Altered TRPM7-Dependent Calcium Influx in Natural Killer Cells of Myalgic Encephalomyelitis/Chronic Fatigue Syndrome Patients

[SWAlexander]

Please note NCNED Griffith University will be holding an international ME Conference 9-10 November, 2023 at Kingscliffe, New South Wales, Australia. Patients are welcome to attend and can ask questions.

https://twitter.com/i/web/status/1691705098014245255

Abstract​

Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a disabling multisystemic condition. The pathomechanism of ME/CFS remains unestablished; however, impaired natural killer (NK) cell cytotoxicity is a consistent feature of this condition. Calcium (Ca2+) is crucial for NK cell effector functions. Growing research recognises Ca2+ signalling dysregulation in ME/CFS patients and implicates transient receptor potential ion channel dysfunction. TRPM7 (melastatin) was recently considered in the pathoaetiology of ME/CFS as it participates in several Ca2+-dependent processes that are central to NK cell cytotoxicity which may be compromised in ME/CFS. TRPM7-dependent Ca2+ influx was assessed in NK cells isolated from n = 9 ME/CFS patients and n = 9 age- and sex-matched healthy controls (HCs) using live cell fluorescent imaging techniques. Slope (p < 0.05) was significantly reduced in ME/CFS patients compared with HCs following TRPM7 activation. Half-time of maximal response (p < 0.05) and amplitude (p < 0.001) were significantly reduced in the HCs compared with the ME/CFS patients following TRPM7 desensitisation. Findings from this investigation suggest that TRPM7-dependent Ca2+ influx is reduced with agonism and increased with antagonism in ME/CFS patients relative to the age- and sex-matched HCs. The outcomes reported here potentially reflect TRPM3 dysfunction identified in this condition suggesting that ME/CFS is a TRP ion channelopathy.

https://www.mdpi.com/2218-273X/13/7/1039

 

[datadragon]

Interesting. Dysregulation of TRP melastatin subfamily members (TRPM) and calcium signalling processes are implicated as being involved. In this earlier review here for example, they presented TRPM7 as a potential candidate in the pathomechanism of ME/CFS (one part), as TRPM7 is increasingly recognized as a key mediator of physiological and pathophysiological mechanisms affecting neurological, immunological, cardiovascular, and metabolic processes inherent in ME/CFS. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8535478/

What I may add is that I found Zinc regulates TRPM7's binding to transcription factors and TRPM7's kinase activity. Further, it is also being shown to be required for the inhibition of TRPM7 by intracellular magnesium so it appears any loss of zinc could be a culprit. On that note, Zinc absorption becomes reduced due to a lowering of zinc uptake in the gut, and also becomes further unavailable to utilize during inflammation and infection as well due to metallothionein, which sequesters zinc after inflammatory cytokines. The activity of the TRPM7 channel is negatively regulated by intracellular Mg2+. We previously reported that oxidative stress enhances the inhibition of TRPM7 by intracellular Mg2+. The zinc-binding motif is essential for the intracellular Mg2+-dependent regulation of the TRPM7 channel activity by its kinase domain and that the cysteines in the zinc-binding motif play a role in the oxidative stress response of TRPM7.
https://pubmed.ncbi.nlm.nih.gov/33999118/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5544332/
https://pubmed.ncbi.nlm.nih.gov/28696294/

Normally TRPM7 is induced by IL-18. IL-18 increased TRPM7 expression through ERK1/2 (extracellular signal-regulated kinase 1/2) signaling activation, and TRPM7 currents were augmented by IL-18 treatment. https://pubmed.ncbi.nlm.nih.gov/28860220/ The activation of the NLRP3 inflammasome (inflammation), results in the processing and secretion of active Interleukin 1β (IL-1β) and IL-18. Type I IFN signaling is necessary for the induction of IL-18, but not IL-1β, which points to a critical and differential role for type I IFN in regulating IL-18 signaling. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5544497/ Interleukin-18 (IL-18) synergizes with IL-2 to enhance cytotoxicity, interferon-gamma (IFN-y) production, and expansion of natural killer cells https://pubmed.ncbi.nlm.nih.gov/11221875/

