Pillar 2: Detoxification module 2 3

Hello and welcome back. This is pillar 2 module 2.3 and we are focusing on the glutathione S transferase enzymes. We'll do a quick review of phase two detoxification. Talk about Glutathione Transferase SNPs, specifically GSTP1, as well as the common glutathione transferase deletions. We have already talked about phase one detoxification and this includes the cytochrome P450 genes that code for those phase one enzymes, also referred to as the cyps. The cyps. So we have fat soluble chemicals and toxins or endogenous compounds, hormones. These are converted by phase one enzymes, the cytochrome P450s or CYP450s. And then we have these intermediate metabolites. So after phase one, the compounds that are produced from this first step are often more reactive. These can cause oxidative stress. These are often oxidative molecules or unbalanced reactive compounds that can damage DNA and cellular health. So, so what your, you know, endocrine system, whatever it may be. So we're really getting to these underlying triggers and pathways that we want to understand and we want to support so that that person can get back to homeostasis or at least identify it. Even if they're coming to you and they're not at this state, which is ideal, they're coming to you for healthy aging or just understanding their unique genetic differences. You can guide them on whether they are more likely to be sensitive to chemicals and toxins and what they can do because there's so much we can do to reduce our toxic load and very quickly. These glutathione S transferases, the family, are classified into alpha, mu, PI and theta. So you'll see GSTM or GSTP or gstt if it's theta. I won't go into the classification, but we did talk about it in the cytochrome P450s. And just remember that these classifications are based on similarities in amino acid sequence and structure. To put it simply, the sole function of the glutathione S transferase enzymes are to drive this reaction or this conjugation, the bringing together of glutathione and a xenobiotic. As you can see at the top of this diagram, that xenobiotic being either an environmental pollutant chemical, toxin, endogenous reactive compound. The coming together after this GST glutathione enzyme reaction is a resultant glutathione S conjugate molecule. This new glutathione conjugate is no longer as reactive and potentially dangerous. It's not going to cause that DNA damage, it impact cellular health or increase oxidative stress. It's now water soluble, neutralised and can be excreted from the body. If you'd like to know more about the glutathione S transferases as a whole, these family of phase two detoxifications and their impact on health and chronic conditions and disease, you are welcome to check this paper. As I said, the textbook doesn't have a lot on these particular genes and genetic variations. So the the paper is added into this module if you want some more information and get stuck into the science. Glutathione S Transferase P1 or GSTP1 is located in the breast, predominantly in the breast, brain, lungs, the colon, the kidneys and the skin. And this distribution pattern suggests that GSTP1 has a special role in protecting tissues that are away from the liver. A lot of these GSTs are are found within the liver, predominantly in the liver. And we know that is the important organ when it comes to detoxification. But we also need to clear chemicals and toxins from other tissues in the body like the breast, the brain and the lungs. And this is where GSTP comes in. And if you're wondering why it's gstp, remember at the beginning we talked about the different classifications, PI theta mu, so it can be GSTP1 referring to PI Mu or Theta T1. So within GSTP1 you want to look for the genetic variation that causes an amino acid change, isoleucine to valine at codon or amino acid position 105 in that amino acid chain, the nucleotide or that base pair change is an A to a G at position 3 1, 3 in that nucleotide sequence. But most of the time in the published literature or if you're looking at a DNA report, they'll often refer to that amino acid change. Just like with comt, we tend to refer to the amino acid change rather than that nucleotide or base pair position. Individuals who inherit the G that variant or valine allele, exhibit lower enzyme activity and potentially have a reduced ability or capacity for detoxification of particular chemicals and toxins within these tissues where GST is predominantly located. So if you are looking at results in your report, the aa, which is the isoleucine isoleucine that is associated with normal enzyme activity, we then have intermediate for the heterozygotes, the A and the G, or those that inherit the homozygote. So A double variant, a gg, which in the amino acid sequence would be a valine. And a valine. This changes that structure of the enzyme. It doesn't function as effectively and that's associated with reduced detoxification capacity. Therefore, those that have the GG genotype may have increased susceptibility to detoxification of certain chemicals and potentially a buildup of oxidative stress. They don't have that same antioxidant capacity. They can be sensitive to certain chemicals and toxins, including estrogen metabolites in the breast tissue. These SNPs or this particular SNP has been linked to cancers, breast cancer, lung cancer, bladder cancer. And that's no surprise because it's predominantly found in these tissues and associated with clearing chemicals and toxins that can trigger cancer and DNA damage. In addition, this SNP has been linked with respiratory disorders, specifically asthma, and sensitivities to chemicals and toxins that are breathed in and tobacco smoke. And there is some evidence that this particular SNP can alter the response to chemotherapy. So that is amongst a number of the GSTs, actually. So if you are involved in