nervous system and leads to eventual degradation of the motor neurons found in the spinal cord. This is due to mutations in the survival motor neuron (SMN2), which codes for the SMN protein responsible for motor neuronal maintenance. The drugs, SMN-C1, C2 and C3, that are thought to artificially increase the functionality of the SMN2 gene.

Fibroblasts taken from SMA patients were treated with the aforementioned drugs and a PCR was carried out to see the levels of full length (FL) mRNA and mutated spliced SMN2 genes (∆7), found in the cells. It was found that increased SMN-C1 dosage contributed to a lower expression of mutated SMN2 proteins. Similarly, SMN-C2 and C3 showed a similar trend in the decrease of ∆7 transcription and an increase in FL transcription.

When tested in vivo, SMN C-3 was shown to increase the SMN protein expression in both the peripheral and central nervous system over a period of 10 days from the initial consumption. This could have adverse effects on the peripheral nervous system which was not the target region. The drug is therefore not a specific and targeted treatment which could cause neuronal damage in other unaffected areas of the body.

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With the fibroblasts derived from SMA patients, Jones et al. exanimated those tissue with three different compounds, SMN-C1, SMN-C2, SMN-C3. RT-PCR was applied to quantify the level of transcription with full-length SMN2 and the mutated SMN2 protein with no exon 7. The result had shown that all three compounds have the ability to promote the expression of normal SMN2 protein while suppressing the mutated SMN2 transcript. And with the investigation of SMN-C3 impacted gene transcription, results revealed that mRNA abundance remain roughly unchanged with some fluctuation, this shown that the downregulation of mutated SMN2 was compensated by the normal SMN2. After testing the drug in vitro, Jones et al. decided to perform an in vivo experiment by giving oral doses of SMN-C3 to mice daily and recording the level of the drug and SMN protein in the tissue. The results shown that there are more SMN-C3 detected in the brain than plasma, but the SMN protein is present more in quadriceps than in the brain. The SMN protein will accumulate since the prescription in day 10 causes a larger increase than on day 1 indicating that SMN protein will stay at a high level until the second dose. This could be a hazard since we don’t know whether a high level of SMN protein will harm the mouse.

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With the fibroblasts derived from SMA patients, Jones et al. exanimated those tissue with three different compounds, SMN-C1, SMN-C2, SMN-C3. RT-PCR was applied to quantify the level of transcription with full-length SMN2 and the mutated SMN2 protein with no exon 7. The result had shown that all three compounds have the ability to promote the expression of normal SMN2 protein while suppressing the mutated SMN2 transcript. And with the investigation of SMN-C3 impacted gene transcription, results revealed that mRNA abundance remain roughly unchanged with some fluctuation, this shown that the downregulation of mutated SMN2 was compensated by the normal SMN2. After testing the drug in vitro, Jones et al. decided to perform an in vivo experiment by giving oral doses of SMN-C3 to mice daily and recording the level of the drug and SMN protein in the tissue. The results shown that there are more SMN-C3 detected in the brain than plasma, but the SMN protein is present more in quadriceps than in the brain. The SMN protein will accumulate since the prescription in day 10 causes a larger increase than on day 1 indicating that SMN protein will stay at a high level until the second dose. This could be a hazard since we don’t know whether a high level of SMN protein will harm the mouse.

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Jones et al. administered 3 molecules, SMN-C1, C2 and C3, to fibroblasts derived from SMA patients, and observed the effects on RNA transcript and SMN protein level. It was observed that cells administered with 1um of any molecule showed an increase in transcript level for full length (FL) mRNA, while in ∆7 mRNA there was no relative transcript level recorded via RT-PCR. This suggests the molecules inhibit RNA splicing, leaving the missing exon-7 intact.

Jones et al. went on to investigate SMN-C3 further; in their study on how SMN-C3 impacted gene transcription, it was observed that aside from no significant change in mRNA abundance for SMN2, 6 genes were upregulated while 6 were downregulated. This poses a potential risk for treatment with SMN-C3, as the change in regulation for functional genes could have unexpected side effects.

Further studies in an in vivo mouse model showed that the oral administration of SMN-C3 resulted in an increase in SMN protein over a period of 10 days, showing it has the intended biological effect. It also suggest that doses should continue to see a long-term effect as once the doses stopped being administered SMN levels dropped. This suggests the protein is unstable.