Complement subsequently elicits secretion of both IL-1β and IL-18 in vitro and in vivo via further activation of the NLRP3 inflammasome. https://pubmed.ncbi.nlm.nih.gov/23817414/

Extracellular ATP can cause P2X receptors to activate the NOD-like receptor 3 (NLRP3) inflammasome and cause IL-1β and IL-18 maturation and release. https://pubmed.ncbi.nlm.nih.gov/23434541/

Homocysteine activated NLRP3 inflammasomes in THP-1-differentiated macrophages and promoted subsequent production of IL-1β and IL-18 in macrophages, which were blocked by NLRP3 gene silencing or the caspase-1 inhibitor Z-WEHD-FMK. https://pubmed.ncbi.nlm.nih.gov/28394319/

TRPM7 protein levels were also significantly increased in the HypoMg-medium treated macrophages https://www.cell.com/biophysj/fulltext/S0006-3495(22)01694-0

 

[SWAlexander]

Last year in the midst of acute C-19 symptoms I have taken mineral and primary Zinc as a primary supplement. I experienced some improvement (reduced inflammation) but also side effects such as:
Gastrointestinal problems, temp. loss of taste and allergic reactions.
Later I learned it is possible that copper deficiency was due to Zinc interference. A copper deficiency can lead to anemia, fatigue, and neurological problems.

Then, I switched to copper for 10 days. Copper is a component of several proteins that are involved in the immune system, such as the antioxidant enzyme superoxide dismutase, preventing collagen (my major problem with APS) and myelin reduction.

Both zinc and copper are vital minerals. But I recommend first a blood test before taking a mineral supplement to avoid eventual imbalance.

 

[datadragon]

Later I learned it is possible that copper deficiency was due to Zinc interference. A copper deficiency can lead to anemia, fatigue, and neurological problems.

Then, I switched to copper for 10 days. Copper is a component of several proteins that are involved in the immune system, such as the antioxidant enzyme superoxide dismutase, preventing collagen (my major problem with APS) and myelin reduction.

Both zinc and copper are vital minerals. But I recommend first a blood test before taking a mineral supplement to avoid eventual imbalance.

Copper and Zinc are metabolic antagonists - Zinc maintains a balance with Copper in the blood, where changes in these two trace elements tend to be inversely related. A low plasma Zinc concentration is nearly always associated with a high serum Copper concentration which happens after chronic inflammation. Wapnir and Balkman (1991) investigated these concepts using a duodenal-jejunal single-pass perfusion process in rats. They found that Cu absorption decreased with increasing Zn presence. So in general its more about balance.

However, ceruloplasmin production needed to make copper and iron usable (bio available) has been shown to be regulated by 13-cis retinoic acid, a metabolite of Vitamin A https://www.ncbi.nlm.nih.gov/pubmed/3655940 which requires zinc in Vitamin A metabolism (and Vitamin A is involved in zinc metabolism). So again more about balance and much more common to have lower zinc, higher copper that becomes 'free' and therefore causes a deficiency at the same time despite having plenty of copper intake. Its like being in the middle of the ocean surrounded by water yet still not have water to drink. So you would have to check if you are experiencing a 'deficiency' due to low ceruloplasmin and actually have higher unbound copper and likely dealing with some form of chronic inflammation, or if you actually had a true deficiency of copper which is far less common. Its possible to check serum copper and serum ceruloplasmin (the usable copper) and can calculate free copper levels unbound to ceruloplasmin but this is a spot check at the time of the test only and does not reflect any long term accumulation in the tissues that may have also been going on. https://web.archive.org/web/2021030...ients-families/lab-tracker-copper-calculator/

 

[SWAlexander]

Vitamin A

This could be the key. Thanks.
Retinoic acid is a form of vitamin A. It may help the ever present inflammation, improve collagen and my scaling psoriasis.