integrative cancer therapy, you can look into this. So I have one research paper here. If you are working in the space of cancer, you can review this. What I want to say, though, is at this point in time, you know, in the middle of 2025, although we have a lot of knowledge and there does seem to be potentially higher risks or sensitivities to some of these cancer medications, with genetics, we still have a lot to learn. So you need to be cautious with the language. And if you are not working in the space of cancer therapy, you're not an integrative oncologist or you're not a specialist in this area. This is outside your scope and you are not recommended to talk about GSTP1 and cancer therapy, particularly if the person doesn't have cancer. And it's not your area of expertise. So remember, you're practicing within your scope. What you would talk about is the impact of GSTP1 and that's influence on chemicals and toxins. And, you know, if they've got asthma or they are having signs and symptoms linked with exposure to certain chemicals and toxins, you can help them understand in their body why they might be a bit more sensitive. But you're not going to talk to them about potential risks of cancer treatments and therapies. You leave that to the experts. I do imagine in the coming years with AI and the information that's coming through around genetics and this Being such a hot topic and research dollars being invested into it. There will be, I imagine, the ability to use AI, large data sets when we potentially have the whole genome. And we will be able to identify who might be better suited to certain, you know, medications, cancer treatments, things like that. I mean, it's already happening in a small degree, but we have a long way to go when AI does come into place. And we do have these tools that are robust and reliable and helping people with this personalized medicine. Your role as an integrative practitioner or allied health, you know, functional medicine, doctor, whatever it may be, is never going to be redundant because people are always going to be looking for experts to explain this. Someone's going to want to understand what do these genes mean? Why have I been told this? And also they need people like us to help train these algorithms and things that actually understand the genes, the interactions with chemicals and toxins, how that influences health. So this is in your portal. If you are interested in this space, you can check this paper. This study has just been published June 2025, and it's looking at the risk of cardiovascular complications among type 2 diabetes patients with the GSTP1 SNP. So typically, if you are very much focused on cardiovascular disease or type 2 diabetes, you may go to other SNPs, TCF7L2 or others that are influencing cholesterol or homocysteine that are linked with cardiovascular disease. These are genes and genetic variations that have a direct link. For example, if we're thinking diabetes, you might be looking at SNPs that influence directly influence blood sugars. But with GSTP1, we know there's an indirect influence because GSTP1 impacts levels of oxidative stress as well as the clearance of chemicals and toxins, endocrine disruptors, things that can influence cellular health, epigenetics, things like that. So there is this indirect link. So it's no surprise that there may be an association because of that oxidative stress, cellular dysfunction. But it's not a direct link in association with, say, you know, blood sugar regulation and diabetes or some of those, you know, direct causes of cardiovascular health. So that's the first thing I want to say. Secondly, I've chosen this paper to show you one. I was excited to see a brand new paper, but also this is looking at two different SNPs within GSTP1. So at this stage, I've just gone through one SNP with you, the RS1695 that's looking at a genetic variation at position 105 in that amino acid sequence. So in that protein at position 105, we have an isoleucine, which could be a valine, and those that have the valine, but that change we know that influences the enzyme's ability to clear chemicals and toxins. They have identified another SNP or tested for another SNP a little bit further down in the protein at position115.115, where there's an alanine to valine variation or they call a mutation. But not only is this looking for associations with cardiovascular risk in these patients with diabetes, they've done some functional analysis or sort of looking at the mechanism, and they've done some docking studies and found that a docking analysis revealed that the 105 variation, which I've focused on so far, plays a significant role in altering GSTP1 capacity compared to the alanine 115 variation, or mutation, as they call it. So I'm reading this bottom text box here, and this suggests that the 105 mutation has a greater impact on the protein's short structure, function or susceptibility to disease compared to this variation. A little bit further down. 115. And alanine to valine. So, in summary, the authors indicate that genetic alteration in GSTP1Rs 1695, which is that 105 mutation, or what we call a SNP, can contribute to risk of type 2 diabetes and cardiovascular disease. And as I said, if that is the case, it's likely to be through that mechanism of oxidative stress. So although they have tested another SNP and it's likely that they have chosen this secondary SNP or this other snp, because another paper perhaps has shown there's an association, but from a functional impact, from a mechanistic perspective, it doesn't seem to change the protein structure to the degree of the SNP I've told you about, and therefore doesn't influence the function or the susceptibility to disease to the level that the RS1695 does. Why is this important? Why am I discussing it? It's because when you are reviewing DNA test reports, you'll often see other SNPs and you