SMN-C3 has potential to be a treatment option for SMA, though it should be noted that long-term use could have side effects in human trials.

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In this study, 3 compounds SMN-C1, SMN-C2, SMN-C3 were tested against their ability to downregulate the splicing of exon7 (∆7) of the SMN2 gene in an attempt to treat Spinal Muscular Atrophy patients. In the in vitro experiments, the 3 compounds were all observed to upregulate full length of the SMN2 gene and downregulate the prevalence of the spliced ∆7 gene. SMN-C3 in particular was further studied on its effect on the up and downregulation of different genes, with 19 genes upregulated and 25 genes downregulated. In the in vivo study using mice, a daily dose of SMN-C3 was shown to increase the production of the SMN protein, coded from the SMN2 gene necessary to prevent SMA. After 10 days of daily dosage of SMN-C3, the mice were observed to have a much higher amount of SMN protein than their initial Day 1 dosage, but with the amount falling overtime. Fortunately, SMN-C3 could be a potential treatment for SMA. However, a treatment does not mean a cure. SMN-C3 must be taken daily in order to maintain a high level of SMN protein. Another worrisome point is the fact that there are many genes that have either been upregulated or downregulated by SMN-C3, these side effects were not observed, but it does not mean it won’t be present in humans.

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Three compounds: SMN-C1, SMN-C2 and SMN-C3 were used to test if levels of functional SMN2 could be increased pharmacologically, and whether this would be effective in in vivo models. Addition of all three compounds caused increased expression of functional full length SMN2 and reduced expression of SMN2 with missing exon 7, suggesting their potential as a therapeutic drug to increase functional SMN2 levels. Notably, these compounds worked at low concentrations of ≤1 µM, and hence unlikely cytotoxic to cells. Compared to DMSO-treated SMA patient fibroblasts, SMN-C3-treated SMA patient fibroblasts did not show significant changes in SMN2 mRNA abundance. This suggests that the rate of alternative splicing upon addition of SMN-C3 was not altered. However, the effects of SMN-C1 and SMN-C2 on the rates of alternative splicing was not assessed.

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Figure 1 shows that as the concentration of all 3 drugs increased, when applied to the SMA fibroblasts, the relative transcript level of the full length (healthy) mRNA also increased and the relative transcript level of the SMA-causing D7 exon mRNA decreased. This provides a good indication of whether the 3 SMN drugs reduce SMA-causing mRNA. An alternate study was also produced to show whether SMN drugs had no effect on the transcription of other genes. This study is summarised in Figure 2, and it clearly shows that SMN-C3 only affected the total transcript expression of a handful of genes. The figure also shows how the change in transcription level of SMN2 in SMN-C3-treated cells isn’t significant when compared to the DMSO control. The final step would be to test the impact of the 3 drugs in an in vivo subject. Figure 3A shows us how the SMA-positive mice successfully metabolised the SMN-C3 drug as the levels decreased both in the plasma and brain. Figure 3B shows us how the mice responded more strongly to the drug after continuous daily dosage for a prolonged period. The use of these drugs against SMA seems promising and it warrants further research. However, mice are not perfect human models and there could be some hidden risks

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Jones et al. investigated the potential of 3 compounds, SMN-C1, SMN-C2 and SMN-C3 as treatments for SMA. SMA is caused by mutations that trigger alternative splicing of the SMN2 gene; exon-7 is removed, leading to an unstable protein that is degraded.
Figure 1 shows that all 3 compounds caused an increase in the relative transcription level of the healthy SMN2 gene. It also shows that the same 3 compounds simultaneously caused a decrease in the relative transcription level of the alternatively spliced SMN2 gene, stopping SMA characteristics from taking shape.
Figure 2 shows that SMN-C3 treatment does not have a significant impact on the up/down-regulation of the SMN2 gene. It essentially combines the results from Figure 1 and you cannot tell which form of the gene is affected.
Figure 3 A shows that a high concentration of SMN-C3 is present in the brain 3 hours after treatment while a much lower concentration of SMN-C3 is present in the plasma. These values level out after ~24 hours.
Figure 3 B shows that the levels of the SMN protein increased for 24 hours on Day 1 in comparison to 6 hours on Day 10.
I think these results, particularly Figure 1, show that SMN-C3 treatment may be viable since it significantly lowers the transcription of the alternatively spliced (diseased) gene. However, this would need to be tested in vivo in humans to be sure.

