Showing posts with label Gene therapy. Show all posts
Showing posts with label Gene therapy. Show all posts

Wednesday 8 May 2024

Immunotherapy from the desert


Today’s post revisits the idea of using immunotherapies to treat autism.

Some readers of this blog are already doing this and a significant percentage of those are using IVIG.

Intravenous immunoglobulin (IVIG) is a pooled antibody, and a biological agent used to manage various immunodeficiency states and a plethora of other conditions, including autoimmune, infectious, and inflammatory states.

IVIG is not a precision therapy, it is more a case of when all else fails try IVIG.

In the United States it seems that many insurance companies will cover the cost of long-term IVIG therapy. In other countries the cost greatly limits the use of this therapy.

An interesting observation is that IVIG products can vary significantly in their potency, depending on where they are made. Several readers of this blog have noted this.

I attended the Autism Challenges and Solutions conference recently in Abu Dhabi. I did have a chat with Laila Alayadhi, a researcher and clinician from Saudi Arabia who has been publishing papers about autoimmunity in ASD for decades. She also published a series of studies that examined the potential of camel milk as a therapy. She examined both changes in biological markers of oxidative stress and inflammation as well as measures of autism severity.

Her most recent study is here:-


Comparative Study on the Ameliorating Effects of Camel Milkas a Dairy Product on Inflammatory Response in Autism Spectrum Disorders

The link between nutrition and autism spectrum disorder (ASD), as a neurodevelopmental disorder exhibiting impaired social interaction, repetitive behavior, and poor communication skills, has provided a hot point of research that might help use nutritional intervention strategies for managing ASD symptoms. This study examined the possible therapeutic potency of raw and boiled camel milk in reducing neuroinflammation in relation to behavioral characteristics. A blinded study was conducted on 64 children with autism (aged 2–12 years). Group I (n = 23) consisted of children who received raw camel milk; Group II (n = 27) comprised children who received boiled camel milk; and Group III (n = 14) comprised children who received cow milk as a placebo. Changes in plasma tumor necrosis factor-alpha (TNF-α) as pro-inflammatory cytokine in relation to behavioral characteristics evaluated using the Childhood Autism Rating Scale (CARS), Social Responsiveness Scale (SRS), and gastrointestinal (GI) symptoms before and after 2 weeks of raw and boiled camel milk therapy. Significantly lower plasma levels of TNF-α were recorded after 2 weeks of camel milk consumption, accompanied by insignificant changes in CARS and significant improvements in SRS and GI symptoms. Alternatively, Group III demonstrated an insignificant TNF-α increase without changes in CARS, SRS, and GI symptoms. This study demonstrated the positive effects of both raw and boiled camel milk in reducing neuroinflammation in patients with ASD. The improvements in the SRS scores and GI symptoms are encouraging. Further trials exploring the potential benefits of camel milk consumption in patients with ASD are highly recommended.



Apparently camel milk tastes just fine, although Dr Alayadhi told us she had never tried it prior to her research. She has shown than both pasteurized and raw milk are equally effective. I did ask her about other types of milk like goat’s milk and she said they had tried other milks and that only camel milk has shown the immunomodulatory effect.  When asked how much you need to drink, the answer was three glasses a day.

The Dentist

I did chat to another Saudi professor, a pediatric dentist, who gave a presentation about treating children with ASD.  Having had some pretty bad experiences with getting dental treatment and then overcoming them, I did feel I had something in common with Ebtissam Murshid.  I did catch up with her later and shared details of the D-Termined program created by US dentist David Tesini. It is a video training program for dentists how to treat kids with autism. I have written about it previously in this blog. Tesini very much tries to make the visit to the dentist fun, with lots of distractions in his treatment room. Murshid purposefully has blank white walls, believing that autistic kids get upset by bright colors and patterns. Hopefully she watches Tesini’s videos.

Murshid has published a book to help parents prepare their children for their trip to the dentist and, like Tesini, had made a small trial to show that her method is effective.

Some dentists are naturally good at treating the most difficult kids, but most are not.  It is impossible to predict.

