Research Round-Up

Posted on at February 22, 2018
9011 1,995

In neurons…

The HD Induced Pluripotent Stem Cell (iPSC) consortium differentiated HD patient iPSCs into neuronal cultures.1 RNA-seq and ChIPseq analysis from these cultures indicated subtle alterations in expression and epigenetic signature of genes involved in neural development, function, and striatal maturation in the presence of mHTT. The small molecule isoxazole-9 is known to target some of the disrupted gene networks, and when tested in this system, was able to normalize some of the genetic phenotypes. This compound also showed beneficial effects in HD-related pathologies in the R6/2 mouse model, indicating that some of these pathways are linked to pathology. Yu and Tanese used mouse embryonic stem cells provided by the Zeitland group to investigate HTT involvement in differentiation.2 The lines they used came from the same background, but were either huntingtin-null (Htt-null) or had tagged wildtype Htt (Q7) or mHtt (Q140).

The group found that the presence of HTT, either wildtype or mutant, allowed all three germ layers to form, but Htt knockout blocked the ability of the cells to differentiate down the ectoderm pathway to form neural stem cells (NSC). Although mHTT still allowed NSC formation, it inhibited further differentiation into neurons. RNA-seq data from these lines revealed that MaoA and Oligo1/2 downregulation and upregulation of the PRC2 repressor complex may be involved in these developmental changes.

One well-described effect of mHTT is the dysregulation of Ca2+ signaling. A new drug lead that corrects deficiencies in storeoperated calcium entry (SOCE) in YAC128 medium spiny neurons (MSNs) has been described in a recent publication.3 SOCE was found to be increased specifically in MSNs but not in other neuronal subtypes in this model.

The group identified a tetrahydrocarbazole that attenuated this phenotype in addition to normalizing mitochondrial function. The group postulates that the dual benefit might come from preventing abnormal Ca2+ accumulation in the mitochondria and endoplasmic reticulum.

1HD iPSC Consortium. Developmental alterations in Huntington’s disease neural cells and pharmacological rescue in cells and mice. Nat Neurosci. 2017 Mar 20. doi: 10.1038/nn.4532. [Epub ahead of print].

2Yu MS, Tanese N. Huntingtin Is Required for Neural But Not Cardiac/Pancreatic Progenitor Differentiation of Mouse Embryonic Stem Cells In vitro. Front Cell Neurosci. 2017;11(33).

3Czeredys M, Maciag F, Methner A, Kuznicki J. Tetrahydrocarbazoles decrease elevated SOCE in medium spiny neurons from transgenic YAC128 mice, a model of Huntington’s disease. Biochem Biophys Res Commun. 2017 Feb 19;483(4):1194-1205. 23;116:233-246.

In gene editing…

Allele-specific silencing of mHTT with methods that degrade mHTT mRNA is a promising treatment for HD, but that silencing is neither complete nor permanent. The discovery of the CRISPR/Cas9 nuclease system has created the possibility of altering genomic DNA in affected cells, and permanently silencing mHTT.

Two groups have recently published studies that attempt this in mice.1,2 Both groups utilized the existence of allele-specific single-nucleotide polymorphisms (SNPs) that create or destroy protospacer adjacent motif (PAM) elements in an allele-specific manner in the mHtt promoter. The groups then used a two-guide RNA (gRNA) system, with one gRNA targeted against the SNP in the promoter, and one in a downstream intron, leading to excision of the mHtt promoter and first few exons.

The Davidson group used this technique, excising exon1 of mHtt in an allele-specific manner, in immortalized cells, in human HD fibroblasts, and in vivo in a BACHD mouse model.1 Shin and colleagues excised a 44 kb segment of mHtt, including exons 1–3, and successfully used their platform in induced pluripotent stem cells and neural precursor cells.

2 Together, these data demonstrate the reproducibility and utility of using the CRISPR nuclease system to potentially permanently alter DNA to silence mHTT for therapeutic effect. CRISPR/Cas9 also can be used to create tools for drug screening. The Pouladi group coupled CRISPR/Cas9 technology with piggyback transposon technology to create corrected isogenic HD lines.3 The corrected lines maintained their pluripotency, differentiation potential, and karyotype, with no off-target effects.