may think, wow, I haven't heard about this one. You know, why is that here? And it's likely that there has been some publications or evidence to suggest there's an association, but often you. And I cannot say this for sure, but it's highly likely that those other SNPs will not have the level of evidence or such a strong functional effect as the SNPs that I'm highlighting in the academy. Now, that doesn't mean you're going to learn about every single SNP in the academy. Eventually we'll keep updating and updating, but the ones that I believe are the heavy hitters, which 3x4 calls those pop stars and heavy hitters and pop stars, these are obviously not scientific terminology. But those that have a large functional effect we have seen over and over in different studies from cell culture work, animal work, human studies, we're very confident that they have an influence on that pathway or system and they're linked to a health outcome. And there's obviously something that we can do about it. They're the genes we want to focus on. And then we can be looking at the other peripheral genes that potentially have a weaker association. And I say this, and I know I say it all the time, but I'm reminding you, if you have a report and someone says, oh, we've got 600 SNPs, it's not necessarily better because it may be that 400 of those either have very weak evidence or in some cases they haven't had the people on board to necessarily do the right research. Maybe there was one genome wide study that showed one association, but nothing has backed that up. We don't understand even what the gene does or the mechanism. So in this case we understand the gene, we understand the mechanism, but the SNP doesn't have the same functional effect as that. Rs1695 My suggestion to you, as I've said in the mentoring calls, and I say over and over, is you can contact the testing company if they bring out a new report and there's new SNPs or you hear about a new company and you want to get that DNA report. I do these things all the time. I'm always interested in what's coming about and how they explain the results. And is this something better that I could use with my patients or clients or is it going to help me understand my health or my body? Is there more in this compared to what I've seen in the past? Is this going to help me level up even further? But you will come across some new snips and then the quickest way to find out if these are really, you know, evidence based, heavy hitting snips like the pop stars that have a big functional effect is to contact that company and say, I'm not familiar with this snip, can you please pass on the research or you know, let me know why you decided to include it. Of course you can do your own research. I love doing research, I love learning, but I don't have the time. If I had days and weeks, I, I would just sit down and look into all of these new SNPs, but I don't have that time, so. And most practitioners are very busy. You know, you're doing this course to learn what you need to get set up in clinic. But these sort of secondary SNPs or those that may not have such a large effect, rather than getting lost in the research, you know, the best way is to reach out to that company. They've obviously spent the time, or should have spent the time researching, and then they can tell you directly why it's been included into the report. Okay, last study. So, very quickly, the reason I wanted to highlight this, there is a lot of studies with GSTP1 suggesting that it's linked with breast cancer. Remember, it's located in the breast and it's involved in clearing those estrogen metabolites. But what I wanted to highlight here is this paper is a little bit strong in its language. It says down the bottom in the orange square that the GSTP1 105 isolucentivaline that I have gone through with you, and they're also showing that nucleotide, the base pair sequence. So at 313 in the base pair on nucleotides, there's an A to a G. So we're seeing here the amino acid and the nucleotide. Just to make that clear, for those of you who understand this completely, I'm reinforcing these things for the new people. And if you still don't understand this, please ask me in the mentoring sessions. But what they've claimed is that this SNP is an independent or considered an independent risk factor for breast cancer in Egyptian women. None of these genes with common SNPs, like Rs1695 that we're talking about are independent risk factors. So we're looking at functional genomics. Nutrigenomics is the interaction with our diet, key nutrients, environmental factors. So just be cautious. I could go into a whole big story here about in the 90s and early 2000s, when some genes, potentially even MTHFR and homocysteine and all of these things were once upon a time thought independent risk factors for cardiovascular disease, et cetera. But as time's gone on, we've realised they're not. In some people, they don't cause disease. In some cases with homocysteine, we now think Maybe it even comes after someone's having some of these other cardiovascular risk markers. So they're not independent, they are associations potentially. We could use words like susceptibility, but predisposition to breast cancer, but not independent risk factor. So just be mindful of that. And when you're going through the research and learning more, we're always talking about the interaction between these SNPs and the environment and then that is what is linked with either health or disease. Alright, in the previous module we talked about our cytochrome P450s, our phase one detoxification enzymes, and now we're onto our phase two. And if we are thinking about our hormone metabolism, we can see on the left here we've got estradiol being converted into potentially those semiquinones which specifically happens, or is more likely to happen with that cytochrome P451B1 that pushes down that 4 