Oocyte activation and latent HIV-1 reactivation: AMPK as a common mechanism linking the beginnings of all human life and the potential eradication of HIV-1

Source: http://med-ukraine.com/ivf-in-vitro-fertilisation/; https://s-media-cache-ak0.pinimg.com/736x/b6/c3/de/b6c3def9ec8447a5af80f3a0fb690a69.jpg; http://www.shutterstock.com/video/clip-15185026-stock-footage-medical-d-animation-of-a-fetus-week.html?src=rel/5574932:1/gg 

Hey Blogger friends, here's a link to a journal article that I finally got published (after half a year) and it definitely makes some novel and bold observations that affect all of us. Basically, the article says that the process that initiates the beginning and creation of all human life, which is marked by the activation of an oocyte (i.e. egg), is nearly identical to the process that activates a T cell that is infected with dormant HIV-1.  In fact, compounds that have been used to effectively reactivate dormant HIV-1, making it visible to the immune system so the virus can be destroyed, are the exact same compounds that have been used to artificially activate an oocyte during in vitro fertilization procedures, producing live healthy children.  Also, when a sperm activates an egg, the activation of the egg involves the exact same proteins as when a T cell becomes activated, in essence bringing both cells "back to life".  In HIV cure research, this is called the "shock and kill" approach. In my publication, I coin the term "shock and live" approach to describe the analogous process in creating life.  What causes the "shock" in both oocytes and T cells is an increase in calcium levels induced by the sperm or induced when a T cell becomes activated when it encounters a foreign invader.  This "shock" that leads to the creation of life or the activation of T cells can also be induced by heat stress, electrical pulses, and other stressors.  The "shock" induced by calcium activates the master metabolic regulator AMPK in oocytes (novel) and T cells.  AMPK activation is critical for the production of the energy powerhouses of your cells called mitochondria.  Mitochondria are indispensable for both T cell activation and oocyte activation. Basically, AMPK provides that initial "burst of energy" in the form of ATP that is largely responsible for activating an oocyte and a T cell.  This "shock and live" approach is indeed a fundamental theme that pervades not only our physiological bodies but also impacts us psychologically/metaphysically.

"What doesn't kill you makes you stronger" is just a rewording of the "shock and live" approach that is responsible for the initiation of all human life.  Continual challenges, or "stress", to the body leads to a hormetic effect in which the body reacts by becoming stronger, faster, and more agile (i.e. exercise).  These changes are critically dependent on AMPK, mitochondria, and ATP production.  Continual intellectual challenges (i.e. learning a new language, etc) leads to a hormetic effect in which the brain becomes more complex and efficient by producing more neurons and forming more intricate connections in the brain, a process that is again critically dependent on AMPK, mitochondria, and ATP production.  Lastly, extreme hardships experienced in life (e.g. loss of job, divorce, financial strain) are metaphysical manifestations of the "shock and live" approach.  If these experiences don't kill you or permanently strip you of your drive and desire for a fulfilling and successful life, they can be used as a potent fuel source to ignite a fire that may drive you to heights in life that were previously unimaginable or even impossible without those experiences.  Indeed, there are without a doubt bewilderingly successful people who have untold experiences that have darkened many days of their lives.  Their success is, in my opinion, heavily dependent on the application of the "shock and live" approach.  Interestingly, many of the plant-based compounds that have been shown to extend lifespan and healthspan (e.g. curcumin, resveratrol, metformin) are produced by plants as toxins that "shock" small predators to death. However, for humans, this small "shock", on a cellular level, is just the right amount of "shock" to cause all cells exposed to the compound to do what the cell was designed to do that much better. In closing, the very creation of all human life is predicated on a positive and beneficial response of an oocyte to a stressor or "shock", activating the oocyte and giving rise to new life. This response to a "shock" is also what brings a T cell to life to vigorously fight off foreign invaders. This "shock", if voluntarily and continually induced throughout life, will also extend and maximize your life and health, on both a physical and metaphysical level. 

Oocyte activation and latent HIV-1 reactivation: AMPK as a common mechanism linking the beginnings of human life and the potential eradication of HIV-1

*Author's Note: 

In the above referenced recent publication, I highlight the novel observation that the intracellular mechanisms that drive the beginnings of all human life, a point defined by sperm-induced or artificial activation of an oocyte, are strikingly similar to the intracellular mechanisms that drive the activation of T cells and hence the reactivation of latent HIV-1 that lay dormant in memory T cells.

In an active area of research known as HIV-1 cure research, the purposeful reactivation of latent HIV-1 in memory T cells to alert the immune system to the presence of the virus is known as the “shock and kill" approach. As this approach nearly parallels the intracellular signaling mechanisms that lead to oocyte activation, the activation of an oocyte can also be appropriately described as the “shock and live” approach, a term first used to characterize oocyte activation in the above referenced publication.