The correction restored HD-induced phenotypes, including neural rosette formation deficiencies, apoptotic susceptibility, and energetic defects in the isogenic iPSC line. Performing global differential gene expression analysis on the parental line, the corrected line, and a non-related control line allowed the scientists to determine which genes were differentially expressed because of the HD mutation, and which genes were differentially expressed because of genetic background.

1Monteys AM, Ebanks SA, Keiser MS, Davidson BL. CRISPR/Cas9 Editing of the Mutant Huntingtin Allele In Vitro and In Vivo. Mol Ther. 2017 Jan 4;25(1):12-23. doi: 10.1016/j.ymthe.2016.11.010. Epub 2017 Jan 4. 2Shin JW, Kim K-H, Chao MJ, et al. Permanent inactivation of Huntington’s disease mutation by personalized allele-specific CRISPR/ Cas9. Hum Mol Genet. 2016 Oct 15;25(20):4566-4576. 3Xu X, Tay Y, Sim B, et al. Reversal of Phenotypic Abnormalities by CRISPR/Cas9-Mediated Gene Correction in Huntington Disease Patient- Derived Induced Pluripotent Stem Cells. Stem Cell Reports.2017 Mar 14;8(3):619-633.

In imaging…

Dr. Sarah Tabrizi’s group continues to datamine the extensive human imaging and disease phenotype data available from the two large multi center cohort studies, Track-HD and Track-On HD. In the largest imaging study ever done to assess depressive symptoms in HD, the group examined variation in structural and functional brain networks in relation to symptoms of depression in premanifest HD (preHD) and healthy controls.1 Onset of depression precedes onset of motor symptoms, and neuroimaging studies may reveal the early affected brain networks. The group found that depressive symptoms in preHD are correlated with specific connectivity changes in the default mode network and the basal ganglia.

Another group published its imaging and electrophysiological study in Nature’s Scientific Reports, looking for functional and structural changes that are present prior to clinical diagnosis.2 This study looked at preHD patients who had a very low disease burden score, far from disease onset. All methods used in this study agree that brain function and structure is normal in this cohort, meaning that symptoms are reflective of an ongoing neurodegenerative process. This may have implications for timing of treatment. Finally, another large-scale patient-based collaborative effort aimed to identify patients with extreme HD motor phenotypes with respect to their CAG repeat length and age, in order to search for disease modifiers.3 By looking at more than 10,000 HD patients, the group found that the total motor score (TMS) could vary by up to 30 years in individuals with the same CAG repeat length. They found that a TMS score of more than 13 would predict motor onset regardless of age and CAG length. The identified patients in the upper and lower quantiles can be further examined for biological, medical, or environmental factors that may lead to their extreme motor phenotype, which will inform patient care and prognosis.

1McColgan P, Razi A, Gregory S, et al. Structural and functional brain network correlates of depressive symptoms in premanifest Huntington’s disease. Hum Brain Mapp. 2017 Mar 15. doi: 10.1002/ hbm.23527. [Epub ahead of print]

2Gorges M, Müller H-P, Mayer IMS, et al. Intact sensory-motor network structure and function in far from onset premanifest Huntington’s disease. Sci Rep. 2017 Mar 7;7:43841. 3Braisch U, Hay B, Muche R, et al. Identification of extreme motor phenotypes in Huntington’s disease. Am J Med Genet B Neuropsychiatr Genet. 2017 Apr;174(3):283-294.

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    I want to pass along some very important news that everyone needs to hear!

    In December of 2017, Donald Trump made history by recognizing Jerusalem as the capital of Israel. Why is this big news? Because by this the Jewish people of Israel are now able to press forward in bringing about the Third Temple prophesied in the Bible.

    Jewish Rabbis have publicly announced that their Messiah will be revealed in the coming years who will be a leader and spiritual guide to all nations, gathering all religions under the worship of one God.

    Biblical prophecy tells us that this Jewish Messiah who will take the stage will be the antichrist “who opposes and exalts himself above all that is called God or that is worshiped, so that he sits as God in the temple of God, showing himself that he is God” (2 Thessalonians 2:4). For a time he will bring about a false peace, but “Therefore when you see the ‘abomination of desolation,’ spoken of by Daniel the prophet, standing in the holy place (Matthew 24:15)…then there will be great tribulation, such as has not been since the beginning of the world until this time, no, nor ever shall be” (Matthew 24:21).