A really good dentist needs neither restraint, like a papoose board, or sedation. If general anesthetic is needed, then something is not being done right. Kids with severe autism can be treated with local anesthetic just like other kids, they just need to go through a familiarization training like Tesini/Murshid use.


Back to immunotherapy

I did have many conversations with Carmello Rizzo who is an Italian doctor interested in both diet and autoimmunity to treat autism. He is a feature at many autism conferences and is a great speaker. He was telling me about Enzyme Potentiated Desensitization (EPD), an overlooked way to treat allergy care.

EPD was invented in the 1960s by a British immunologist Dr Len McEwen, at St. Mary’s Hospital, Paddington. EPD is approved in the United Kingdom for the treatment of hay fever, food allergy and intolerance and environmental allergies.

It is an unlicensed product (i.e. not a drug), it is available only on a “named patient” basis.

EPD is not the same as allergy shots.

Allergy shots, also known as allergy immunotherapy, are injections used to treat allergies over a long period of time. They work by gradually desensitizing your body to the allergens that trigger your allergy symptoms.

Allergy shots typically involve two phases, buildup and maintenance.

It is an escalating dose immunotherapy, when you gradually increase the exposure level of the identified allergen.

The buildup phase lasts for 3 to 6 months. You receive shots 1 to 3 times a week. The doctor will gradually increase the amount of allergen in each shot to help your body build tolerance.

In the maintenance phase you need shots less frequently, usually about once a month. This phase can continue for 3 to 5 years or even longer depending on your progress.

I was never interested in allergy shots because there are so many injections needed.

I found EPD of interest because you take just two shots a year and the effect may potentially control the allergy after 2 or 3 years.

EPD is not expensive and I suppose that is why nobody wanted to invested the tens of millions of dollars to get approval by the FDA. It remains approved for use in the UK, which is ultra conservative when it comes to medicines.

Carmello Rizzo is offering EPD in Italy and elsewhere.


Gene therapy for autism?

I did go to a presentation with an interesting title:

Developing effective therapeutics for Autism Spectrum Disorder

It was not really what I was expecting. It was a young MIT researcher talking about the potential to develop gene therapies to replace mutated genes with a new ones. They are doing this in a model of autism caused by a mutated copy of the SHANK3 gene.

I called him Dr Viral Vector and did have a chat with him. The most interesting thing about his technology is that not only can he target a specific type of cell, but he can target a specific part of the brain, or indeed any part of the body.

At the moment they inject a virus carrying the new gene directly into the brain. That is not going to go down so well with human subjects. The next stage is to try injecting the virus into a vein.

I did talk about the two gene therapies for Rett syndrome now in human trials in my presentation. The ultimate problem is the likely $3 million cost. 

You can use gene therapy as an immunotherapy. 



At the conference I was asked about a gene called DCLRE1C, it encodes the DCLRE1C protein, also known as Artemis.


Artémis (Diane), the huntress. Roman copy of a Greek statue, 2nd century. Galleria dei Candelabri

Source: By Jean-Pol GRANDMONT - Own work, CC BY-SA 3.0,


The Artemis protein is named after the Greek goddess Artemis, who was associated with the hunt, wilderness, wild animals, childbirth, and protection. This connection likely comes from the crucial role Artemis plays in DNA repair, which is essential for maintaining the integrity of the genetic material, like a protector safeguarding the building blocks of life.

Complete loss of function in DCLRE1C typically causes severe combined immunodeficiency. This is called Artemis-deficient severe combined immunodeficiency (ART-SCID).

Fortunately many possible mutations only partially impair the function of the DCLRE1C gene. They can lead to a spectrum of conditions, including atypical SCID, Omenn syndrome, Hyper IgM syndrome, and even just antibody deficiency. These conditions may have milder symptoms compared to classic SCID.

IVIG is a beneficial therapy for immunodeficiency; but is very expensive and not curative.

Humans all have 2 copies of the DCLRE1C and it is theoretically possible to increase expression of the good copy. But that is another story.


A gene therapy already exists for full-on ART-SCID.

