FREE shipping on orders over $75!

Your Trusted Brand for Over 35 Years

Funding Scientific Research

The Principal Mission of Life Extension®

Since its inception in 1980, Life Extension (LE) has conducted scientific research that goes beyond the scope of academic institutions and biomedical companies. The purpose of this research is to identify and validate technologies that can delay/reverse aging and prevent premature death.

Mainstream research today is focused on how to treat heart attacks, cancer, Alzheimer’s disease and strokes. These are the diseases that we generally assume cause death.

What most doctors don’t yet recognize is that these devastating illnesses are caused mostly by aging. Life Extension has provided more than $175 million to scientists across the country to look beyond the disease state and instead search for authentic anti-aging and anti-death solutions. Our objective is to prevent or postpone age-related disease, restore health, and provide much longer and higher-quality human life spans.

This annual report will inform you about the research programs we are funding and detail LE’s commitment to meaningful scientific discovery. It outlines how Life Extension continues to fund targeted research into killer diseases such as cancer, cardiovascular disorders, immune dysfunction and neurological deficits. These programs are part of a strategic vision to limit or prevent diseases as we mature.

LE funds researchers denied funding by governments

Accelerating stem cell research

As described in the January 2014 and May 2015 issues of Life Extension Magazine®, Life Extension has been funding many researchers who have been denied funding by the federal government. The following research teams have received support from Life Extension:

Vera Gorbunova, Ph.D., and Andrei Seluanov, Ph.D., are wife-and-husband professors at the University of Rochester in the state of New York. The couple's research has been funded by Life Extension (profiled in the May 2015 issue of Life Extension Magazine®). Dr. Seluanov has the second-largest naked mole rat colony in the world, which he was in danger of losing had it not been for Life Extension funding. Naked mole rats are about the size of mice, but live at least ten times longer than mice, without evidence of aging1 or cancer.2 Dr. João Pedro de Megalhães from the University of Liverpool, UK, attempted to analyze the naked mole rat genome with the couple, but could not find genes which explained either the longevity or the cancer-resistance.3 The discovery by Dr. Seluanov and Dr. Gorbunova that a large molecular weight hyaluronic acid secreted by the naked mole rats protects the mole rats from cancer, led to a cover story in the prestigious journal NATURE.4 Their subsequent discovery of exceedingly error-free protein synthesis by the naked mole rat5 caused the journal SCIENCE to name the naked mole rat "Vertebrate of the Year" for 2013.6 Precision protein synthesis by naked mole rats contributes to both their cancer-resistance and longevity. The next year the couple co-authored a review of cancer resistance and longevity in many species that was the cover story of the August 2014 issue of NATURE REVIEWS: GENETICS.7 More recently, the couple has compared the genomes of mice, naked mole rats, and humans, finding support for the idea that genome maintenance (DNA repair) is important for longevity and cancer-resistance.8 Dr. Gorbunova has particular expertise in DNA repair. She discovered that the sirtuin SIRT6 more than triples repair of DNA damage.9 With Life Extension funding, she has been searching for molecules which will stimulate SIRT6 activity.

Dr. George Church, Harvard Medical School geneticist, was featured in the May 2013 issue of Life Extension Magazine. This article reported on how the Life Extension Foundation was funding the collection and analysis of genes of supercentenarians (people living to age 110 or older) to discover protective genes that allow them to live so long. This funding was provided to a group called Androcyte LLC that initially consisted of CEO James Clement and his assistant, Parijata Mackey. They traveled the world to collect tissue samples from approximately 60 supercentenarians and their family members. Dr. Church was collaborating with Androcyte to analyze the genes.

Since then, Dr. Church has achieved additional fame as being a co-inventor and pioneer in the new CRISPR gene-editing technology. Also since then, the Life Extension Foundation continued to fund Androcyte to open a laboratory in California dedicated to applying CRISPR to deliver longevity genes, initially to mice. Androcyte CEO James Clement continues to work with Dr. Church in doing this research.

Androcyte currently has a colony of 300 mice, and growing. Sixty of these mice were received from the National Institutes of Aging and are between 26 and 36 months of age—the equivalent of very old humans. Androcyte has targeted about 25 promising longevity genes, which are being tested in the mice via CRISPR/Cas9 gene therapy. Particular attention is being paid to the elderly mice to see if they can be restored to youth and good health. To keep costs low, Androcyte purchased a one-acre property with an existing 1,500 square foot building which is an hour’s drive from Los Angeles. As it outgrew its initial vivarium (housing for mice), it added two office trailers to the property to provide additional vivarium and laboratory space.

In addition to Dr. Church and other expert consultants, Androcyte CEO James Clement has acquired the assistance of two new interns: Ellie Dubrovina and David Falzarao, who were referred by Aubrey de Grey’s SENS Foundation. Ellie assists with the scientific work, whereas David assists with the care of the mice.

Androcyte has also received two elderly Arabian mares 28 and 30 years old (age-equivalent to 80-year-old humans) from a sanctuary. If genes delivered by CRISPR to the mice are able to restore youth and health, CRISPR delivery of those genes will be tested on the horses to show that large animals can also benefit. Success with the horses could pave the way for using CRISPR to bring better health and greater longevity to humans.

Victoria Belancio, Ph.D., is an assistant professor at Tulane Medical School who has been using Life Extension funding to study the molecular biology of retrotransposons ("jumping genes") in chromosomes, and the resulting genomic instability.10 She has determined that increased retrotransposon activity results from light-at-night (often due to shift work) which can cause aging11 and cancer.12,13 She has determined that light-at-night, retrotransposon genetic instability, and melatonin inhibition of retrotransposon activity are relevant for breast cancer risk and breast cancer resistance.13-15 Aside from demonstrating the use of melatonin to inhibit the genetic instability due to retrotransposons, Dr. Belancio has found that an antibody can also be used to reduce retrotransposon-induced genetic instability.16

A startup company that Life Extension has been funding is Advanced Neural Biosciences (ANB). ANB has been studying the mechanisms of damage that occurs to blood circulation in the brain in conditions such as stroke, with the aim of reducing those effects. ANB has developed a mathematical formula that can predict the amount of blood-flow impairment resulting from stroke-like states based on electron micrograph images of the brain. ANB reported that in 2015 the company has developed compounds that can restore brain blood flow after two hours without blood flow. Normally, shorter periods of time without blood flow in the brain prevents any subsequent blood flow.