Indeed, as explained in the Abstract reprinted below, the activation of both oocytes and T cells is critically dependent on an increase in the levels of intracellular calcium (Ca2+). This increase in intracellular Ca2+, which is characterized by waves or oscillations, is induced by the action of different isoforms of the protein PLC (PLCζ in sperm and PLCγ1 in T cells), leading to a signaling cascade involving the same proteins in both T cells and oocytes (e.g. PIP2, DAG, IP3, PKC) that ultimately lead to oocyte activation and T cell activation-induced reactivation of latent HIV-1.

Interestingly, correct functioning of the primary energy-generating organelle of the cell, the mitochondrion, is absolutely indispensable for both T cell activation and oocyte activation, as mitochondria provide the necessary ATP production and Ca2+ buffering capacity essential for activation of both cell types.

This similarity in “bringing back to life” both oocytes and T cells points convincingly to the novel observation that the activation of AMPK, the master regulator of cellular metabolism, represents the central node that characterizes the beginning of all human life via oocyte activation and the potential eradication of HIV-1 through T cell activation-induced latent HIV-1 reactivation.

Indeed, before becoming fully mature and released for fertilization on ovulation, oocytes were once “alive” and had to undergo a maturation process (i.e. oocyte maturation) initiated by a surge in luteinizing hormone before becoming dormant again.  Strikingly, oocyte maturation has been shown to be critically dependent on and induced by AMPK activation and even the production and release of luteinizing hormone itself from pituitary cells is dependent on AMPK activation induced by Ca2+.

Similarly, memory T cells that are latently infected with HIV-1 were once “alive” when initially infected by HIV-1 but later transitioned to a dormant state, with the integrated HIV-1 virus also becoming latent along with the dormant memory T cell.

AMPK is an evolutionarily conserved protein that is present in nearly all eukaryotes and is activated by a variety of cellular stressors that a cell may encounter, including heat stress, an imbalance in the amount of reactive oxygen species production (i.e. oxidative stress), a decrease in cellular energy levels (AMP/ATP ratio), an increase in intracellular Ca2+ concentrations, osmotic stress, aerobic exercise, etc.  The activation of AMPK by these cellular stressors leads to an improvement in functionality and birth of new mitochondria as well as an increase in the levels of enzymatic and non-enzymatic antioxidants that further provide cellular protection.  Interestingly, as studies indicate that mitochondria may have played a deciding role in eukaryotic evolution and the development of complex organisms including humans, it is likely that AMPK was present early during eukaryotic evolution, functioning as a critical energy gauge and inducing mitochondrial biogenesis on cellular stress induction.

Indeed, AMPK activation in lower animal model organisms, including yeast, worms, flies, and mice, significantly improves or extends lifespan and health span. Compounds that have also been shown to improve/extend lifespan and health span in those organisms, including rapamycin, metformin, and resveratrol have each been shown to significantly induce AMPK activation in vivo.

In fact, metformin, a prototypical AMPK activator, has been shown to improve pregnancy rates observed during human in vitro fertilization procedures, beneficially alter gene splicing, and induce the production of CD8+ memory T cells, a T cell subset that is essential for the elimination of both cancer cells as well as viral pathogens.

Moreover, PMA (a PKC activator) and ionomycin (a compound that increases intracellular Ca2+ concentrations) when combined are extremely efficient in reactivating latent HIV-1 via T cell activation (they are used as positive controls) and have also been shown to induce artificial oocyte activation. Indeed, ionomycin has been successfully used to induce human oocyte activation, facilitating fertilization during intra-cytoplasmic sperm injection (ICSI) procedures.  Interestingly, as this publication details, activation of mouse oocytes (models for human oocytes) can be achieved by the application of carvacrol, a compound derived from oregano that activates receptors on oocytes that detect changes in temperature.

Indeed, a novel confluence among lifespan and healthspan extension and improvement, T cell activation-induced latent HIV-1 reactivation, and oocyte activation is also outlined in this publication. As noted above, metformin has been shown to improve lifespan and healthspan in several animal models as well as beneficially alter splicing of the insulin receptor gene in normal human diabetics.  AMPK activation has also been shown to beneficially alter gene splicing in cells derived from patients with the genetic disorder myotonic dystrophy type I, a disorder characterized by aberrant gene splicing.