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Three compounds, SMN-C1, SMN-C2, and SMN-C3, were applied to fibroblasts derived from SMA patients. All three compounds decreased the alternatively spliced form missing exon (∆7) in a dose-dependent manner. This shows that these three compounds can effectively inhibit the alternative splicing of SMN2 mRNA.

Then, SMN-C3 was applied to patient fibroblasts and the gene transcription was measured. While the transcription level of most genes, including SMN2, did not show a significant change, 6 genes were upregulated, while 6 downregulated.

Next, SMN-C1, C2 and C3 were given to SMA mouse orally. SMN-C3 reached peak concentrations in both the plasma and the brain about 3 hours after administration, while showing a major target site in the brain. SMN-C3 was given to the mouse for 10 days and the level of SMN was measured on Day 1 & 10. 24 hours after the first administration, SMN protein level in quadriceps and brain increased. After 10 days of treatment, the SMN level is 2-2.5 times of that on Day 0, and the level remains higher than Day 0 even 7 days after the treatment stopped.

This study shows the SMN-C1, C2, C3 compounds can inhibit alternative splicing of SMN2 mRNA to generate the mutant ∆7 responsible for SMA, while not significantly changing the mRNA expression of SMN2. In addition, when SMN-C3 was given to SMA mouse, it was shown that it mainly acted in the brain. The effect includes an increased level of SMN protein in quadriceps and the brain.

This study introduced some interesting compounds that can potentially be used in treating SMA, by inhibiting the mutation caused by the alternative splicing of SMN2. In addition, in vivo experiments in mice also showed promising result, with the level of full length SMN protein increasing in quadriceps and the brain.

One concern is that SMN-C3 can lead to the over- or under-expression of some genes, which might have unexpected consequences. These genes need to be identified and the effect of the change of transcription studied. Also, possible side effects should be studied both in mouth and in clinical trials.

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Jones et al. performed a series of experiments on three mice; this involved monitoring the levels of SMN produced in the brain and quadriceps in the hours following oral administration of SMN-C3 at a concentration of 10 mg/kg. The experiments showed that the levels of SMN increased dramatically - up to an 80% increase from baseline 24 hours after administration. The data provided by Jones et al. could offer a potential new treatment for spinal musical atrophy (SMA); the data shows that when treated with SMN-C3, the levels of the mutant variant ∆7 (which is responsible for causing SMA) decrease to 0. This is a significant discovery, treatment of individuals suffering from SMA could be changed drastically, with the real possibility of a better quality of life. However, there are risks; the data shows that SMN2 (which codes for SMN protein), isn’t the only gene to ne affected by the treatment. A total of nearly 40 genes were up or down regulated, which could cause significant changes to the individual being treated.

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Jones et al performed studies trying to discover how the group of SMN compounds affected cells. In one experiment fibroblast cells were exposed to increasing concentrations of SMN C-1, C-2 and C-3. The results showed that as the concentrations of the SMN compounds increased, the number of full-length mRNA transcripts of the SMN2 gene increased while the number of delta 7 transcripts decreased. This indicates that the SMN compounds have real physiological effects. Another experiment implied that although SMN-C3 altered splicing in cells, the total amount of mRNA generated did not change. A third experiment showed that given SMN compounds were found at a greater concentration in neural cells than in the blood. Altogether these experiments show that SMN compounds are taken up by neural cells from the blood, where they influence the splicing of the SMN2 gene which causes more full-length mRNA transcripts to be generated. These transcripts will cause a different proteome than normal to be produced, which could influence cellular behaviour. This data shows that the SMN compounds have a physiological effect upon cells.

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The investigation by Jones et al. lead to the discovery of supporting evidence that treating SMA patient fibroblasts with SMN-C1, -C2 and C3 showed positive results. Experiments where RT-PCR was measured showed that SMN-C1, -C2 and C3 had an effect on RNA splicing. The effect seen was that there was a decrease in the relative transcription level of SMN2 missing exon 7 while there was an increase in relative transcript levels of treated cells, SMN-C1 and -C2 showing the highest levels. Further analyses showed that the treatment of fibroblasts seem to not cause any transcriptional changes in SMN2 seeing as no significant change in total mRNA abundance was detected. In fact, overall expression was maintained seen as very few proteins strayed from normal transcription implying there was no significant impact on transcriptional up or downregulation or splicing. However, experiments measuring SMN proteins in brain and quadricep tissue did show at least an overall 50% increase from baseline of protein expression. This effect was seen both after 1 day and 10 days of treatment implying that the length of treatment has no effect on protein retained by these tissues.