In 2015, Advanced Neural Biosciences started a comprehensive screening program to identify cryoprotectants that can inhibit ice formation in the brain without causing extreme brain shrinking (as existing vitrification solutions do). They have identified a number of promising directions and have devoted most of their efforts in 2016 to validate a number of these vitrification solutions in an in-situ brain cryopreservation model. This is a very promising discovery in the field of neural cryobiology and most of ANB's efforts will be aimed at further validating the glass-forming and toxicity properties of these agents.

In collaboration with a prominent deep learning researcher at the University of Chicago, and Michael Perry, Ph.D. at the Alcor Life Extension Foundation, ANB has developed a computer algorithm that can be used to infer the duration of cerebral ischemia from a set of electron micrographs. They have completed a first draft of a paper that characterizes the histological changes at various durations of cold and normothermic ischemia, which will also include this pioneering application of deep learning in histology. This paper will be submitted for peer review in 2016 and submitted to a critical care medicine journal.

In collaboration with the Alcor Life Extension Foundation, ANB has completed a comprehensive research project to understand the effects of different classes of medications on the perfusion of the ischemic brain. They have identified a number of specific medications that allow ice-free cryopreservation of the brain up to 2.5 hours of normothermic ischemia if administered prior to the insult, and have also investigated the limits of this approach when these medications are administered after the ischemic insult.

In 2015, ANB's lab signed a 5-year lease to conduct research at a stand-alone facility in NE Portland. This additional lab space and lease security allows them to conduct more research models simultaneously and enhance their current capabilities. In particular, they have started work to create a more advanced in-situ brain cryopreservation perfusion room that allows for automated perfusion and data collection in a temperature-controlled enclosure.

Funding a Former Ellison Foundation Scholar

Former Ellison Scholar James Shorter, Ph.D., (University of Pennsylvania Medical School) continues to develop heat-shock proteins that antagonize the misfolding of the amyloid-beta protein linked to Alzheimer's disease with funding from Life Extension. Dr. Shorter has been identifying the most effective of these heat shock proteins, and has found that many of them are also effective against the misfolding of the tau protein that causes Alzheimer's disease. He is now doing his testing in human neuroblastoma cells rather than yeast. He is also trying to convert misfolded amyloid-beta and tau protein into non-toxic species. If this research is successful, it could lead to the first real treatment for Alzheimer's disease, a condition that is currently irreversible, always leading to increasing dementia and ultimate death. Dr. Shorter has also been working with researchers in Switzerland in using his heat-shock proteins to reverse the protein aggregation that causes Parkinson's disease.

For many years Life Extension has been funding cryobiological research that would allow for increasingly long-term, low-temperature maintenance or storage of transplantable organs. Too often vitally-needed transplantable organs from accident victims do not reach needy patients soon enough because the organs deteriorate so rapidly. The demand for transplantable organs greatly exceeds the supply. Just as refrigerators and freezers preserve food, low-temperatures could preserve transplantable organs. The lower the temperature, the longer the organs could be preserved. But because freezing damages all biological tissues, cryoprotectants (anti-freeze solutions) must be developed to prevent ice formation. 21st Century Medicine (21CM) is the major cryobiological research company that Life Extension has been funding. 21CM has developed a relatively non-toxic cryoprotectant solution. The word "relatively" must be emphasized, because all cryoprotectant solutions have some toxicity. For that reason, cryobiologists try to use just enough cryoprotectant to prevent ice formation, but not enough to damage tissues as a result of toxicity. Unfortunately, the organ that is in greatest demand for transplant, the kidney, has posed serious problems for cryobiologists. The outer layers of the kidney receive high blood flow, whereas the inner layers receive low blood flow. So the challenge has been to get enough cryoprotectant into the kidney from the bloodstream to prevent ice formation in the inner layer without causing toxic damage to the outer layer of the kidney. The year 2016 has seen major technical breakthroughs by 21CM on this problem. 21CM can now substantially reduce or eliminate ice formation in the inner layers of the kidney without toxic damage to the outer layers of the kidney.

Up to 900,000 organ transplants are needed each year in the United States alone, compared to about 30,000 transplants actually carried out each year.  Many exciting advances are leading to ways to eliminate the organ shortage, but an avalanche of new organs will create a new problem – how to manage all of the resulting transplants.  Even without the added stress of a 30-fold increase in the number of transplants, transplantation today is a frenetic endeavor, with Lear jets rushing human hearts to their recipients before their short survival times outside the body expire, surgeons performing transplants in the middle of the night, families constantly on standby, waiting to go to the hospital at a moment’s notice to receive a life-saving organ, and with kidneys and other organs being transplanted with considerable storage damage and with less than perfect matches because the alternative is to discard the poorly matched organs as their viable storage times expire. 

Organ banking at very low temperatures (cryogenic temperatures, such as, more specifically, -100 to -140°C, or about -150 to -220°F) would solve all of these problems, and others, because such temperatures arrest biological time, in principle enabling organs to be used at optimal times, and to be transplanted into optimal recipients, without any rush.  At temperatures in this range, storage times become for all practical purposes unlimited.  So far, however, the overwhelming scientific problems of banking organs at cryogenic temperatures have eluded a solution. 