 

[datadragon]

Vitamin A

This could be the key. Thanks.
Retinoic acid is a form of vitamin A. It may help the ever present inflammation, improve collagen and my scaling psoriasis.

Vitamin A palmitate is a form of vitamin A. It’s found in animal products, such as liver, eggs, and cheese. It’s also called preformed vitamin A and retinyl palmitate. Vitamin A retinyl palmitate is available as a manufactured supplement and vitamin A palmitate is a retinoid (retinol). There’s a common misunderstanding that beta-carotene found in fruits and vegetables is the same thing as vitamin A. Beta Carotene can be converted to retinol by the enzyme β-carotene 15,15′-monooxygenase (BCMO1 gene) and is then used by the body in the same way as preformed vitamin A from animal products is used or stored, but almost half of everyone have variants (two common mutations) on the BCMO1 gene which cause a 30% to 70% decrease in the amount of vitamin A that we can convert from beta-carotene. This means that contrary to popular wisdom, vegetables like carrots and red peppers may not be adequate food sources of vitamin A for many. This may be quite normal in some people however as your body has individual genetics that help to balance inflammation/immunity and the levels and so have genes in different areas modified that help with that. Retinol however then needs to be converted by enzymes into other active metabolites such as retinoic acid, and I found that both zinc and cytochrome P450 enzymes that require magnesium as a cofactor are involved in these additional conversions and also prevent excessive levels, Perhaps a low ceruloplasmin level is among ways to note if there are problems converting to the active forms.

Zinc and Vitamin A are both lowered from inflammation and infection, especially with fever. Episodes of acute infection deplete body stores of vitamin A. We have found that significant amounts of retinol and retinol-binding protein (RBP) were excreted in the urine during serious infections, whereas only trace amounts were found in the urine of healthy control subjects. Subjects with fever (temperature > or = 38.3 degrees C) excreted significantly more retinol (geometric mean = 1.67 mumol/d) than did those without fever. https://pubmed.ncbi.nlm.nih.gov/8074070/ Severe infections in adult patients (i.e., sepsis and pneumonia) result in excretion of large quantities of Vitamin A retinol in the urine and depletion of vitamin A stores. https://www.ncbi.nlm.nih.gov/pubmed/1152102/ https://www.ncbi.nlm.nih.gov/pubmed/7762530/ Retinol levels decline rapidly as part of the acute phase response https://www.ncbi.nlm.nih.gov/pubmed/11063445/ and this translates into increased susceptibility to infection, creating a “vicious circle” difficult to break https://www.ncbi.nlm.nih.gov/pubmed/10466190/ Vitamin A insufficiency is associated with increased mortality to lung infections and immune responses to infection were compromised upon loss of Vitamin A. https://www.medicalnewstoday.com/articles/219513#1

Vitamin A is involved in zinc metabolism and vice versa. For example, Zinc deficiency is thought to interfere with vitamin A metabolism in several ways: (1) Zinc deficiency results in decreased synthesis of retinol-binding protein (RBP), which transports retinol through the circulation to peripheral tissues for its utilization by the body and protects the organism against potential toxicity of retinol; (2) zinc deficiency results in decreased activity of the enzyme that releases retinol from its storage form, retinyl palmitate, in the liver; and (3) zinc is required for the enzyme that converts retinol into retinal. Zinc is involved in that conversion of retinol to retinaldehyde (retinal) and retinal to retinoic acid, respectively - The zinc metalloenzyme ADH is required for this oxidative process. Zinc uptake is lowered and also becomes less available to utilize during chronic inflammation/infection.