hydroxy, which can then lead to these semiquinones that are very reactive, but not as reactive as the quinone. So if you can see semiquinone in the middle of this slide, you can see it goes to quinones. So quinones are very reactive. They're what cause DNA damage. They're that sort of link, that mechanistic step into cancers and, you know, serious cellular damage and dysfunction. So these quinones, if estrogen goes down this pathway, or benzenes that turn into quinones or other chemicals and toxins, we have our glutathione transferases, as we've been discussing, that can help clear these reactive, potentially dangerous compounds in the body. But it's not GSTP1 that. It's not only GSTP1 that can help clear these quinones and clear our estrogen metabolites. We also have other glutathione transferases. Remember I said at the beginning of this module, there's a family of them. And many of the testing companies you'll look at will have GST1 and GSTM1 as well. And these are a little bit different because they're not SNPs, they're not a single nucleotide polymorphism. They are shown generally as deletions or null genotypes. So they can be copy number variants and some people can have no copies. And that's generally shown null, means there is no enzyme there, no protein. And then just above that, just to acknowledge the mnsod. So that superoxide dismutase, we'll talk about that in the oxidative stress modules. But that can also help with reducing things like superoxide, which can be produced from these reactive compounds. And superoxide itself is also very dangerous. This is a pro oxidant, a free radical that can also cause DNA damage. So you can see here why these, these enzymes are so important in stepping in and clearing these potentially dangerous chemicals, compounds and molecules. Because if they build up, we can start to get that cellular damage that then influences the whole organ, the whole system. We start to then, you know, particularly, you know, mnsod, you know, in the mitochondria we start to get this buildup of oxidative stress. The mitochondria doesn't function so well. We have that fatigue where then we're not getting the ATP to fuel all these other metabolic reactions as well. So just highlighting those glutathione transferases there because we are now going to get into GST1 and GSTM1 that as I said, are shown in many of the testing reports in relation to detoxification. Similar to GSTP1, GST T1 and GSTM1 are also genes that code for glutathione S transferase enzymes. However, the GSTT1 M1 are predominantly found in liver and kidneys and some other tissues. So where most of our detoxification enzymes are in the liver but also the kidneys. Whereas remember, P1 was, you know, breast and brain and lungs, et cetera. As I just said on the previous slide, we are looking at deletions. So there are copy number variations, not SNPs. And individuals can have one copy, two copies or zero copies. And that, that's referred to as the null genotype or sometimes shown as a deletion, meaning the person does not have the gene, therefore they do not make that enzyme and they do not have that GST one or that GSTM one. And that can impact their detoxification capacity, especially in response to environmental toxins. So I want to make it clear here, if someone doesn't have a high exposure to chemicals and toxins, this is likely to have really no impact. It's more of a concern when someone is working in a job or living in a place that has a high level of chemicals and toxins, this is then putting more pressure on the detoxification system. And potentially there can be this sort of buildup of reactive compounds or yeah, dangerous things in the body if they're not clearing them. I'm jumping ahead here, we've got other slides, but just to clear this up, because sometimes this can be quite scary to people to think, oh, I don't have this really important glutathione transferase enzyme and I may not be able to clear chemicals and toxins. Remember, there is a whole lot family of these glutathione S transferases. There are other enzymes that can step in. And we do have some evidence that when people have these deletions, we are no surprise, more reliant on some of these other glutathione S transferases. So it's not that there's no glutathione conjugation going on at all, they just do not have these specific glutathione transferase enzymes. So in this table, I have tried to summarise a lot of information, but hopefully it is helpful for you in that it includes the main role of these glutathione transferases, key compounds that are detoxified. This is not an extensive list, but some of the common things that you'll see in the literature where these enzymes are predominantly found. So these are not the only tissues, but predominantly found in liver, kidneys, white cells, the genetic variation in this case, we're talking about deletion. So that null genotype and how frequent they are in the population, you will see they're quite, quite common. And then the clinical insight, what this null genotype or deletions means. So therefore no enzyme activity, increased susceptibilities to chemicals and toxins or drug sensitivity, potentially increasing risk of oxidative stress or certain health outcomes that are driven by oxidative stress or driven by DNA damage from these harmful compounds. Really talking about cancers, there's. Just before we move on, I want to highlight that where it says the main role and the key compounds, generally, you don't need to get lost in this specific detail. We don't need to go through specific chemicals and toxins with our clients and patients. What we want to do is just make them aware of the amount of chemicals that they are likely exposed to through their skin care, their household cleaning products, if they're using fragrances, smelly things to make the house nice or the car smell nice, when really that is just putting more