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When fibroblast cells from infected patients were exposed to these new compounds, there was an increase in the full length mRNA and decrease in the faulty mRNA. This appears to have led to increase in the concentration of SMN protein as the mRNA can be transcribed. The findings of Jones et al, suggest a new compound that could potentially be used to treat SMA, however it should be noted that the compound and it effects are not long term and so it’s likely that this medication would have to be taken quite regularly. There is also a significant difference between the concentration of SMN protein produced, although there is an increase in both, so it is possible that repeated exposure to the compounds could cause unwanted side effects. Further research must be done in order to ensure this type of treatment is viable.

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From the experiment, SMN-C1, SMN-C2 and SMN-C3 were shown to increase level of full length SMN2 mRNA when applied to fibroblasts from SMA patients.Scientists may be able to increase functional SMN-2 proteins level to alleviate SMA symptoms by applying these small molecules. Further analysis on SMN-C3 impact on gene transcription reveals that there is no significant change in total SMN2 mRNA abundance. By looking at both of these two observations, it is suggested that the proportion of functional SMN2 mRNA increases with increasing SMN-C3 concentration. As such, the loss of exon 7 is reduced.

Inside SMA mouse models, the daily oral dose of SMN-C3 increased SMN protein in both muscle and brain tissues over time. A greater initial SMN protein level in both quadriceps and brain tissues on day 10 than day 1 after the dose was shown in figure 2B.

Another finding is that both plasma and brain SMN-C3 level seemed to plateau above its initial level 24 hours after the dose. Even though plasma and brain SMN-C3 concentration decreases quickly after the dose, it remains higher than before treatment. Hence, the impact of SMN-C3 is shown to be possibly long-lasting in SMA mice. Since mice are mammals, they are genetically comparable to humans. Scientists may be able to translate this finding to SMN-C3 application to SMA patients.

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Jones et al. investigated the effect of the SMN-C1, SMN-C2, and SMN-C3 compounds on alternative mRNA splicing by measuring the relative transcription level of the full length SMN2 gene, and the alternative spliced form missing exon 7. Studies showed that SMN-C1 and SMN-C2 increased transcription level of the full-length SMN2 gene compared to the internal control, GAPDH. In addition, SMN-C2 and SMN-C3 were shown to have a larger effect on reducing transcription of the alternative spliced form of SMN2. The data also showed more significant effect on the transcription level at higher concentrations of the compound. Investigating SMN-C3 in detail showed that the compound resulted in a more than 2 fold change in the up or down regulation of 14 off-target genes, and the compound was absorbed more in certain areas of the body such as the quadriceps and brain.

The data indicates that SMN-C3 does have a measurable effect on increasing transcription of the full length SMN2 gene and decreasing transcription of the alternatively spliced form missing exon 7, suggesting that it could be a potential target for therapeutic interventions for SMA. However, SMN-C3 showed some off-target effects and had stronger absorbance in some tissues than others. This needs to be carefully studied to determine the other effects that the drug could have.

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Three pharmacological agents SMC-C1, SMN-C2 and CMN-C3 were investigated. Levels of mRNA transcripts coding for SMN2 protein were quantified before and after treatment with each molecule, and in each case, the relative levels of normal functioning SMN2 transcripts increased after treatment. This finding was complemented by observed decreasing levels of the abnormal, mutated mRNA transcript after each treatment. These observations provide encouraging initial evidence that levels of SMN2 could be increased pharmacologically.

The efficacy of each tested compound were then compared. SMN-C3 was tested on an in vivo model (mouse) and SMN2 levels in the brain and quadriceps were shown to initially increase after administering a dose of SMN-C3. However, when measuring the difference in total transcript expression in fibroblast cells after treatment with SMN-C3, only 7 out of 11,714 genes appeared to be upregulated, and a further 7 were downregulated. Importantly, the SMN2 gene was not one of the 7 that were found to be significantly upregulated by the compound. This data shows that although positive changes in SMN2 levels were observed after treatment with SMN-C3, the changes may not be significant enough in comparison with SMN-C1 and SMN-C2.