    More importantly, the power that runs the world wants to put a RFID microchip in our body making us total slaves to them. This chip matches perfectly with the Mark of the Beast in the Bible, more specifically in Revelation 13:16-18:

    “He causes all, both small and great, rich and poor, free and slave, to receive a mark on their right hand or on their foreheads, and that no one may buy or sell except one who has the mark or the name of the beast, or the number of his name.

    Here is wisdom. Let him who has understanding calculate the number of the beast, for it is the number of a man: His number is 666.”

    Referring to the last days, this could only be speaking of a cashless society, which we have yet to see, but are heading towards. Otherwise, we could still buy or sell without the mark amongst others if physical money was still currency. This Mark couldn’t be spiritual because the word references two different physical locations. If it was spiritual it would just say in the forehead. RFID microchip implant technology will be the future of a one world cashless society containing digital currency. It will be implanted in the right-hand or the forehead, and we cannot buy or sell without it. Revelation 13:11-18 tells us that a false prophet will arise on the world scene doing miracles before men, deceiving them to receive this Mark. Do not be deceived! We must grow strong in Jesus. AT ALL COSTS, DO NOT TAKE IT!

    “Then a third angel followed them, saying with a loud voice, “If anyone worships the beast and his image, and receives his mark on his forehead or on his hand, he himself shall also drink of the wine of the wrath of God, which is poured out full strength into the cup of His indignation. He shall be tormented with fire and brimstone in the presence of the holy angels and in the presence of the Lamb. And the smoke of their torment ascends forever and ever; and they have no rest day or night, who worship the beast and his image, and whoever receives the mark of his name” (Revelation 14:9-11).

    People have been saying the end is coming for many years, but we needed two key things. One, the Third Temple, and two, the technology for a cashless society to fulfill the prophecy of the Mark of the Beast.

    Visit http://WWW.BIBLEFREEDOM.COM to see proof for these things and why the Bible truly is the word of God!

    If you haven’t already, it is time to seek God with all your heart. Jesus loves you more than you could imagine. He wants to have a relationship with you and redeem you from your sins. Turn to Him and repent while there is still hope! This is forever…God bless!

    “EITHER HUMAN INTELLIGENCE ULTIMATELY OWES ITS ORIGIN TO MINDLESS MATTER OR THERE IS A CREATOR…” – JOHN LENNOX

    We all know God exists. Why? Because without Him, we couldn’t prove anything at all. Do we live our lives as if we cannot know anything? No. So why is God necessary? In order to know anything for certain, you would have to know everything, or have revelation from somebody who does. Who is capable of knowing everything? God. So to know anything, you would have to be God, or know God.

    A worldview without God cannot account for the uniformity and intelligibility of nature. And why is it that we can even reason that God is the best explanation for this if there is no God? We are given reason to know or reject God, but never to know that He does not exist.

    It has been calculated by Roger Penrose that the odds of the initial conditions for the big bang to produce the universe that we see to be a number so big, that we could put a zero on every particle in the universe, and even that would not be enough to use every zero. What are the odds that God created the universe? Odds are no such thing. Who of you would gamble your life on one coin flip?

    Is there evidence that the Bible is the truth? Yes. Did you know that the creation accounts listed in the book of Genesis are not only all correct, but are also in the correct chronological order? That the Bible doesn’t say the Earth was formed in six 24-hour days but rather six long but finite periods of time? That the Bible makes 10 times more creation claims than all major “holy” books combined with no contradictions, while these other books have errors in them? The Bible stood alone by concurring with the big bang saying, “In the beginning God created the heaven and the earth” (Genesis 1:1); and says our universe is expanding, thousands of years before scientists discovered these things. Watch a potential life-changing video on the website listed below with Astronomer(PhD) Hugh Ross explaining all these facts based on published scientific data. He has authored many books, backed even by atheist scientists.

    Jesus came to pay a debt that we could not; to be our legal justifier to reconcile us back to a Holy God; only if we are willing to receive Him: “For the wages of sin is death…” (Romans 6:23).

    God so loved the world that He gave us His only begotten son, so that whoever believes in Him, through faith, shall not perish, but have everlasting life. Jesus says if we wish to enter into life to keep the commands! The two greatest commands are to love God with all your heart, soul, strength, and mind; and your neighbor as yourself. All the law hang on these commands. We must be born of and lead by the Holy Spirit, to be called children of God, to inherit the kingdom. If we are willing to humble ourselves in prayer to Jesus, to confess and forsake our sins, He is willing to give the Holy Spirit to those who keep asking of Him; giving us a new heart, leading us into all truth!