Fortunately, that situation is now rapidly coming to an end, thanks to the indispensable support for organ banking research uniquely provided by the Life Extension Foundation.  The Life Extension Foundation has been willing to support the ambitious goal of organ banking over long periods of time, and even when progress was slow and setbacks were all too abundant, which is the kind of commitment that is absolutely necessary, but all too rare, for the achievement of any very difficult goal as quickly as possible.  But now, results at 21st Century Medicine, just in the first 8 months of 2016, have finally transformed the prospects for success.

To be banked at low temperatures, about 60% or more of the water in the kidney must be replaced with chemicals that prevent the remaining water from freezing so that the kidney will not experience injury from ice crystals.  Not surprisingly, this replacement of water with chemicals has been very difficult to do without damaging the kidney in the process.  Now, however, it appears that this long-standing problem has finally been overcome. 

When a kidney is transplanted, a common way to measure how damaged it is is to follow blood levels of a chemical called creatinine, which is removed from the blood quickly by a healthy kidney, but more slowly or not at all by a damaged kidney.  Transplanting a damaged kidney usually results in an increase in creatinine followed by a fall back to a more normal creatinine level as the damage is healed.  When a non-damaged kidney is transplanted into an animal and there is no other kidney present, a transient peak in serum creatinine also occurs as the solitary transplanted kidney adjusts to doing the work of the two kidneys that are normally in place, but the peak is very small.  Therefore, a good way to estimate the extent of kidney damage is to look at the difference between the peak serum creatinine level seen with the damaged kidney and the transient peak that is expected with a non-damaged kidney. 

This difference is in the damage caused by preparing the kidney for cryogenic banking by replacing most of its water with chemicals called cryoprotective agents. In 2004, 21st Century Medicine was able to obtain consistent survival of transplanted rabbit kidneys that had been prepared for banking, but there was still considerable damage to these kidneys.  This was improved somewhat by 2012, but more improvement was still needed.  Last year, 21st Century Medicine was able to reduce the 2012 level of damage by nearly 50%, which was very encouraging.  But this year, even the 2015 level of damage has been cut by 83%, as of the time of this report.  This is a completely unprecedented achievement, and represents a 91% reduction of damage from the original breakthrough studies of 2004.  The amount of damage still remaining would be considered negligible by any transplant surgeon, and is far less than is seen when most human organs are transplanted today.

What this means is that it is now possible to begin to cool kidneys to deep subzero temperatures without the “overhead” of damage caused by cryoprotectant exposure.  This is expected to lead to new breakthroughs in organ banking in 2017. 

21st Century Medicine also won the Small Animal Brain Preservation Prize from the Brain Preservation Foundation in February of this year.  This represented the first time that the entire “connectome,” or “wiring pattern,” of an entire mammalian brain could be shown to be preserved for unlimited periods of time at cryogenic temperatures.  Not only was the totality of brain synaptic connections preserved, but the general structure of the brain was preserved so well that cryopreserved brains could not be visually distinguished from non-cryopreserved brains on the high-magnification electron microscope level.  In addition to providing expanded opportunities for brain mapping by neurobiologists, the new breakthrough also provides renewed hope for the potential success of cryonics.

For more information on 21CM, please keep reading below under “Cryopreservation Projects.”

Cancer Research

Every day in 2013, 1,600 Americans died of cancer,17 victims, to a great extent, of the antiquated but entrenched treatment system that relies on chemotherapy, radiation and surgery. Millions more are still alive, but survive with long-term treatment side effects, shortened life spans, and the omnipresent prospect of a cancer recurrence. Our war on cancer is just beginning.

In our quest to gain complete control over human aging, Life Extension is committed to reducing these appalling deaths from malignancies. Our support of innovative cancer research is one critical means to this end.

This cancer research progress report, authored by Orn Adalsteinsson, Ph.D., describes highlights of Life Extension’s various cancer research initiatives over the past year.

Metabolic Modulation, Immunotherapy, and Metronomic Chemotherapy

Dichloracetate , GcMAF, and Chemotherapy

Laser Assisted Immunotherapy

Based on the results of the DCA study and work looking at the combination of DCA and GcMaf, Örn Adalsteinsson, Ph.D., of the International Strategic Cancer Alliance (ISCA) and his associates, have launched a Phase I/II clinical trial to determine if the generic drug dichloracetate (DCA), combined with a low dose or metronomic chemotherapy, and a vitamin D cofactor called Gc macrophage activating factor (GcMAF) are able to induce partial &/or complete remissions in cancer subjects with a variety of malignancies. The three-month trial is taking place at a highly-regarded private clinic in the Caribbean, and is open to 20-40 participants who have failed conventional or investigational cancer therapies and have few options for further treatment. The DCA and chemotherapy are orally administered once daily for 12 weeks, while the GcMAF is injected subcutaneously once a week for 12 weeks. Participants also receive optimized nutritional supplements.

DCA is a metabolic modulator that retards the breakdown of glucose to lactic acid, interfering with the glucose uptake that is crucial to cancer cell survival. By suppressing the enzyme PDK (pyruvate dehydrogenase kinase), DCA disrupts aerobic glycolysis and essentially starves cancer cells of glucose, their primary fuel, which in turn induces apoptosis (normal cell death), decreases cancer cell proliferation, and inhibits tumor growth.18 DCA's low toxicity produces only mild side effects at effective doses, with peripheral neuropathy being the most commonly reported adverse side effect.19 Of the hundreds of studies published about DCA in the past 30 years, most relate to its use in treating the rare childhood disease congenital lactic acidosis. Only about a dozen, mostly in vitro, studies have documented DCA's efficacy as an anti-cancer agent, with several in vivo studies showing that DCA can induce apoptosis in epithelial ovarian cancer cells20 and malignant brain tumors.21 DCA was able to upregulate the apoptotic function by depolarizing the mitochondria and increasing mitochondrial reactive oxygen species.22