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The screening brought to light three compounds (SMN-C1, SMN-C2 and SMN-C3) that could potentially increase the level of functional SMN2. These were applied on patient fibroblasts in concentrations from 0.01 to 1 μM. Real Time Polymerase Chain reaction results revealed that all compounds dose-dependently increased the expression of the functional SMN2 gene and decreased the relative transcript level of the SMNΔ7 gene. Interestingly, when cells were treated with 1 μM of SMN-C3, no more SMNΔ7 protein was observed in the cells, which points towards these compounds being effective in vitro. To assess the safety of the most promising compound (SMN-C3), its effect on the transcription levels of all the genes in fibroblasts was conducted, with positive results: only 1% of genes’ transcription was significantly affected and SMN2 was not one of them. This suggests that SMN-C3 does not affect gene expression itself, but the process of splicing the premature mRNA into its mature form, allowing it to contain exon 7. Perhaps even more promising is the performance of SMN-C3 in in-vivo models. When a daily dose was administered to a mouse model, SMN2 levels in both brain and muscle were elevated by 50% on day one and up to 250% on day 10, suggesting the drug becomes more effective in time. These findings can serve as a building block for future research and even clinical trials that could examine the effect of SMN-C3 in humans. However, there are some risks that need to be addressed, mainly that an optimal dose has not been established yet, as well as a lack of understanding about how this drug operates in humans. These questions need to be answered before it can be truly said that there might be a treatment for SMA, but these findings are certainly a step in the right direction.

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During experiments Jones et. al. showed that the three compounds SMN-C1/C2/C3 decreased the level of alternatively spliced RNA and increased full length SMN2. Highlighting a beneficiary effect for Spinal muscle atrophy (SMA) patients, suggesting a possible future treatment.  This is due to the fact there is a higher level of alternatively spliced SMN2 gene in SMA patients causing muscle to be degraded. In mouse models it was shown that in vivo there is an increase in SMN protein in the first 24 hours for the quadriceps and 12 hours in the brain. Following the initial time period there is a gradual decrease in SMN protein levels. This shows that SMN-C1/C2/C3 do not have lasting effects therefore the compound must be taken regularly.

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The three compounds used are SMN-C1, SMN-C2, and SMN-C3. They all showed similar effects in reducing alternative splicing that causes mutated SMN2 with the total transcription level of SMN2 unchanged in fibroblasts. When tested in vivo on mice models, an increase of the compound concentration can be found in the plasma and brain within 6 hours. This contributes to an increase in SMN2 protein levels in the brain and quadriceps. The SMN2 protein can accumulate in the mice as well after doses of SMN-C1, SMN-C2, or SMN-C3 are given and slowly decrease to initial level after 168 hours post-dose, meaning that the compounds can have relatively  long-lasting effect if given regularly.
The findings show a promising application of the three compounds as treatments for SMA by decreasing the mutated alternative splicing and increasing functional SMN2 in vivo. However, the discoveries are yet based on mice model and in vitro studies so further trials need to be taken to prove that they are applicable in humans. Also, although the SMN2 level is not significantly changed by the compounds in patient fibroblasts, it is found that 6 genes can be up-regulated and 6 genes are down-regulated by the treatment of these compounds. This could pose a potential risk to patients and thus further studies on the side effects of the compounds are needed.

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This data shows the effect of the chemicals SMN-C1, SMN-C2 and SMN-C3 on transcription, specifically of the SMN2 gene, which is identical to the SMN1 gene and codes for about 15% of the SMN protein production in the human body.Figure 1 shows that SMN-C1 increases the relative transcriptional level the most, followed by SMN-C2, whilst SMN-C3 isn’t very effective. The effect of SMN-C3 very slightly downregulates the SMN2 gene, but not by a significant level. It upregulates about 6 genes by double and downregulates the same amount by around half. In the third figure, we can see that the SMN-C3 increases the concentration of SMN protein in the quadriceps and brain, peaking at 12-24 hours after the dose and gradually decreasing after that. This suggests repeated doses might be required.The broader significance of this essay is that people with spinal muscular atrophy (SMA) might have a cure. The risks of these compounds is that other genes might be affected by them, which could have unforeseen side effects.