Lentiviral Gene Therapy for Artemis-Deficient SCID

Why not use it in less severe cases?

The problem is going to be money, both for a lifetime on IVIG or a “hopefully” one-off gene therapy.

One lady in the audience of my talk had herself taken an expensive gene therapy and was not impressed.


Other interesting presentations

Pierre Drapeau from McGill University spoke about trying to repurpose a cheap old drug, called Pimozide, to treat motor neuron disease /ALS.  This was interesting because the process is similar to repurposing a drug for autism.

Pimozide is an old antipsychotic drug and it seems to work in ALS through its effect on a type of calcium channel called the T-type. Yes, just as in much autism, calcium channels are misbehaving.

The drawback of Pimozide is that it also blocks dopamine receptors in the brain, which is good if you have Tourette’s, but if you have ALS you then get symptoms of Parkinson’s as a side effect.

The solution is to tinker with the molecule and find a version (an analog) that will do the business with the T channels without causing tremors.  It looks like, via trial and error, this is nearly solved.

The whole process has already been going on for many years, it will take many more.

Life expectancy with ALS is only 2-5 years and they struggle to find test subjects in Canada. It looks like they may do trials in China.


An eye opener

A presentation with a very hard to digest title was also an eye opener. You can take a picture of the cornea in your eye and accurately diagnose all kinds of disorders. They started with peripheral neuropathy in diabetics and most recently moved on to people with autism. Using artificial intelligence (AI) they can now make a diagnosis just based on the nerve loss they observe in the cornea. They also can potentially measure the effect of therapies by the regeneration of those nerve fibers.  This is really clever. When Rayaz Malik started down this path, all the neurologists thought he was mad. Many years later and corneal confocal microscopy is widely used around the world, but not yet for autism diagnosis.

Antonio Persico is a well known autism clinician, he appeared virtually. He was mainly talking about antipsychotics. I had expected rather more. 



Immunotherapy addresses one of the four problem areas in autism. There cannot be a one size fits all approach, but you can certainly try camel milk. Addressing food allergy and intolerance is relatively straightforward and you do not need any fancy expensive genetic testing, as Carmello Rizzo pointed out.

There are people for whom genetic testing and/or a spinal tap opens the door to a precise diagnosis and hopefully treatment. That proved to be an unexpected controversial issue in my presentation.

My talk at the conference was all about using personalized medicine to treat autism. The organizer of the event reads this blog and knows that I am rather an outsider, since I am more in treating autism than just researching it.

I had a two and a half hour time slot and I made sure to use it all. 

Advances in Personalized Medicine to Treat Autism

I should mention that I also had some long conversations with Paul Shattock, who pretty much founded the gluten and casein free diet years ago, back at the University of Sunderland. If you are interested in the history of autism, he is a great person to talk to. He is nearly 80 years old, but still has a sharp sense of humour. He has stumbled into more than his fair share of controversies. In Abu Dhabi his opinions and observations were widely shared by other speakers. One younger American speaker thought his views were dangerous; had he taken the time to talk to Paul, he would have found them pretty well thought out. I did ask Paul what has happened to his old friend Andew Wakefield – apparently making another film.



Thursday 24 June 2021

Betaine (TMG) and Gene Therapy as potential alternatives to Bumetanide Treatment in Autism?

Betaine (also known as TMG, or trimethylglycine) is a methyl derivative of glycine, first isolated from sugar beet and hence its name.

Today’s post was prompted by our reader, and Covid home-school instructor, AJ.  He raised the question of whether betaine can be used like Bumetanide to normalize chloride levels in neurons.

I am combing this idea with news from Genoa in Italy, where they have developed gene therapy as an alternative to Bumetanide and in their words :-

“This sets the stage for the development of a gene therapy approach to overcome the shortcomings of bumetanide treatment.”

The interesting thing is that neither of these ideas come from autism research.  The idea to use Betaine was stumbled upon and was then written up in a Norwegian case study about Creatine transporter deficiency.  The Italians are trying to improve cognition in brain disorders and their model of choice was Down syndrome. 