GcMAF has demonstrated some complete remissions on its own in patients who participated in three separate trials on breast,23 prostate,24 and colorectal cancer.25 The mechanism of action involves resupplying the Gc protein (also known as vitamin D binding protein), which cancer cells destroy by secreting an abundance of the enzyme Nagalase.26 GcMAF restores the deficiency, which is a critical component in activating the macrophages, the immune system's cancer scavengers, that in turn exert a tumoricidal action on cancer cells.27

Traditional chemotherapy regimens utilize the maximum tolerable dose with the highest acceptable toxicity (side effects) which in turn requires rest periods between cycles - a practice that not only involves re-growth of tumor cells, but also growth of selected clones resistant to the therapy. Metronomic chemotherapy (MC) is the continuous, equally spaced administration of low doses of chemotherapeutic drugs without extended rest periods. The MC treatment modality has not only been shown to be an efficacious antitumoral with very low toxicity, but also a cell target switch, aiming at tumor endothelial cells as an anti-angiogenic agent.28

While DCA , GcMAF, and MC have very different mechanisms of action - DCA restores mitochondrial metabolism which antagonizes tumor growth, metastases and survival, while GcMAF activates tumoricidal macrophages and MC acts as both an antitumoral and anti-angiogenic agent - the combination may prove to be a potent anti-cancer weapon by fighting the war simultaneously on three fronts.

Photodynamic Immunotherapy (PDIT)

For the last 8 years, Orn Adalsteinsson, Ph.D., and his research team have been conducting clinical trials exploring Laser Immunotherapy (LIT) as a possible breast cancer treatment and the results have been very promising.

In a Phase I, proof-of-concept study, 10 patients with advanced breast cancer received at least one LIT treatment. Eight patients were available for evaluation. Of those patients, one patient had a complete response (CR), four patients had a partial response (PR), two patients had progressive disease (PD) and one patient had stable disease (SD). The objective response rate was 62.5% and the clinical beneficial response rate was 75%.29

An unpublished Phase II clinical trial conducted in the Bahamas by Dr. Adalsteinsson and his research team enrolled 15 breast cancer patients who received between one and four LIT treatments. Among all 15 subjects in the study, 73.3% remain alive today. Compare that with the typical survival rate in the United States for women with advanced breast cancer, which is only 25% at 5 years.30 Currently, six subjects have surpassed the five-year milestone and of the surviving 73.3%, the average survivorship is 59.8 months or 4.9 years and counting!

Of the 15 subjects who were treated, four subjects are deceased, 11 subjects remain alive, and of those subjects four are disease free and two are in remission, which equates to a 73.3% total subject survival rate. However, of the 15 study subjects only 6 subjects completed the trial. Of those six subjects, one subject is deceased, five subjects remain alive, and of those five subjects four subjects remain disease-free, which equates to an 83.3% survival rate for subjects who completed the study.

Currently Dr. Adalsteinsson and his research team are recruiting and conducting a Phase III clinical trial examining an improved LIT technique referred to as Photodynamic Immunotherapy (PDIT). By combining a sensitizer with a corresponding laser application which is then followed by an immunoadjuvant agent and adjuvant nutritional supplements. Researchers are confident that PDIT, an enhanced LIT procedure, will greatly improve response rates in women in all stages and with varying types of breast cancer.

PDIT utilizes a photosensitizing agent which is activated by a unique wavelength of light causing one of the oxygen molecules to spin in the opposite direction in an ever-increasing arc before it returns home. This single oxygen molecule otherwise known as Singlet Oxygen or Reactive Oxygen Species (ROS) produces localized damage and moderate inflammation leading to primary tumor cell damage and the creation of tumor specific antibodies resulting in potentiation of adaptive immunity.31-33

Chemotherapy

Directly after the laser treatment, an immunoadjuvant is administered, and in the presence of localized damage and moderate inflammation created by the treatment, an immune response is initiated. The immunoadjuvant, simulated by tumor tissue fragments and cellular molecules dispersed throughout the body, act as potent inflammatory mediators rousing the body's "self-defense" system34 enabling a massive recruitment of immune cells to the damaged site.

Within minutes the mobilization of cell mediated immunity triggers a large number of neutrophils to invade the area35-36 followed by the arrival of mast cells, lymphocytes, monocytes, and macrophages with increased phagocytic capacities.35-38 Over time, after phagocytosis and tumor cell debris processing has occurred, macrophages function as antigen presenting cells39 shifting cell-mediated immunity to humoral immunity with the production of cytotoxic antibodies. These antibodies provide immunity against neoplastic cellular multiplication resulting in the destruction of locally remaining tumor cells and metastatic tumor cells as well as preventing the occurrence of new distant metastases.40-45

While the PDIT Phase III clinical trial has just begun, the research team is confident with the initial results of the work conducted thus far and are looking forward to reporting the long-term results in subsequent publications.

PET/CT Scan Reporting in Cancer Diagnosis

Enhanced PET Scan Reporting

The rapid expansion in the use of Positron Emission Tomography, or PET scans to obtain metabolic information about cancer lesions can provide oncologists and their patients with extremely valuable diagnostic and treatment management information. PET scans use an injected radioactive tracer material like fluorodeoxyglucose (FDG) to produce functional imaging that can help differentiate benign from malignant masses, evaluate tumor stage, monitor response to therapy and detect tumor recurrence in a variety of malignancies.46 Coupled with the precise anatomical imagery produced by computerized tomography, FDG PET/CT can give rapid and accurate information about tumor size, location and rate of growth.

As useful as PET imaging can be, statistical errors can at times result in “false negative” or “false positive” reporting.47 Other issues that may trigger errors include improper PET scanner calibration with patient body weight, and the variability in FDG uptake depending on the elapsed time from when the radiotracer was injected into the patient. But the most egregious errors are perhaps due to incomplete or inconsistent scan interpretations caused by inadequate training and a lack of overall standards for the quantified reporting of results. Incorrect PET scans are common today and can result in improper treatments for cancer patients.