The gene encoding alcohol dehydrogenase-1 (Adh1) (which requires Zinc.) greatly facilitates degradative metabolism of excess Vitamin A retinol into retinoic acid which may help protect against toxic effects of high dietary vitamin A intake or supplementation. and also helps convert to the active forms. https://portlandpress.com/biochemj/...lular-retinol-binding?redirectedFrom=fulltext

Since there are no known enzymes that can reduce retinoic acid to retinal, excessive or unneeded retinoic acid is not recycled back to retinol/retinyl ester and must be catabolized and eliminated from the body. This catabolism is catalyzed by one of several cytochrome P450 (CYP) enzymes which require magnesium. Any factor that lowers magnesium or cytochrome P450 enzymes can lead to excessive retinoic acid. https://www.jlr.org/article/S0022-2275(20)42124-8/fulltext

Over 600 enzyme systems require Magnesium as a cofactor to function optimally, including the cytochrome P450 enzymes (CYP450 https://www.liebertpub.com/doi/abs/10.1089/pai.1994.8.7

I just mention that as it may require lowering of inflammation beyond just taking supplements sometimes to restore function due to the impact inflammation and infection has on zinc and cytochome p450 enzymes.

 

[heapsreal]

Something I've often thought about with low nk function in cfsme, is it the cause of cfsme ie people had low nk function prior to the onset of cfs and then caught an infection they didn't seem to recover from. Not saying they still have the infection, they could but also the triggering infection may have been able to cause more damage when one has low nk function but still be in the hit and run theory.
Or is the low nk function like an immune depletion type of scenario. If someone has chronic reactivating virus or other infections, this could be putting a strain on the nutrients needed eg bone marrow production, natural interferon needed for nk cells etc and this lowers nk function and or numbers?? Sort of how HIV depletes pts cd4 T cells but in cfsme it's depleting pts nk function.
I guess a third possibility is the initial trigger has damaged the immune system in some way. I guess it's hard to answer as I don't think there are any studies of a group of people without cfsme who have had their nk function tested and later ended up with cfs and had nk tested, so had before and after cfsme nk function compared. Probably be a hard study to set up and require alot of people.

 

[Violeta]

"Furthermore, the conversion of β-carotene to retinol is inhibited in patients with hypothyroidism, and a negative relationship persists between β-carotene and retinol."

I do find that vegetables high in beta carotene do bother me, and I am hypothyroid, temp moved up to 97.8 from taking thyroid glandular, but had been anywhere from 96.6 - 97.6 for many years.

 

[SWAlexander]

One very important aspect is Cell apoptosis.

"In this in vitro paradigm, CGNs undergo rapid and synchronous apoptotic cell death within 24 h after removal of serum and lowering of extracellular potassium."

In this paper: Cracking the code of neuronal apoptosis and survival
https://www.nature.com/articles/cddis2015309
"Hypoxic-ischemic (HI) brain injury is a serious insult that results in various degrees of damage to the body, leading to significant neurobehavioral dysfunctions. The immature brain is particularly fragile to oxygen deprivation (OD), termed HI brain damage (HIBD). Following HI stimulation, whether neurons undergo apoptosis or survive is dependent upon the duration that cells are able to maintain homeostasis following the insult. Autophagy is a cell-autonomous survival mechanism that maintains cell homeostasis. It is an essential catabolic process that degrades misfolded proteins and clears excess or damaged organelles in cells by isolating them in double membrane structures, which are then fused with lysosomes."

Will neuronal cells be replaced after apoptosis?
Do dead neurons get replaced?
Nerve cells in your brain, do not renew themselves.

What are the consequences if nerve cells in your brain, also called neurons, do not renew themselves?
Neurons in the adult, especially in the cerebral cortex, are traditionally thought not to regenerate after maturity, unlike other cell types such as skin cells, liver cells, or blood cells. This means that the neurons you have as an adult are generally the same ones you were born with. When they're damaged or die, they don't undergo mitosis or conventional cell division like some other cells.
Here are the consequences of this non-renewal characteristic:

1. Irreversible Damage: Once neurons are lost due to injury, disease, or aging, they are generally not replaced. For example, neurons lost due to a stroke or traumatic brain injury do not regenerate, leading to permanent deficits.
2. Vulnerability to Degenerative Diseases: Diseases like Parkinson's and Alzheimer's involve the loss or dysfunction of specific groups of neurons. Because these neurons don't regenerate, these conditions are progressive and currently incurable.
3. Dependency on Neural Maintenance: Neurons are long-lived cells. To ensure their functionality throughout life, there's a strong reliance on maintenance mechanisms, including clearing out of waste products and repair of cellular components. If these maintenance processes fail, it could lead to neural dysfunction or disease.