As we have seen time and again, elevated chloride within neurons is a common feature of many types of brain disorders from some idiopathic autism, to Down syndrome, to adult conditions such as Parkinson’s disease.  Today we learn that it is may well be a feature of Creatine Transporter Deficiency.

I have been rather wary of writing about any kind of gene therapy, because it seemed either too far ahead of its time, or just absurdly expensive.  There are some new $1+ million treatments.

This may be about to change given that the Biontech (AKA Pfizer vaccine), Moderna, Janssen (Johnson & Johnson) and Oxford AstraZeneca vaccines for Covid 19 are all based on gene therapy.

The Biontech people are really clever and were already trying to treat various kinds of cancer and other condition using gene therapy, before they developed their highly successful Covid vaccine.

The Italians in Genoa used an adeno-associated virus (AAV)-mediated RNA interference (RNAi) to target and reduce neuronal NKCC1 expression, rescue neuronal Cl-  homeostasis, GABAergic transmission, and cognitive deficits.   The benefit was still there 6 months after the injection.

Don’t worry if the above paragraph makes little sense. Just read on.

The same type of adeno-associated virus (AAV) vector is the platform for gene therapy delivery used in the Astra Zeneca, Janssen and the Russian Sputnik covid vaccines.

The virus is just the delivery system (vector) to get some genetic code into cells.

The Oxford-AstraZeneca COVID-19 vaccine uses a chimpanzee adenoviral vector. It delivers the gene that encodes the SARS-CoV-2 spike protein, to our cells.  Our cells then transcribe this gene into messenger RNA, or mRNA, which in turn prompts our cellular machine to make the spike protein in the main body of the cell. The mRNA molecule behaves essentially like a recipe.  Then our cells present the spike protein on the cell surface, prompting our immune system to make antibodies and mount T cell responses.

Biontech and Moderna are pioneers of mRNA vaccines, which bypass one step in the above process. They do not require our cells to make the messenger RNA, or mRNA.  They have already made it for you.


Gene therapy for autism?

Single gene autisms are all potential candidates for gene therapy.

The problem is that most autism and all Down syndrome is polygenic, there can be hundreds of miss-expressed genes.

But the researchers in Italy show us that even polygenic autism and Down syndrome can benefit from therapy targeting a single gene.  You just have to select the right one.

The problem is the price. Covid vaccines are made in huge quantities and are cheap.

Customized gene therapy is ultra expensive, in part because each therapy has to be approved individually.


An NKCC1 Gene Therapy?

The Italians have already made the NKCC1 Gene Therapy.  The question is will it ever going be available to humans with Down Syndrome, Autism or even Parkinson’s disease?

Restoring neuronal chloride homeostasis with anti-NKCC1 gene therapy rescues cognitive deficits in a mouse model of Down syndrome

A common feature of diverse brain disorders, is the alteration of GABA-mediated inhibition due to aberrant intracellular chloride homeostasis induced by changes in the expression and/or function of chloride transporters. Notably, pharmacological inhibition of the chloride importer NKCC1 is able to rescue brain-related core deficits in animal models of these pathologies and some human clinical studies. Here, we show that reducing NKCC1 expression by RNA interference in the Ts65Dn mouse model of Down syndrome (DS) restores intracellular chloride concentration, efficacy of GABA-mediated inhibition and neuronal network dynamics in vitro and ex vivo. Importantly, AAV-mediated neuron-specific NKCC1 knockdown in vivo rescues cognitive deficits in diverse behavioral tasks in Ts65Dn animals. Our results highlight a mechanistic link between NKCC1 expression and behavioral abnormalities in DS mice, and establish a molecular target for new therapeutic approaches, including gene therapy, to treat brain disorders characterized by neuronal chloride imbalance.


This sets the stage for the development of a gene therapy approach to overcome the shortcomings of bumetanide treatment.

This highlights a causative role of NKCC1 upregulation in learning and memory deficits in adult Ts65Dn mice, thus also validating brain NKCC1 as a target for ameliorating cognitive disabilities in DS. Furthermore, our neuro-specific knockdown approach points to neurons as major players in the NKCC1- dependent cognitive impairment in DS mice. Nevertheless, we cannot exclude that other cell types which also express NKCC1 (e.g. glial cells) could still play a role in the overall cognitive impairment that characterizes DS.