Surprisingly, several compounds that have been shown to significantly improve accelerated aging defects in the faulty gene splicing disease Hutchinson-Gilford progeria syndrome (HGPS) have also been shown to activate AMPK in vivo.  For example, rapamycin has been shown to significantly improve HGPS cellular defects through the induction of autophagy, a process that involves recycling of worn cellular components. Autophagy is also induced during both T cell activation and fertilization. Methylene blue has been shown to improve HGPS cellular defects by increasing PGC-1alpha-induced mitochondrial biogenesis but also induces oocyte meiotic maturation as well.  Retinoic acid has been shown to reduce progerin mRNA transcript levels in HGPS fibroblasts, alter the activities of the splicing factors ASF/SF2 and SRp55 (splicing factors that are instrumental in orchestrating alternative splicing of the HIV-1 genome and the LMNA gene in HGPS), facilitate T cell activation, and enhance both mouse and human oocyte maturation and activation.  Lastly, vitamin D, via activation of the vitamin D receptor, has recently been shown to correct cellular aging defects in HGPS, reactivate latent HIV-1 in certain cell types, and promote the activation of T cells. 

As each of these compounds activate AMPK in vivo and the deletion or knock down of AMPK leads to T cell death as well as a significant inhibition of oocyte maturation (a process that precedes and is essential for oocyte activation), AMPK activation appears to represent a molecular switch that provides the necessary burst of energy that defines the beginning of all human life and the “bringing back to life” of memory T cells via T cell activation (see Figure below).  That the potential eradication of a disease that has claimed the lives of so many can be realized simply by admiring the process through which all human beings are created is nothing short of astounding. 

http://www.sciencedirect.com/science/article/pii/S0306987716301165

https://www.linkedin.com/pulse/oocyte-activation-latent-hiv-1-reactivation-ampk-common-finley?published=u

Abstract

In all mammalian species studied to date, the initiation of oocyte activation is orchestrated through alterations in intracellular calcium (Ca2+) signaling. Upon sperm binding to the oocyte plasma membrane, a sperm-associated phospholipase C (PLC) isoform, PLC zeta (PLCζ), is released into the oocyte cytoplasm. PLCζ hydrolyzes phosphatidylinositol 4,5-bisphosphate (PIP2) to produce diacylglycerol (DAG), which activates protein kinase C (PKC), and inositol 1,4,5-trisphosphate (IP3), which induces the release of Ca2+ from endoplasmic reticulum (ER) Ca2+ stores. Subsequent Ca2+ oscillations are generated that drive oocyte activation to completion. Ca2+ ionophores such as ionomycin have been successfully used to induce artificial human oocyte activation, facilitating fertilization during intra-cytoplasmic sperm injection (ICSI) procedures. Early studies have also demonstrated that the PKC activator phorbol 12-myristate 13-acetate (PMA) acts synergistically with Ca2+ ionophores to induce parthenogenetic activation of mouse oocytes. Interestingly, the Ca2+-induced signaling cascade characterizing sperm or chemically-induced oocyte activation, i.e. the “shock and live” approach, bears a striking resemblance to the reactivation of latently infected HIV-1 viral reservoirs via the so called “shock and kill” approach, a method currently being pursued to eradicate HIV-1 from infected individuals. PMA and ionomycin combined, used as positive controls in HIV-1 latency reversal studies, have been shown to be extremely efficient in reactivating latent HIV-1 in CD4+ memory T cells by inducing T cell activation. Similar to oocyte activation, T cell activation by PMA and ionomycin induces an increase in intracellular Ca2+ concentrations and activation of DAG, PKC, and downstream Ca2+-dependent signaling pathways necessary for proviral transcription. Interestingly, AMPK, a master regulator of cell metabolism that is activated thorough the induction of cellular stress (e.g. increase in Ca2+ concentration, reactive oxygen species generation, increase in AMP/ATP ratio) is essential for oocyte maturation, T cell activation, and mitochondrial function. In addition to the AMPK kinase LKB1, CaMKK2, a Ca2+/calmodulin-dependent kinase that also activates AMPK, is present in and activated on T cell activation and is also present in mouse oocytes and persists until the zygote and two-cell stages. It is our hypothesis that AMPK activation represents a central node linking T cell activation-induced latent HIV-1 reactivation and both physiological and artificial oocyte activation. We further propose the novel observation that various compounds that have been shown to reactivate latent HIV-1 (e.g. PMA, ionomycin, metformin, bryostatin, resveratrol, etc.) or activate oocytes (PMA, ionomycin, ethanol, puromycin, etc.) either alone or in combination likely do so via stress-induced activation of AMPK.

Keywords:

AMP-activated protein kinase (AMPK); Oocyte; HIV-1 latency;Hutchinson–Gilford progeria syndrome (HGPS); LKB1; CaMKK2; Phospholipase C zeta (PLCζ); Phospholipase C-gamma 1 (PLCγ1); Bryostatin; Resveratrol; Metformin; Retinoic acid