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This data shows the effect of the chemicals SMN-C1, SMN-C2 and SMN-C3 on transcription, specifically of the SMN2 gene, which is identical to the SMN1 gene and codes for about 15% of the SMN protein production in the human body.Figure 1 shows that SMN-C1 increases the relative transcriptional level the most, followed by SMN-C2, whilst SMN-C3 isn’t very effective. The effect of SMN-C3 very slightly downregulates the SMN2 gene, but not by a significant level. It upregulates about 6 genes by double and downregulates the same amount by around half. In the third figure, we can see that the SMN-C3 increases the concentration of SMN protein in the quadriceps and brain, peaking at 12-24 hours after the dose and gradually decreasing after that. This suggests repeated doses might be required.The broader significance of this essay is that people with spinal muscular atrophy (SMA) might have a cure. The risks of these compounds is that other genes might be affected by them, which could have unforeseen side effects.

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Primary results are promising. Especially for SMN-C3 which has been studied in more depth.

In vitro, treatment with increasing SMN-C1, C2 and C3 was correlated with increased transcript of full length SMN2 and a decrease in delta7 SMN, the shortened unfunctional protein.

Data is missing for SMN-C1 and C2 but figure 2 shows that treatment with SMN-C3 does not affect significantly SMN ene expression. Meaning that the increase in the SMN2 protein aimed at would be due to the action of the treatment on the splicing of the mRNA and not gene expression.

Treating living mouse with SMN-C3 led to an increase in brain and muscle SMN2 for the approximate 40 next hours, before the treatment has been metabolized and the SMN levels go back to their original point.

This treatment seems to be promising as it is a simple oral administration. However, data is missing to assess the efficiency of SMN-C1 and C2. Also, the time lapse of SMN-C3 efficiency means that the treatment should be administered every day or so. Finally, further research should be done to assess any potential secondary effects and changes in effects from mouse to humans.

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The effect of 3 different drugs, SMN-C1, SMN-C2, and SMN-C3, were applied to fibroblasts from SMA patients to see its effects on the total transcription the SMN2 gene and the alternatively spliced form that is missing exon 7. The data suggests that all three drugs increase the transcription of the full-length gene and decreases the form missing exon 7. This indicates that the drug can be effective in increasing the level of functional SMN2. The additional effects on other genes from SMN-C3 was also studied and showed that 7 genes increased in expression by double or more and 7 other genes reduced their expression by half or more. Additionally, SMN2 did not increase in gene expression by a significant amount in comparison to the other genes that have had drastic changes in gene expression when exposed to SMN-C3. Moreover, the drug does increase SMN found in quadriceps and brain tissue however, overtime the amount lowers. The investigation by Jones et al. suggests that dugs can manipulate the expression of functional SMN decreasing the amount of negative alternative splicing. Even so, the drugs still have effects on other genes expressed in fibroblast and requires further testing to see if this may be harmful or creates risk for the individual.

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Jonas et al study essentially focusses on the effects of SMN-C for treatment of SMA. As SMN-C1 concentration increases, the full length of mRNA increases while the spliced mRNA gradually decreases. The overlapping error bars and similar trend lines therefore prove that SMN-C decreases the RNA splicing process.

Jonas et al then investigated the effect of SMN-C3 on gene expression. Figure 2 highlights that SMN-C3 can up and down regulate almost an equal number of genes, however, most of the genes have a Log2 fold change of 0. This suggests that SMNC-3 has no significant change in total mRNA abundance, especially for SMN2.

SMN-C3 is in higher concentrations in the brain compared to the plasma, and peaks before the first 6 hours. Continuous treatment is required to increase the SMN2 baseline, as after one day there is small peak which quickly decrease, but after 10 continuous days of treatment, there is a much larger peak and the protein decreases comparatively more gradually.

Overall, this study is significant as we found that SMN-C3 will help treat SMA as increases healthy SMn2 gene expression. Yet, it is important to investigate the significance of this treatment and its side effects, especially after daily dosage.

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Jones et al.’s study shows that SMN-C1, SMN-C2 and SMN-C3 compounds have the ability to reduce the level of gene expression of spliced forms of mutated SMN, which are missing exon 7.

To understand how SMN-C1, C2, and C3 molecules prevent the ∆7 form of alternative splicing, an experiment of mRNA SMN abundance was conducted in fibroblasts treated with SMN-C3. The results showed that SMN-C3 does affect alternative splicing in fibroblasts. However, changes in SMN gene expression is not the reason why. Therefore, it can be assumed that SMN-C3, and the others, alter splicing during the mRNA maturation process. Further experiments on mice indicated that the drug has a higher effect the longer it has been used. The starting level of SMN on the first day of SMN-C1, C2 and C3 oral dosage was 0, which was increased to 100% change from baseline on day 10.