Despite the very large and fast-increasing literature both on animal models and patients indicating positive outcomes upon bumetanide treatment, there is not yet a strong demonstrated direct link between NKCC1 inhibition, restoration of Cl- homeostasis and full GABAergic inhibitory signaling, and rescue of brain deficits.  Moreover, bumetanide has strong diuretic activity, triggering ionic imbalance, and potential ototoxicity 25,26.  This hampers its use for clinical applications in lifelong treatments4,27 and may strongly jeopardize treatment compliance along years of treatment.  Moreover, bumetanide was given systemically in most studies, and the suboptimal brain pharmacokinetic profile of the drug28 raises questions on its mechanism of action29.  Here, we demonstrate that adeno-associated virus (AAV)-mediated RNA interference (RNAi) to target (and reduce) neuronal NKCC1 expression rescues neuronal Cl- homeostasis, GABAergic transmission, and cognitive deficits in the Ts65Dn mouse model of Down syndrome. This sets the stage for the development of a gene therapy approach to overcome the shortcomings of bumetanide treatment.


“Thus, our results indicate the efficacy of long-term AAV9-mediated neuro-specific NKCC1 knockdown in rescuing cognitive deficits in Ts65Dn mice.”


“Besides establishing a causal link between NKCC1 upregulation and cognitive impairment in DS, our data also provide a proof-of-concept for a neuro-specific RNAi gene therapy approach to restore hippocampus-dependent cognitive behaviors in adult animals specifically in the brain, and without affecting peripheral organs (e.g., the kidney). This is particularly relevant in the context of the current clinical trials repurposing the strong diuretic bumetanide to treat brain disorders with impaired chloride homeostasis3.  Importantly, we achieved a comparable degree of long-term cognitive rescue with two different amiR sequences against NKCC1, underlining the specificity of our approach.”


Gone Fishing

If a trip to Italy for gene therapy is not realistic, this takes us back to AJ’s idea, which is to use Betaine.  The correct version is TMG or glycine betaine, and confusingly not Betaine HCl.

Fish love the taste of betaine.

Betaine was first isolated from sugar beet.

I recall from my time at the sugar factory, when I was 18, that once you have sliced up the sugar beet and extracted as much sugar as possible you are left with the pulp.  This pulp is dried, molasses is added back and then it is made into pellets.  The pellets are fed to cattle and horses.  They taste pretty bad in my opinion.

To humans it tastes bad because of the beet molasses by-product.

The molasses by-product from sugar cane tastes great to humans.  That is why they make rum in the Caribbean, and not in England or Canada.

Brown sugar from a sugar beet factory is made by adding sugar cane molasses to white sugar from beet.  It is a cheat really.

Cows love sugar beet by-products.

It turns out that fish love betaine HCl.

Betaine HCl is an excellent natural attractor that stimulates a strong, prolonged feeding response from carp and many other coarse fish.

Betaine HCl is now used to induce feeding in the fish farming industry

As our reader Tyler has highlighted, Betaine HCl, that fish like and is available is a cheap supplement is not the same as the Betaine used in the medical case study. Confusingly, the original Betaine (TMG, or called glycine betaine) gave way to a class of compounds all called betaines. One of these betaines is betaine HCL.

In most cases, in the medical literature when they refer to Betaine, they mean glycine betaine, also known as TMG.

Betaine HCl is used to increase acidity in your stomach. The effect of betaine compounds other than glycine betaine/TMG on NKCC1 is unknown.

Glycine Betaine (TMG) and NKCC1

It seems that betaine reduces your level of NKCC1 RNA. 

In your DNA are the instructions to make the NKCC1 transporter. To go from these instructions to actually making the transporters you need RNA.

In some autism there are too many NKCC1 transporters, so put simply there was too much NKCC1 RNA. So, if you can find a substance that reduces NKCC1 RNA, you might well solve the problem.