    Jesus came to free us from the bondage of sin. The everlasting fire was prepared for the devil and his angels due to disobedience to God’s law. If we do the same, what makes us any different than the devil? Jesus says unless we repent, we shall perish. For sin is the transgression of the law. We must walk in the Spirit so we may not fulfill the lusts of the flesh, being hatred, fornication, drunkenness and the like. Whoever practices such things will not inherit the kingdom (Galatians 5:16-26). If we sin, we may come before Jesus to ask for forgiveness (1 John 2:1-2). Evil thoughts are not sins, but rather temptations. It is not until these thoughts conceive and give birth by our own desires that they become sin (James 1:12-15). When we sin, we become in the likeness of the devil’s image, for he who sins is of the devil (1 John 3:8); but if we obey Jesus, in the image of God. For without holiness, we shall not see the Lord (Hebrews 12:14).

    The oldest religion in the world is holiness through faith (James 1:27). What religion did Adam and Eve follow before the fall? Jesus, Who became the last Adam, what religion does He follow? Is He not holy? He never told us to follow the rituals and traditions of man but to take up our cross and follow Him (Luke 9:23). There are many false doctrines being taught leading people astray. This is why we need the Holy Spirit for discernment. Unlike religion, holiness cannot be created. It is given to us from above by the baptism of the Spirit. Jesus is more than a religion; He is about having a personal relationship with the Father. Start by reading the Gospel of Matthew, to hear the words of God, to know His character and commandments. Follow and obey Jesus, for He is the way, the truth, and the life!

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  380. Hey, how’s it going?

    I want to pass along some very important news that everyone needs to hear!

    In December of 2017, Donald Trump made history by recognizing Jerusalem as the capital of Israel. Why is this big news? Because by this the Jewish people of Israel are now able to press forward in bringing about the Third Temple prophesied in the Bible.

    Jewish Rabbis have publicly announced that their Messiah will be revealed in the coming years who will be a leader and spiritual guide to all nations, gathering all religions under the worship of one God.

    Biblical prophecy tells us that this Jewish Messiah who will take the stage will be the antichrist “who opposes and exalts himself above all that is called God or that is worshiped, so that he sits as God in the temple of God, showing himself that he is God” (2 Thessalonians 2:4). For a time he will bring about a false peace, but “Therefore when you see the ‘abomination of desolation,’ spoken of by Daniel the prophet, standing in the holy place (Matthew 24:15)…then there will be great tribulation, such as has not been since the beginning of the world until this time, no, nor ever shall be” (Matthew 24:21).

    More importantly, the power that runs the world wants to put a RFID microchip in our body making us total slaves to them. This chip matches perfectly with the Mark of the Beast in the Bible, more specifically in Revelation 13:16-18:

    “He causes all, both small and great, rich and poor, free and slave, to receive a mark on their right hand or on their foreheads, and that no one may buy or sell except one who has the mark or the name of the beast, or the number of his name.

    Here is wisdom. Let him who has understanding calculate the number of the beast, for it is the number of a man: His number is 666.”

    Referring to the last days, this could only be speaking of a cashless society, which we have yet to see, but are heading towards. Otherwise, we could still buy or sell without the mark amongst others if physical money was still currency. This Mark couldn’t be spiritual because the word references two different physical locations. If it was spiritual it would just say in the forehead. RFID microchip implant technology will be the future of a one world cashless society containing digital currency. It will be implanted in the right-hand or the forehead, and we cannot buy or sell without it. Revelation 13:11-18 tells us that a false prophet will arise on the world scene doing miracles before men, deceiving them to receive this Mark. Do not be deceived! We must grow strong in Jesus. AT ALL COSTS, DO NOT TAKE IT!

    “Then a third angel followed them, saying with a loud voice, “If anyone worships the beast and his image, and receives his mark on his forehead or on his hand, he himself shall also drink of the wine of the wrath of God, which is poured out full strength into the cup of His indignation. He shall be tormented with fire and brimstone in the presence of the holy angels and in the presence of the Lamb. And the smoke of their torment ascends forever and ever; and they have no rest day or night, who worship the beast and his image, and whoever receives the mark of his name” (Revelation 14:9-11).

    People have been saying the end is coming for many years, but we needed two key things. One, the Third Temple, and two, the technology for a cashless society to fulfill the prophecy of the Mark of the Beast.