This study has proven that SMN-C1, C2, and C3 are effective in reducing the levels of transcription of mutant SMN mRNA, especially SMN-C1 and SMN-C2. However, as the in vivo was performed on mice, before using it as treatment it would have to go through a clinical trial.

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The in vivo study by Jones et al has indicated that SMN-C1, C2, and C3 can increase the levels of healthy SMN protein compared to baseline of SMA mouse on both days 1 and 10. There is a larger increase in healthy SMN protein after drug intake on day 10 compared to day 1, potentially suggesting increased drug effectiveness over time. There is also a larger starting amount of health SMN protein (130% compared to baseline) on day 10 compared to day 1 (0%). Given these result, there could be potential applications for treating SMA in humans if the drug is given on a daily or bidaily basis given that further trials are done in humans to confirm that it is safe for humans. There is potential risks when transferring from a mouse in vivo model compared to humans as the metabolism (ingested orally) of the drugs could be very different and have unwanted side effects in humans that is not observed in mice.

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Jones et al chose SMN C1, SMN-C2 and SMN-C3 as the compounds to add to the fibroblasts. Through the experimentation he provided a link between and increase in the concentration of all of these compounds and an increase in the full length of the SMA mRNA and, consequentially, and increase in the mRNA containing the un-spliced crucial exon 7. This link is further proved by another in vivo experiment that shows SMN protein levels decreasing post-treatment with these compounds. Furthermore, Jones et al showed that mRNA abundance remained unaltered after treatment with SMN-C3 treatment which potentially indicates that these compounds do not change the amount of mRNA expressed but rather promote the presence of exon 7 which explains the increase in full length transcription of mRNA and the decrease of SMN functional protein following the use of this compounds.

This study could have many potential applications and shows promise as a ‘magical bullet’ for this non-infectious disease. Further studies could be used to determine the best time in the infant development for these drugs should be introduced and where- in the germ line or the somatic cell line.

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Jones et al. decided to test the effects of SMN-C1, SMN-C2 and SMN-C3 as possible treatments to SMA, by identifying the changes in expression of the different spliced SMN2 gene. They showed that increasing the concentrations of the compounds raised the concentration of the healthy full-length gene and reduced the concentration of the mRNA that was missing exon 7.  Furthermore, they presented in figure 2 that treatment with SMN-C3 did influence the expression of some genes, however the change in expression of SMN2 wasn’t significant. After this they continued their experiments in vivo, using mice to as their model to show the effects. The data in figure 3 reveals that the compound SMN-C3 tends to accumulate in the brain significantly more than it does in the plasma, with a peak in concentration at after only a few hours. It also reveals that continuous treatment is required to significantly increase the SMN2 expression above baseline. This is because after only 1 day of treatment, the expression only lasts a few hours before it reaches a small peak and quickly decreases. However, after 10 days of continuous treatment, it has a far larger peak and takes approximately 7 days to return to baseline. This data tends to show that treatment with SMN-C compounds increase the expression of the healthy SMN2 gene expression, possibly presenting itself as a treatment for SMA. However, many more questions need to be asked, such as the effect of expression shown in figure 2 and if these side effects outweigh the effects of SMA and its currently available treatments.

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In a series of experiment, Jones et al. were able to show that by giving molecules of SMN-C1, C2, and C3 it not only increased the level of transcript of the functioning full length RNA transcript as opposed to the missing exon 7 transcript, but also that the molecules were taken in on the in vivo model and correlated with SMN protein level in both the quadriceps and the brain. Furthermore, it shows that these molecules were able to interfere with alternate splicing and produce functional RNA transcripts of the defective gene. This give insights into potential treatment options for patients with SMA, however it seems that the effects of the molecules seem to be temporary and needs further doses to sustain the effects. Therefore, these molecules need to be dealt with serious and proper testing to ensure it’s safety for long-term use, which means looking at what other genes it is affecting and whether it involves oncogenes or have adverse effects to other genes it might involve.