The caveat is that the substance must not also increase KCC2 RNA.  This appears to be what taurine does.

Here, finally, is AJ’s paper:

Treatment experience in two adults with creatine transporter deficiency


Creatine transporter deficiency (CTD) is an X-linked form of intellectual disability (ID) caused by SCL6A8 mutations. Limited information exists on the adult course of CTD, and there are no treatment studies in adults.


We report two half-brothers with CTD, 36 and 31 years at intervention start. Their clinical phenotypes were consistent with CTD, and intervention was indicated because of progressive disease course, with increased difficulties speaking, walking and eating, resulting in fatigue, and malnutrition. We therefore performed treatment trials with arginine, glycine and a proprietary product containing creatine and betaine, and then a trial supplementing with betaine alone. Results In the older patient, glycine and arginine were accompanied by adverse effects, while betaine containing proprietary product gave improved balance, speech and feeding. When supplementation stopped, his condition deteriorated, and improved again after starting betaine supplement. Betaine supplementation was also beneficial in the younger patient, reducing his exhaustion, feeding difficulties and weight loss, making him able to resume his protected work.

Discussion & conclusion

We report for the first time that betaine supplement was well tolerated and efficient in adults with CTD, while arginine and/or glycine were accompanied by side effects. Thus, betaine is potentially a new useful treatment for CTD patients. We discuss possible underlying treatment mechanisms. Betaine has been reported to have antagonistic effect on NKCC1 channels, a mechanism shared with bumetanide, a medication with promising results in both in autism and epilepsy. Further studies of betaine's effects in well-designed studies are warranted.


The mechanism of betaine’s assumed favorable effect is unknown. We do not know whether betaine influences the cell creatine content in itself or its effects are more aspesific. However, we would like to present some hypotheses. First, betaine may have effect in CTD by modulating GABA-transmission. Betaine has been reported to have an antagonistic effect on NKCC1 channels, which also influences GABAergic neurotransmission. Inhibiting NKCC1 is a mechanism shared with bumetanide, a well-known diuretic medication that in recent years has been found to influence GABAergic transmission, and thereby it has been found promising in treatment of several brain conditions, including autism, and epilepsy. NKCC1 inhibition by bumetanide has also been tried with success in other rare neurodevelopmental disorders fragile X syndrome and tuberous sclerosis. Second, betaine’s properties as an osmolyte may be of importance, as betaine has similarities with creatine in being an osmolyte. Osmotic properties are thought to be one of the central mechanism behind bumetanide’s efficacy in treating brain disorders. Thus, it could be speculated that the lack of intracellular creatine in CTD may result in inefficient osmolyte regulation, and that betaine supplementation replaces the lacking creatine and thereby improves the neuronal adaption to salinity changes, edema or cellular dehydration. Betaine has osmolyte properties that even makes it act as a “chemical chaperone” increasing the stability of cell and membrane proteins. Fourth, it is possible that betaine has some effect through modifying methylation. Methylation of GAA by GAMT to form creatine is a rate-limiting step in the creatine synthesis by neurons. Betaine could stimulate this by donating methyl groups to SAMe, which donates a methyl group to GAA to form creatine. This might reduce the burden when body demands more methyl groups for creatine synthesis. Similar mechanisms may be responsible for a beneficial effect of both betaine and s-adenosyl methionine (SAMe). However, as creatine and GAA share the same transporter, one would not expect GAA to enter the GAMTexpressing cells in patients suffering from CTD. Still, it cannot be excluded that there is some rest function in the creatine transporter, and that increased endogenous synthesis improves the condition slightly. Furthermore, it is possible that CTD increases the need for methylation agents in general, as creatine supplementation has been found to reduce the need for other methylation agents [34]. Thus, it is likely that betaine may have a positive effect in CTD by improving methylation capacity for other reactions than those directly involved in creatine production. Betaine’s effect on muscle may be also of importance, as animal studies have shown that muscles growth improves with betaine [35], which potentially could have had a positive impact on our patients fatigue and weight loss. To summarize, betaine has several properties that make it likely that it will have a beneficial effect in CTD, especially the properties as an osmolyte, a down regulator of the NKCC1 channel and an influencer of GABAergic transmission. These properties are similar to the properties of bumetanide, a promising new medication for treatment of autism and epilepsy, which are common symptoms of CTD. Further research is needed, however, to elucidate the role of betaine in CTD.