The inability of most neurons to regenerate is a fundamental aspect of the nervous system's architecture that has both advantages and disadvantages. It provides stability to neural circuits, but it also makes the brain vulnerable to injury and disease.

 

[Wishful]

I guess a third possibility is the initial trigger has damaged the immune system in some way.

I think in engineering terms, and the immune system looks like a set of feedback loops. Some feedback is positive, to provide an overwhelming response to an infection, and some feedback is negative, to calm things down after the infection is gone. With a complex set of feedback loops, it's possible for some loops to get locked in one state. That's my favourite theory for ME's root cause. It fits the rapid switching of state that I've observed, and the persistence of the ME state after triggering. In the early stages, it can switch out of that state by numerous factors, but over time, the system adapts to that state, providing more feedback maintaining it, making it harder to switch. That also explains ME's habit of having a treatment work for the first few doses, but then stop working and never working again.

The immune system isn't damaged, it's just reprogrammed to maintain a different homeostatic state.

 

[SWAlexander]

isn't damaged, it's just reprogrammed

That´s the problem. One cytokine storm after another. https://swaresearch.blogspot.com/2023/08/what-is-cytokine-storm.html

I wonder if there is a possibility of damaged neuronal mRNA apoptosis.
Any thoughts or links?
All I found so far:

Neuron-Specific mRNA Complexity Responses during Hippocampal Apoptosis after Traumatic Brain Injury
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6729833/​

 

[Wishful]

My last temporary remission was in year 8 or maybe 10 in my ME, and it still switched to 100% non-ME over the space of minutes, so it pretty much can't be long-term damage. A system of amplifiers with feedback loops can lock into a state indefinitely, but with the right external "poke", it could switch back to its normal state immediately. If the feedback loops are still biased towards that positive feedback loop, it will lock back in after the external "poke" is gone. It's a simple and logical theory for ME, but finding the problematic feedback loop is as difficult as if those electronic amplifiers were buried deep in epoxy, wires and other components, and you weren't allowed to damage it.

 

[SWAlexander]

A system of amplifiers

What could the amplifier be? Any guess?

feedback loop

Would you describe it as an energy boost?

 

[Hip]

Interesting. Dysregulation of TRP melastatin subfamily members (TRPM) and calcium signalling processes are implicated as being involved. In this earlier review here for example, they presented TRPM7 as a potential candidate in the pathomechanism of ME/CFS (one part), as TRPM7 is increasingly recognized as a key mediator of physiological and pathophysiological mechanisms affecting neurological, immunological, cardiovascular, and metabolic processes inherent in ME/CFS. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8535478/

If you are quoting studies, it is normal to use a quote box,

like this.

This is in order to differentiate your own comments from the text quoted the study. You are jumbling both your own words and quotes from the study in the same sentences and paragraphs.

 

[Violeta]

Findings from this investigation suggest that TRPM7-dependent Ca2+ influx is reduced with agonism and increased with antagonism in ME/CFS patients relative to the age- and sex-matched HCs.

Isn't strange that agonism reduces influx and antagonism increases influx? I'm having difficulty understanding this.

 

[Hip]

I just saw that Abilify modulates the ion channel transient receptor potential melastatin 7 (TRPM7).

This might explains why Abilify offers benefits for ME/CFS.

 

[Violeta]

"Subsequent flow cytometric, patch-clamp electrophysiology, and microscopy investigations revealed a loss of TRPM3 ion channel function, reduced surface expression, and impaired Ca2+ mobilisation in the NK cells of ME/CFS patients compared with healthy controls (HCs) "[19,20,21,23,24,25].