Working with radiologist Richard Black, MD, the International Strategic Cancer Alliance adopted invaluable PET reporting practices in its Life Extension-supported laser-assisted immunotherapy breast cancer trial. Dr. Black has interpreted more than 80,000 PET/CT studies, and his methodology for an across-the-board upgrade in PET scan reporting should be incorporated at the national level to provide oncologists and their patients with the full potential PET technology has to offer. The five key features of Dr. Black’s approach will assure that oncologists receive the same kind and quality of information on each and every scan, regardless of who interpreted the scan, or where it was taken.

  1. Quantitative Reporting: Standardized uptake values, or SUV readings are collected for every object of concern in the scan, not just narrative descriptions.
  2. Reproducible Reporting: SUV readings are standardized to an area of normal homogenous tissue in the liver to generate a corrected SUV for every area of concern. The correction factor allows different experts using different equipment to obtain similar results.
  3. Index Lesion Focus: “Hotspots” indicating tumor activity must be monitored from one study to the next to enable rapid and accurate measurements of changes over time or in response to therapy.
  4. Comparative Readings Mandate: PET scan reporting must make reference to the size, SUV, and other features of an index lesion(s) from previous scans, obligating the current radiologist to request those studies for a side-by-side comparison.
  5. Image Snapshots of Index Lesions: Allows the ordering physician to visualize the areas of abnormality, rather than relying solely on a written report.

Dr. Black presented his initial findings at one of LEF’s Scientific Advisory Board Meetings in 2012; his presentation can be viewed on the Life Extension website at the following URL: www.lifeextension.com/PET-CT

Diagnostic Imaging-Combidex® - Update

Diagnostic Imaging-Combidex

The critical need to develop superior cancer imaging tools cleared a major hurdle in December 2012, when a U.S. pharmaceutical giant agreed to sell the shelved research and development rights to Combidex, a revolutionary magnetic resonance imaging (MRI) contrast agent. Combidex-enhanced scans can detect metastatic cancer lesions too small to be seen by traditional PET/CT imaging.48

Life Extension continues to be a strong advocate of Combidex since helping with the negotiation of the sale of the Combidex technology package to Radboud University Medical Center in the Netherlands in 2012. In 2013, world-renowned radiologist Jelle Barentsz, MD, with the assistance of Life Extension through Orn Adalsteinsson, has begun the process of preparing the launching of multi-country research trials, which will ultimately lead to new license applications, a commercialized product and widespread patient access.

Combidex (ferumoxtran-10) is composed of a simple sugar compound, dextran, and superparamagnetic iron oxide, or USPIO.49 These extremely small iron crystals (25-50 nanometers in diameter), become powerfully magnetized when exposed to the magnetic field of an MRI scanner. The injected Combidex contrast fluid is taken up selectively by the macrophages (scavenger cells) that are primarily found in lymph nodes and other inflammatory tissue.49, 50

Dr. Barentsz is one of the few physicians in the world to have worked extensively with Combidex technology, predominantly in prostate cancer cases. In one study, Dr. Barentsz and his team compared traditional CT scans and Combidex-enhanced MRI lymphangiography (MRL) for 375 prostate cancer patients, 16% of whom had lymph node metastases. CT imaging detected only 34% of the positive nodes, while Combidex MRL identified a remarkable 82%. The diagnoses were microscopically confirmed by either a lymph-node dissection or a needle biopsy. The study group concluded that Combidex-enhanced MRL is 96% accurate, and can eliminate the need for highly invasive surgical lymph node dissections.51

Combidex scans have also been used to successfully evaluate patients with cancers of the uterus,52 head and neck,53 kidney,54 breast,55 and liver.56

Cryopreservation Projects

Cryopreservation Projects

21st Century Medicine (21CM) is a small biotechnology company with large goals, and large accomplishments. Here are some of the recent achievements of 21st Century Medicine, none of which would have been possible without the support received from Life Extension.

Organ Banking for Transplantation

Organ transplantation could in principle save hundreds of thousands of lives every year in the United States alone, but this will require efficient means for moving organs from donor or from the laboratory to the recipient as well as better means of controlling organ rejection. Organ banking at cryogenic temperatures would solve these problems by a) enabling organs to be put “on hold” until they are needed, allowing for transportation and “just in time” use, b) allowing organs to be transplanted to ideally tissue-matched recipients rather than to poorly-matched recipients, and c) allowing time for recipients to be immunologically conditioned over several months while their specific designated organs are banked to enable the patients to receive the organs without rejection and without the need for lifelong immunosuppression. The problem has been that the technology for banking organs at cryogenic temperatures has been too difficult for most laboratories to entertain, let alone achieve. In 2013, 21CM made three breakthroughs that help to speed the day when the advantages of organ banking can be obtained by tens to hundreds of thousands of patients every year.

A New Way to Preserve Organs at Cryogenic Temperatures

21CM has pioneered a technology called “vitrification,” or glass formation, for the cryopreservation of whole organs. Vitrification enables ice formation to be avoided regardless of how much the temperature is lowered. In principle, it is the ideal way to preserve complex systems, whose cell-to-cell arrangements are damaged by the physical intrusion of ice crystals, but the problem in the case of the kidney, 21CM’s main model organ, is that the center of the kidney (the medulla), takes up the agents that prevent ice formation too slowly, which results in over-exposure of the outside of the kidney (the cortex), with resulting toxicity. But in 2013, 21CM demonstrated a new perfusion method (i.e., a method for distributing the protective agents throughout the organ using the vascular system as the means of distribution) that speeds medullary uptake of protective substances without over-exposing the cortex. Detailed comparisons of the concentrations of protective agent (cryoprotectant) that prevent ice in all parts of the kidney versus the concentrations that cause toxicity showed that, based on kidney functional testing after transplanting the kidneys, kidneys can now be made immune to ice formation without increasing toxicity. This is a major breakthrough that has taken a great many years to achieve. It is the culmination of research dating from 1980, brought to fruition only because of Life Extension's support.