If you read the detail of the old paper that is referred to in the above paper, you see that betaine is not blocking the NKCC1 channels as suggested, but it seems to be reducing the number of them.  The net effect may be the same, but the process is very different.


Expression and regulation of the Na+/K+/2Cl− cotransporter NKCC1 in rat liver and human HuH-7 hepatoma cells

The expression of sodium potassium chloride cotransporter 1 (NKCC1) was studied in different liver cell types. NKCC1 was found in rat liver parenchymal and sinusoidal endothelial cells and in human HuH-7 hepatoma cells. NKCC1 expression in rat hepatic stellate cells increased during culture-induced transformation in the myofibroblast-like phenotype. NKCC1 inhibition by bumetanide increased α1-smooth muscle actin expression in 2-day-cultured hepatic stellate cells but was without effect on basal and platelet-derived-growth-factor-induced proliferation of the 14-day-old cells. In perfused rat liver the NKCC1 made a major contribution to volume-regulatory K+ uptake induced by hyperosmolarity. Long-term hyperosmotic treatment of HuH-7 cells by elevation of extracellular NaCl or raffinose concentration but not hyperosmotic urea or mannitol profoundly induced NKCC1 mRNA and protein expression. This was antagonized by the compatible organic osmolytes betaine or taurine. The data suggest a role of NKCC1 in stellate cell transformation, hepatic volume regulation, and long-term adaption to dehydrating conditions.


Aha!  Glycine Betaine and Taurine – not so fast 

You have to check the effect on both NKCC1 and KCC2.  One lets chloride into neurons and the lets it out.  You want to block NKCC1 and not KCC2, otherwise you undo all the good you have done.

Both glycine betaine (TMG) and taurine are already used as autism supplements at low doses.  The paper below suggest that Taurine is not a good idea for people with high levels of chloride within neurons.


Taurine inhibits K+-Cl- cotransporter KCC2 to regulate embryonic Cl- homeostasis via with-no-lysine (WNK) protein kinase signaling pathway

GABA inhibits mature neurons and conversely excites immature neurons due to lower K(+)-Cl(-) cotransporter 2 (KCC2) expression. We observed that ectopically expressed KCC2 in embryonic cerebral cortices was not active; however, KCC2 functioned in newborns. In vitro studies revealed that taurine increased KCC2 inactivation in a phosphorylation-dependent manner. When Thr-906 and Thr-1007 residues in KCC2 were substituted with Ala (KCC2T906A/T1007A), KCC2 activity was facilitated, and the inhibitory effect of taurine was not observed. Exogenous taurine activated the with-no-lysine protein kinase 1 (WNK1) and downstream STE20/SPS1-related proline/alanine-rich kinase (SPAK)/oxidative stress response 1 (OSR1), and overexpression of active WNK1 resulted in KCC2 inhibition in the absence of taurine. Phosphorylation of SPAK was consistently higher in embryonic brains compared with that of neonatal brains and down-regulated by a taurine transporter inhibitor in vivo. Furthermore, cerebral radial migration was perturbed by a taurine-insensitive form of KCC2, KCC2T906A/T1007A, which may be regulated by WNK-SPAK/OSR1 signaling. Thus, taurine and WNK-SPAK/OSR1 signaling may contribute to embryonic neuronal Cl(-) homeostasis, which is required for normal brain development.


So, it is likely only Glycine Betaine (TMG) may be of potential benefit, in the case of lowering chloride.