# data[].attributes.headline
The study tested three compounds, SMN-C1, SMN-C2, and SMN-C3, both on fibroblasts derived from SMA patients and on mice fibroblasts. The aim was to investigate whether these compounds could reduce the SMN2 mRNAs lacking exon 7 and instead produce full length SMN2 mRNAs which would translate into a functional protein. The findings were very encouraging. All three compounds showed a decrease in SMN2 transcript lacking exon 7 and an increase in the full length SMN2 transcript. Additionally, there wasn’t a significant difference in the effectiveness of the three compounds. An important risk to consider is that these compounds could lead to an overall increase in transcription of SMN2 which would surpass its normal levels. However, the second experiment demonstrated that although SMN2 was being transcribed in full length, its relative abundance hadn’t significantly changed. Lastly, the three compounds had to be tested on an organism to test their effects in vivo. Mice models of SMA were used for this investigation. First finding was that within the 24 hours of administration, SMN-C3 could be found in greater quantities in the brain than in the plasma. Additionally, the SMN protein levels in the brain and quadriceps after 10 days of SMN-C3 administration were decreased in comparison to baseline levels. This raises important questions. 

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The study tested three compounds, SMN-C1, SMN-C2, and SMN-C3, both on fibroblasts derived from SMA patients and on mice fibroblasts. The aim was to investigate whether these compounds could reduce the SMN2 mRNAs lacking exon 7 and instead produce full length SMN2 mRNAs which would translate into a functional protein. The findings were very encouraging. All three compounds showed a decrease in SMN2 transcript lacking exon 7 and an increase in the full length SMN2 transcript. Additionally, there wasn’t a significant difference in the effectiveness of the three compounds. An important risk to consider is that these compounds could lead to an overall increase in transcription of SMN2 which would surpass its normal levels. However, the second experiment demonstrated that although SMN2 was being transcribed in full length, its relative abundance hadn’t significantly changed. Lastly, the three compounds had to be tested on an organism to test their effects in vivo. Mice models of SMA were used for this investigation. First finding was that within the 24 hours of administration, SMN-C3 could be found in greater quantities in the brain than in the plasma. Additionally, the SMN protein levels in the brain and quadriceps after 10 days of SMN-C3 administration were decreased in comparison to baseline levels. This raises important questions. 

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The study looked at molecules SMN-C1, C2 and C3 and their effects on alternative splicing. Results showed that application of either of the three compounds led to a substantial decrease in the spliced RNA missing exon 7 with an increase in full-length RNA. It is likely that a reduction of the spliced form of RNA missing exon 7 would lead to increased survival of SMN2. The in vivo experiments showed that upon administration, there was a spike in the concentration of the molecule administered in both plasma and the brain, followed by an increase in the total concentration change in SMN protein concentration in mice quadriceps and the brain. The drug was administered to mice on days 1 and 10. What is particularly interesting, though, is that on day 10, the initial percent change of SMN2 concentration was already higher than that on day 10, and as time passed there was a sharp increase. This proposes that if the drug is taken continuously, the baseline concentration of functional SMN2 could be increased, thus providing a long-term therapeutic option.

Although Jones et al. also showed that SMN2 has no significant change in mRNA abundance, the abundance of 6 genes doubled and halved for 6 others. This begs some questions regarding the risks. What are these 12 genes, and does their mRNA abundance have any adverse side effects? What happens when the quantity of these genes is doubled? Halved? These are crucial points to consider before moving forward with studies on SMN-C1, C2 and C3. Nonetheless, Jones et al. showed optimistic findings regarding the possibility of increasing the level of SMN2 pharmacologically, which could ultimately prove to be significant in future research on SMA treatments and therapeutics.

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Three different compounds were tested as possible pharmacological treatments: SMN-C1, SMN-C2 and SMN-C3. These were applied to the fibroblasts of SMA patients and PCR was used to quantify the SMN2 gene and the exon 7.  All three compounds have increased the transcriptional level of the SCM2. Additionally, all have decreased the levels of exon 7, meaning that it has stayed as a part of the SCM2 and haven’t been spliced out.

The abundance of the SMN2 gene is increased by 1.5 times with the addition of SCM-3 compound compared to a fibroblast of a SMA patient and in vivo models the SCM-C3 has also shown to increase the amounts of SCM-C1 produced naturally by the mice. The mice have also produced a significantly increased amounts of the SMN

protein in both the quadriceps and the brain, but it seems like the effect of the SCM-C3 has decreased over time.

All of the above means that SCM-C3 increases the production of naturally occurring SCM-C1 which in turn upregulates the transcription of the full SCM gene that includes the exon 7 to produce a functioning protein. Therefore, this compound could potentially help the individuals with spinal muscular dystrophy as it helps them to produce a functional protein.