A New Method for Warming Organs from Cryogenic Storage

Part of the secret of their success was defining in detail the liability of the kidney to ice formation at different warming rates. Because ice requires a certain amount of time to grow, faster warming rates result in less growth and therefore less damage. 21CM found that slow warming could not achieve sufficient ice control even using our new perfusion method, but a solution to this problem was found. Previous researchers had studied electromagnetic warming of frozen and even vitrified (though not viable) organs, but a re-analysis of the problem uncovered a superior approach that we implemented in 2013. Using the new warming technology, 21CM was able to achieve uniform warming at 160°C/min, but their whole kidney transplantation experiments showed that warming at 40-80°C/min would be sufficient for ice control after previous vitrification. 21CM believes they can achieve even faster warming in 2014, but they already have a safety margin of about 2-4 fold, which is extremely encouraging. Another advantage of the new warming method is that it turns itself off when the kidney has reached the ideal temperature for the beginning of cryoprotectant washout: the kidney heats at a maximum rate when it is most liable to form ice, and stops warming when it is free of the danger of ice formation. This technology may have many other applications, and will easily scale up to human organs.

New Technology for Preventing Fractures in Vitrified Organs

Despite the beauty of vitrification as a method of cryopreservation of whole organs, it introduces a danger of crack formation below the temperature at which the organs revert from the liquid state to the glassy state (the glass transition temperature, at which biological change is essentially arrested as a result of the lack of mobility of the molecules in the living system). At the glass transition temperature, due to the lack of motion of the molecules in the system, thermal contraction stress cannot be relieved, so further cooling builds up stress and may result in cracking of the organ. This problem had been poorly studied in the past, but is essential to ensure the safety of cryogenic organ banking. 21CM found experimentally in 2013 that every organ in the body can be cooled to 6°C below the glass transition temperature without forming any cracks. 21CM even found that we could cool a liter of cryoprotectant solution to the temperature of liquid nitrogen without fracturing, and this liter of solution is expected to be more liable to crack than would be a whole organ, and is about as voluminous as most human organs.

Together, these observations poise 21CM for successful demonstration of kidney banking in 2014. 21CM also developed a pig model at 21CM that will allow them to vitrify and transplant not just rabbit kidneys (their model in 2013), but also pig kidneys, which are as large as human kidneys and should pave the way for application of 21CM technology in human clinical medicine.

Cornea Banking for Transplantation

Several years ago, 21CM demonstrated successful banking of human corneas, as shown by vital staining, light and electron microscopy, and transplantation into primates. What was missing was a feasible way of translating this accomplishment to the clinic. In 2013, 21CM was approached by a major eye institute about the possibility of performing human clinical trials. 21CM was requested to re-demonstrate their method using another measure of success prior to beginning human transplantation. 21CM chose the ability of the cornea to maintain its hydration during in vitro superfusion for up to about 30 hours, which is a common “acid test” of corneal function in vitro. Vitrified corneas performed nearly as well in this assay system as control corneas obtained from a cooperating eye, opening the door to human transplants in 2014. The benefits of the ability to bring sight to the blind all around the world, which is not presently possible given deterioration of control corneas during transportation outside the US, are self-explanatory. There is no other laboratory, or any other technology, that has been able to reproducibly preserve human corneas after vitrification, and the only theoretically competing method is not practical and is not being commercially pursued. Freezing is no longer used as a method of corneal banking due to its poor long-term effects.

Keeping the Brain Alive in the Cold

Alzheimer’s Research

In the last century, major advances in clinical hypothermia enabled previously intractable surgical problems, such as the ability to correct cerebral aneurysms, to be addressed for the first time. Still, the procedure has had its hazards, and, apart from one much older and non-definitive paper, no published method exists that allows the brain and the rest of the body to be put “on hold” for more than 3 hours, which may be inadequate for many purposes. In 2013, 21CM made major breakthroughs on hypothermic brain preservation, on the theory that the brain is both the weakest link in the whole body chain and the least studied organ in the body in terms of the effects of prolonged hypothermia. Through an extensive series of optimizations, and the use of novel pharmacological agents for this purpose, 21CM was able to preserve whole rabbit brains for 15 hours by continuous hypothermic perfusion with complete recovery of electrical in all brain regions, and with no diminution in perfusion rate over 15 hours. Previous investigation of brain ultrastructure showed excellent results even before the current advances, so 21CM believes ultrastructure is well preserved as well. Preliminary results after even 24 hours of preservation have been very encouraging as well, and even equal to non-preserved control brain results. 21CM began construction of equipment to enable testing of whole brain viability after hypothermic preservation in 2013 and completed and implemented this equipment in 2014.

21CM believes these results could enable the rescue of trauma victims and soldiers who cannot be helped with presently available technology. To further explore this possibility, 21CM is establishing a method for 24-hour hypothermic preservation of whole 80-kg pigs, and experiments are imminent. 21CM’s initial results will focus on perfusion rates, edema, histological integrity, and ultrastructural preservation, but 21CM will seek additional funding from an outside funding agency for more detailed studies of energy metabolism and the reversibility of extended hypothermic perfusion in whole large mammals. 21CM’s principle is that if they can preserve whole animals or brain preparations for very prolonged periods, there will be greater comfort in applying these methods under more critical circumstances for lesser periods of time given the very large margin of safety of the technology.