    Visit http://WWW.BIBLEFREEDOM.COM to see proof for these things and why the Bible truly is the word of God!

    If you haven’t already, it is time to seek God with all your heart. Jesus loves you more than you could imagine. He wants to have a relationship with you and redeem you from your sins. Turn to Him and repent while there is still hope! This is forever…God bless!

    “EITHER HUMAN INTELLIGENCE ULTIMATELY OWES ITS ORIGIN TO MINDLESS MATTER OR THERE IS A CREATOR…” – JOHN LENNOX

    We all know God exists. Why? Because without Him, we couldn’t prove anything at all. Do we live our lives as if we cannot know anything? No. So why is God necessary? In order to know anything for certain, you would have to know everything, or have revelation from somebody who does. Who is capable of knowing everything? God. So to know anything, you would have to be God, or know God.

    A worldview without God cannot account for the uniformity and intelligibility of nature. And why is it that we can even reason that God is the best explanation for this if there is no God? We are given reason to know or reject God, but never to know that He does not exist.

    It has been calculated by Roger Penrose that the odds of the initial conditions for the big bang to produce the universe that we see to be a number so big, that we could put a zero on every particle in the universe, and even that would not be enough to use every zero. What are the odds that God created the universe? Odds are no such thing. Who of you would gamble your life on one coin flip?

    Is there evidence that the Bible is the truth? Yes. Did you know that the creation accounts listed in the book of Genesis are not only all correct, but are also in the correct chronological order? That the Bible doesn’t say the Earth was formed in six 24-hour days but rather six long but finite periods of time? That the Bible makes 10 times more creation claims than all major “holy” books combined with no contradictions, while these other books have errors in them? The Bible stood alone by concurring with the big bang saying, “In the beginning God created the heaven and the earth” (Genesis 1:1); and says our universe is expanding, thousands of years before scientists discovered these things. Watch a potential life-changing video on the website listed below with Astronomer(PhD) Hugh Ross explaining all these facts based on published scientific data. He has authored many books, backed even by atheist scientists.

    Jesus came to pay a debt that we could not; to be our legal justifier to reconcile us back to a Holy God; only if we are willing to receive Him: “For the wages of sin is death…” (Romans 6:23).

    God so loved the world that He gave us His only begotten son, so that whoever believes in Him, through faith, shall not perish, but have everlasting life. Jesus says if we wish to enter into life to keep the commands! The two greatest commands are to love God with all your heart, soul, strength, and mind; and your neighbor as yourself. All the law hang on these commands. We must be born of and lead by the Holy Spirit, to be called children of God, to inherit the kingdom. If we are willing to humble ourselves in prayer to Jesus, to confess and forsake our sins, He is willing to give the Holy Spirit to those who keep asking of Him; giving us a new heart, leading us into all truth!

    Jesus came to free us from the bondage of sin. The everlasting fire was prepared for the devil and his angels due to disobedience to God’s law. If we do the same, what makes us any different than the devil? Jesus says unless we repent, we shall perish. For sin is the transgression of the law. We must walk in the Spirit so we may not fulfill the lusts of the flesh, being hatred, fornication, drunkenness and the like. Whoever practices such things will not inherit the kingdom (Galatians 5:16-26). If we sin, we may come before Jesus to ask for forgiveness (1 John 2:1-2). Evil thoughts are not sins, but rather temptations. It is not until these thoughts conceive and give birth by our own desires that they become sin (James 1:12-15). When we sin, we become in the likeness of the devil’s image, for he who sins is of the devil (1 John 3:8); but if we obey Jesus, in the image of God. For without holiness, we shall not see the Lord (Hebrews 12:14).

    The oldest religion in the world is holiness through faith (James 1:27). What religion did Adam and Eve follow before the fall? Jesus, Who became the last Adam, what religion does He follow? Is He not holy? He never told us to follow the rituals and traditions of man but to take up our cross and follow Him (Luke 9:23). There are many false doctrines being taught leading people astray. This is why we need the Holy Spirit for discernment. Unlike religion, holiness cannot be created. It is given to us from above by the baptism of the Spirit. Jesus is more than a religion; He is about having a personal relationship with the Father. Start by reading the Gospel of Matthew, to hear the words of God, to know His character and commandments. Follow and obey Jesus, for He is the way, the truth, and the life!

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