Zinc is important for TRPM3, too.

TRPM3 in Brain (Patho)Physiology​

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7959729/

I have to find this quote from the study in the study (I just copied it from the google description of the study.) The study is so intense, I hope no one minds me putting it here.

Zinc plays an important role in the central nervous system as well, by regulating excitability of ion channels and it can be released together...

 

[SWAlexander]

I'm having difficulty understanding this.

I´m not sure if I can explain this correctly.

In ME/CFS (Myalgic Encephalomyelitis/Chronic Fatigue Syndrome) patients as compared to healthy controls who are matched by age and sex.

1. TRPM7 is known to be involved in the regulation of calcium (Ca2+) influx, which is crucial for various cellular functions. Anomalies in calcium signaling can affect cell functions and have been linked to various diseases.
2. When TRPM7 is activated (agonism), there's a reduction in the Ca2+ influx in ME/CFS patients compared to healthy controls. Conversely, when TRPM7 is inhibited (antagonism), there's an increase in Ca2+ influx in ME/CFS patients relative to healthy controls.
3. This suggests that ME/CFS patients have a distinct TRPM7-related calcium signaling pathway response when compared to healthy controls. These differences in calcium regulation might contribute to the pathology of ME/CFS, potentially offering a therapeutic target or a biomarker for the disease.
   
   there are several different test methods:
   Gene Expression RT-PCR (Reverse Transcriptase-Polymerase Chain Reaction)
   Protein Expression/Western Blot. I don´t know anything about it.
   Immunohistochemistry (IHC) or Immunocytochemistry (ICC)
   Calcium Imaging and some more.
   I´m not sure what lab is equipped to do these tests and if insurance would pay for them.
   
   

 

[Violeta]

I´m not sure if I can explain this correctly.

In ME/CFS (Myalgic Encephalomyelitis/Chronic Fatigue Syndrome) patients as compared to healthy controls who are matched by age and sex.

1. TRPM7 is known to be involved in the regulation of calcium (Ca2+) influx, which is crucial for various cellular functions. Anomalies in calcium signaling can affect cell functions and have been linked to various diseases.
2. When TRPM7 is activated (agonism), there's a reduction in the Ca2+ influx in ME/CFS patients compared to healthy controls. Conversely, when TRPM7 is inhibited (antagonism), there's an increase in Ca2+ influx in ME/CFS patients relative to healthy controls.
3. This suggests that ME/CFS patients have a distinct TRPM7-related calcium signaling pathway response when compared to healthy controls. These differences in calcium regulation might contribute to the pathology of ME/CFS, potentially offering a therapeutic target or a biomarker for the disease.
   
   there are several different test methods:
   Gene Expression RT-PCR (Reverse Transcriptase-Polymerase Chain Reaction)
   Protein Expression/Western Blot. I don´t know anything about it.
   Immunohistochemistry (IHC) or Immunocytochemistry (ICC)
   Calcium Imaging and some more.
   I´m not sure what lab is equipped to do these tests and if insurance would pay for them.
   

Okay, that makes it make sense.

And what if that increased influx of Ca2+ is caused by candida albicans.

In which case one would not want to antagonize it.

https://www.biorxiv.org/content/10.1101/2023.08.26.554944v1.full

TRPM7 regulates phagocytosis and clearance of Candida albicans​

 

[Violeta]

Okay, that makes it make sense.

And what if that increased influx of Ca2+ is caused by candida albicans.

In which case one would not want to antagonize it.

https://www.biorxiv.org/content/10.1101/2023.08.26.554944v1.full

TRPM7 regulates phagocytosis and clearance of Candida albicans​

Although maybe it has more to do with a zinc deficiency.

Well, maybe since candida causes a zinc deficiency, it is the cause.

Or maybe since candida sequesters zinc, the NK engulf the candida because they are bound to zinc.

Yes, that's how my brain feels right now.