References

  1. Buffenstein R. The naked mole-rat: a new long-living model for human aging research. J Gerontol A Biol Sci Med Sci. 2005 Nov;60(11):1369-77.
  2. Azpurua J, Seluanov A. Long-lived cancer-resistant rodents as new model species for cancer research. Front Genet. 2013 Jan 9;3:319.
  3. Keane M, Craig T, Alföldi J, Berlin AM, Johnson J, Seluanov A, Gorbunova V, Di Palma F, Lindblad-Toh K, Church GM, de Magalhães JP. The Naked Mole Rat Genome Resource: facilitating analyses of cancer and longevity-related adaptations. Bioinformatics. 2014 Dec 15;30(24):3558-60.
  4. Tian X, Azpurua J, Hine C, Vaidya A, Myakishev-Rempel M, Ablaeva J, Mao Z, Nevo E, Gorbunova V, Seluanov A. High-molecular-mass hyaluronan mediates the cancer resistance of the naked mole rat. Nature. 2013 Jul 18;499(7458):346-9.
  5. Azpurua J, Ke Z, Chen IX, Zhang Q, Ermolenko DN, Zhang ZD, Gorbunova V, Seluanov A. Naked mole-rat has increased translational fidelity compared with the mouse, as well as a unique 28S ribosomal RNA cleavage. Proc Natl Acad Sci U S A. 2013 Oct 22;110(43):17350-5.
  6. Breakthrough of the year 2013. Notable developments. Science. 2013 Dec 20;342(6165):1435-41.
  7. Gorbunova V, Seluanov A, Zhang Z, Gladyshev VN, Vijg J. Comparative genetics of longevity and cancer: insights from long-lived rodents. Nat Rev Genet. 2014 Aug;15(8):531-40.
  8. MacRae SL, Zhang Q, Lemetre C, Seim I, Calder RB, Hoeijmakers J, Suh Y, Gladyshev VN, Seluanov A, Gorbunova V, Vijg J, Zhang ZD. Comparative analysis of genome maintenance genes in naked mole rat, mouse, and human. Aging Cell. 2015 Apr;14(2):288-91.
  9. Mao Z, Hine C, Tian X, Van Meter M, Au M, Vaidya A, Seluanov A, Gorbunova V. SIRT6 promotes DNA repair under stress by activating PARP1. Science. 2011 Jun 17;332(6036):1443-6.
  10. Kines KJ, Sokolowski M, deHaro DL, Christian CM, Belancio VP. Potential for genomic instability associated with retrotranspositionally-incompetent L1 loci. Nucleic Acids Res. 2014;42(16):10488-502.
  11. Belancio VP, Blask DE, Deininger P, Hill SM, Jazwinski SM. The aging clock and circadian control of metabolism and genome stability. Front Genet. 2015 Jan 14;5:455.
  12. Blask DE, Dauchy RT, Dauchy EM, Mao L, Hill SM, Greene MW, Belancio VP, Sauer LA, Davidson L. Light exposure at night disrupts host/cancer circadian regulatory dynamics: impact on the Warburg effect, lipid signaling and tumor growth prevention. PLoS One. 2014 Aug 6;9(8):e102776.
  13. Xiang S, Dauchy RT, Hauch A, Mao L, Yuan L, Wren MA, Belancio VP, Mondal D, Frasch T, Blask DE, Hill SM. Doxorubicin resistance in breast cancer is driven by light at night-induced disruption of the circadian melatonin signal. J Pineal Res. 2015 Aug;59(1):60-9.
  14. deHaro D, Kines KJ, Sokolowski M, Dauchy RT, Streva VA, Hill SM, Hanifin JP, Brainard GC, Blask DE, Belancio VP. Regulation of L1 expression and retrotransposition by melatonin and its receptor: implications for cancer risk associated with light exposure at night. Nucleic Acids Res. 2014 Jul;42(12):7694-707.
  15. Hill SM, Belancio VP, Dauchy RT, Xiang S, Brimer S, Mao L, Hauch A, Lundberg PW, Summers W, Yuan L, Frasch T, Blask DE. Melatonin: an inhibitor of breast cancer. Endocr Relat Cancer. 2015 Jun;22(3):R183-204.
  16. Sokolowski M, DeFreece CB, Servant G, Kines KJ, deHaro DL, Belancio VP. Development of a monoclonal antibody specific to the endonuclease domain of the human LINE-1 ORF2 protein. Mob DNA. 2014 Dec 10;5(1):29.
  17. Available at: http://www.cancer.org/research/cancerfactsfigures/cancerfactsfigures/cancer-facts-figures-2013 Accessed February 19, 2014
  18. Bonnet S, Archer SL, Allalunis-Turner J, et al. A mitochondria-K+ channel is suppressed in cancer and its normalization promotes apoptosis and inhibits cancer growth. Cancer Cell. 2007 Jan;11(1):37-51.
  19. Felitsyn N, Stacpoole PW, Notterpek L. Dichloroacetate causes reversible demyelination in vitro: potential mechanism for its neuropathic effect. J Neurochem 100:429-436, 2007.
  20. Wong JY, Huggins GS, Debidda M, et al. Dichloracetate induces apoptosis in endometrial cancer cells. Gynecol Oncol. 2008;109:394-402.
  21. Michelakis ED, Sutendra G, Dromparis P, et al. Metabolic modulation of glioblastoma with dichloracetate. Sci Transl Med. 2010 May 12;2(31)ra34.
  22. Strum SB, Adalsteinsson O, Black RR, et al. Case Report: Sodium Dichloracetate (DCA) inhibition of the "Warburg Effect" in a human cancer patient: complete response in non-Hodgkin's lymphoma after disease progression with rituximab-CHOP. J Bioenerg Biomembr. 2012 Dec 20 (Epub ahead of print).
  23. Yamamoto N, Suyama H, Yamamoto N, Ushijima N. Immunotherapy of metastatic breast cancer patients with vitamin D-binding protein-derived macrophage activating factor (GcMAF). Int J Cancer. 2008 Jan 15;122(2):461-7.
  24. Yamamoto N, Suyama H, Yamamoto N. Immunotherapy for Prostate Cancer with Gc Protein-Derived Macrophage-Activating Factor, GcMAF. Trans Oncol. 2008 Jul;1(2):65-72.