chemicals into their system. You know, things that are sprayed on their food. So just bringing this awareness into their lives, if they don't already have it, and then helping them make better decisions, suggesting products and brands that, you know, have a much lower chemical composition, ideally natural things, variations. So looking at this person here in the black T shirt, for example, these little flags across the chromosome there are representing genetic variation. So let's say maybe There's a glutathione deletion, there's something in the GSTP1. There's also genetic variations in other phase two detoxification enzymes influencing a number of these phase two pathways. He's more susceptible to chemicals and toxins. However, he's living this lifestyle. He's smoking, he's drinking, and then many years later, what we see is that he ends up unwell. He hasn't been able to tolerate that lifestyle. Compared to the person on the right, who can see at the top, also looks like they're drinking and smoking, but they've got different genetics. They've been able to tolerate more of this smoking and drinking because they've been able to these chemicals and toxins at a faster rate, so turning them into something that's less harmful and excreting them. So that person isn't as susceptible or predisposed to, for example, in this case, cancer, compared to someone else. Now, of course, there's always a threshold. We're not going to say to our clients, you've got really, you know, this is a strong area. This is one of your genetic strengths, is detoxification. Go out and drink and smoke, because the reality is you will be doing some damage. But some people will tolerate this much better than others. And that is the point of this diagram, just before we finish up. You know, cancer is a genetic disease and I've talked a lot about it. I know not everyone is working in the space of cancer. However, this may have caused a bit of alarm in you as a practitioner. If you have all of these genetic variations or you have some deletions, I want you to understand that with the knowledge you have as an integrative practitioner, you know, naturopath, someone working in the health space, we really can control our environment and our nutrition to reduce our risk significantly. And it's not just cancer, it's all chronic diseases. But specifically in relation to cancer, I wanted to mention that it's not just about the genes you inherit, which, if you're looking at the diagram here on the right, is under hereditary. So you can inherit these genetic variations. Now, there can be strong genetic risks, BRCA1 and 2, for example, or there can be these more latent genetic effects that are very much influenced by our environment, like our GST genetic variations that we've explained in this module. But UV radiation can cause DNA damage, chemicals and toxins, of course, which we've talked so much about. When it comes to detoxification, if you're not getting that heavy load, you're not going to get those. Follow on. Effects of DNA damage, cellular damage, oxidative stress, Viruses can trigger genetic changes, smoking. This really fits into chemicals and toxins, but also when our cells are dividing. So this is an updated image from the National Cancer Institute. I found this when I was looking for that original PowerPoint because I knew I wanted some of those slides. I thought it's a really nice way to explain it with those images. But when it comes to cells dividing, when we get a buildup of DNA damage, like chromosomal anomalies, you know, the micronuclei chromosome breaks, things go wrong at this stage. Often that's due to nutrient deficiencies like what we talked about in methylation, with the importance of folate. When we do cell culture and we take folate out of the media as cells replicate, we get all this DNA damage because it's the folates needed for, you know, the basic building blocks, those nucleotides. So if there are nutrient deficiencies, that is going to cause genetic changes during cell division, but also exposure to chemicals and toxins will influence that as well. So, finally, our last slide. When we are thinking about our susceptibility to chemicals and toxins, it really is a combination of our genes and environment. And this is not just to do with detoxification, by the way. When we are thinking about functional genomics, you know, lifestyle, genetics, this is in all aspects, whether we're thinking about diabetes, whether we're thinking about Alzheimer's, whether we're thinking about optimal performance in relation to, you know, sports performance or cognitive performance, whatever it may be. It is the interaction between the genes and the environment. And when we understand our genes, we can then tailor our lifestyle and our diet to work with our DNA so that we are giving ourselves the best chance to express this really healthy phenotype. You know, phenotype is what you're expressing. Are you expressing high homocysteine? You know, that is not associated with a healthy phenotype. Are you expressing high blood sugars? You know, we can change our phenotype. I'm not talking about classical phenotype like eye color and, you know, natural hair color, though. You can change your hair color but, you know, phenotype here I'm talking about this snapshot of the current state of health that can be used to prevent, diagnose or treat disease or improve health. And this phenotype is influenced by genomics and our lifestyle and our environment. And we will really get into phenotyping in pillar three because we look at the genetics, which is associated with predisposition and susceptibility. But then we're looking at the diet, the lifestyle, complementary testing or blood biomarkers to confirm that phenotype. If those genes are having an impact, and if we really need to step in and try to compensate for these because someone's health is being negatively affected.