Glycine Betaine in the broader research


Betaine in Inflammation: Mechanistic Aspects and Applications

Betaine is known as trimethylglycine and is widely distributed in animals, plants, and microorganisms. Betaine is known to function physiologically as an important osmoprotectant and methyl group donor. Accumulating evidence has shown that betaine has anti-inflammatory functions in numerous diseases. Mechanistically, betaine ameliorates sulfur amino acid metabolism against oxidative stress, inhibits nuclear factor-κB activity and NLRP3 inflammasome activation, regulates energy metabolism, and mitigates endoplasmic reticulum stress and apoptosis. Consequently, betaine has beneficial actions in several human diseases, such as obesity, diabetes, cancer, and Alzheimer’s disease.


Betaine is a stable and nontoxic natural substance. Because it looks like a glycine with three extra methyl groups, betaine is also called trimethylglycine . In addition, betaine has a zwitterionic quaternary ammonium form [(CH3)3N+ CH2COO−] (Figure 1). In the nineteenth century, betaine was first identified in the plant Beta vulgaris. It was then found at high concentrations in several other organisms, including wheat bran, wheat germ, spinach, beets, microorganisms, and aquatic invertebrates. Dietary betaine intake plays a decisive role in the betaine content of the body. Betaine is safe at a daily intake of 9–15 g for human and distributes primarily to the kidneys, liver, and brain. The accurate amount of betaine intake generally relies on its various sources and cooking methods. Besides dietary intake, betaine can be synthesized from choline in the body. Studies report that high concentrations of betaine in human and animal neonates indicate the effectiveness of this synthetic mechanism.


Boosting amino acid derivative may be a treatment for schizophrenia

Many psychiatric drugs act on the receptors or transporters of certain neurotransmitters in the brain. However, there is a great need for alternatives, and research is looking at other targets along the brain's metabolic pathways. Lack of glycine betaine contributes to brain pathology in schizophrenia, and new research shows that betaine supplementation can counteract psychiatric symptoms in mice.



Supplement treats schizophrenia in mice, restores healthy “dance” and structure of neurons Repurposed drug works by building cells’ skeleton and transportation network




Early on in the Covid saga, I saw interviews with both the Moderna researchers and the Oxford (AstraZeneca) researchers. Both claimed that they designed their vaccines over a weekend.  This was made possible by the Chinese releasing the DNA code of the virus.

When you think about gene therapy for autism and Down syndrome, the same likely applies; much could be achieved over a weekend.

The expensive and time-consuming part is the testing and approval process.

In the Covid pandemic the approval process was modified to allow for emergency use.  Perhaps this should also be the case for all gene therapies?

What use is a $2 million therapy for autism or Down syndrome?

In theory, if you gave your gene therapy prior to birth or shortly thereafter, it might be fully curative.  Realistically, by the time you get the therapy it is just going to be beneficial and you will still need other ongoing therapies.

Note that gene therapy normally applies to just one gene.  In Down syndrome people have a third copy of all, or just part, of Chromosome 21.  This results directly in the miss-expression of hundreds of genes from that chromosome.

The gene that encodes NKCC1 is on Chromosome 5, which has nothing directly to do with Down syndrome.

The NKCC1 transporter is over-expressed in Down syndrome as a down stream consequence of the disorder. It is caused by the “faulty GABA switch”, referred to in earlier posts.

The Italian gene therapy to lower chloride in neurons and so raise cognition, has numerous applications, in people currently of all ages, so there is a big potential market.

Why not gene therapy for all single gene autisms?  It could be a highly productive use of the researcher’s weekends, for a year or two.

The issue is who would pay for the $20 to $30 million approval process, for each gene?

Maybe some of the billions in profit from clever Covid vaccines could be used for pro bono gene therapy?  Highly unlikely.

Biontech, who are the brains behind the Pfizer vaccine, do have plans to develop gene therapy for other medical conditions.  I think these will be ultra expensive,

That brings me back to Glycine Betaine (TMG), is 10g a day of this supplement really going to reduce the expression of NKCC1 transporters in neurons and so lower chloride within neurons?  It seems to work in creatine transporter deficiency, is all we can say.  

Glycine betaine, at much lower doses, has been used by DAN and now MAPS doctors for decades. They use it as a “methyl-donor”.  There is a combination of real science and hocus-pocus surrounding DNA methylation. 

 DNA Methylation and Susceptibility to Autism Spectrum Disorder