  25. Yamamoto N, Suyama H, Nakazato H, Yamamoto N, Koga Y. Immunotherapy of metastatic colorectal cancer with vitamin D-binding-protein-derivedd macrophage-activating factor, GcMAF. Cancer Immunol Immunother. 2008 Jul;57(7):1007-16.
  26. Mohamad SB, Nagasawa H, Uto Y, Hori H. Tumor cell alpha-N-acetylgalactosaminidase activity and its involvement in GcMAF-related macrophage activation. Comp Biochem Physiol A Mol Integr Physiol. 2002 May;132(1):1-8.
  27. Thyer L, Ward E, Smith R. A novel role for a major component of the vitamin D axis: vitamin D binding protein-derived macrophage activating factor induces human breast cancer cell apoptosis through stimulation of macrophages. Nutrients. 2013 Aug;5(7):2577-89.
  28. Scharovsky OG, Mainette LE, Razados VR. Metronomic chemotherapy: changing the paradigm that more is better. Curr Oncol. 2009 Mar;16(2):7-15.
  29. Li X, Ferrel GL, Guerra MC, et al: Preliminary safety and efficacy results of laser immunotherapy for the treatment of metastatic breast cancer patients. Photochem. Photobiol. Sci. 2011, 10: 817-821.
  30. Available at: http://www.cancer.org/research/cancerfactsfigures/cancerfactsfigures/cancer-facts-figures-2012 Accessed January 12, 2013.
  31. Wilson BC: Photodynamic therapy for cancer: principles. Canadian Journal of Gastroenterology. 2002, 16(6): 393-396.
  32. Dolmans DE, Fukumura D, Jain RK: Photodynamic therapy for cancer. Nature Reviews Cancer. 2003, 3(5):380-387.
  33. van Duijnhoven FH, Aalbers RI, Rovers JP, et al: The immunological consequences of photodynamic treatment of cancer, a literature review. Immunobiol. 2003,207, 105 - 113.
  34. de Vree WJ, Essers MC, de Bruijn HS, et al: Evidence for an important role of neutrophils in the efficacy of photodynamic therapy in vivo. Cancer Res. 1996,56(13):2908-11.
  35. Krosl G, Korbelik M and Dougherty GJ: Induction of immune cell infiltration into murine SCCVII tumour by photofrin-based photodynamic therapy. Br. J. Cancer. 1995, 71: 549-555.
  36. Gollnick SO, Liu X, Owczarczak B, et al: Altered expression of interleukin 6 and interleukin 10 as a result of photodynamic therapy in vivo. Cancer Res. 1997, 57: 3904-3909.
  37. Yamamoto N, Sery TW, Hoober JK, et al: Effectiveness of photofrin II in activation of macrophages and in vitro killing of retinoblastoma cells. Photochem. Photobiol. 1994, 60: 160-164.
  38. Ziegler K and Unanue ER: Identification of a macrophage antigen-processing event required for Iregion-restricted antigen presentation to T lymphocytes. J. Immunol. 1981, 127: 1869-1875.
  39. Coutier S, Bezdetnaya L, Marchal S, et al: (mTHPC) photosensitized macrophage activation: enhancement of phagocytosis, nitric oxide release and tumour necrosis factor-alpha-mediated cytolytic activity. Br. J. Cancer. 1999, 81: 37- 42.
  40. Chen W R, Zhu, W-G, Dynlacht, J R, et al: Long-term tumor resistance induced by laser photo-immunotherapy. Int. J. Cancer. 1999, 81: 808-812.
  41. Li X and Chen WR: Laser immunotherapy: novel modality to treat cancer through specific antitumor immune response. Zhongguo Jiguang/Chinese Journal of Lasers. 2010, 37 (11): 2698-2702.
  42. Li X, Naylor MF, Le H, et al: Clinical effects of in situ photoimmunotherapy on late-stage melanoma patients. Cancer Bio & Ther. 2010a, 10 (11):, 1081-1087.
  43. Peniche H and Peniche C: Chitosan nanoparticles: a contribution to nanomedicine. Polymer Internat. 2011, 60(6):883-889.
  44. Li X, Gu Y, Du N, et al: Laser immunotherapy: Concept, possible mechanism, clinical applications, and recent experimental results. IEEE J. Sel. Top. Quantum Electron. 2012, 18: 1434-1438.
  45. Ferrel GL, Zhou F, Li X, et al: Effects of laser immunotherapy on late-stage, metastatic breast cancer patients in a Phase II clinical trial. Biophotonics and Immune Responses. 2014, 8944: 89440I.
  46. Yoon KT, Kim JK, Kim do Y, et al. Role of 18F-flourodeoxyglucose positron emission tomography in detecting extrahepatic metatastasis in pretreatment staging of hepatocellular carcinoma. Oncology. 2007;72 Suppl 1:104-10.
  47. Black RR. Optimization of FDG PET-CT imaging in oncology 2012 (Power point presentation).
  48. Harisinghani MG, Barentsz J, Hahn P, et al. Noninvasive Detection of Clinically Occult Lymph-Node Metastases in Prostate Cancer. N Engl J Med. 2003 Jun;348:2491-2499.
  49. Barentsz JO, Futterer JJ, Takahashi S. Use of ultrasmall superparamagnetic iron oxide in lymph node MR imaging in prostate cancer patients. Eur J Radiol. 2007 Sep;63(3):369-72.
  50. Corot C, Robert P, Idée JM, Port M. Recent advances in iron oxide nanocrystal technology for medical imaging. Adv Drug Deliv Rev. 2006 Dec 1;58(14):1471-504.
  51. Heesakkers RA, Hovels AM, et al. MRI with a lymph-node-specific contrast agent as an alternatie to CT scan and lymph-node dissection in patients with prostate cancer: a prospective multicohort study. Lancet Oncol. 2001 Sep;9(9):850-6.
  52. Laghi A, Paolantonio P, Panebianco V, et al. Decrease of signal intensity of myometrium and cervical stroma after ultrasmall superparamagnetic iron oxide (USPIO) particles administration: an MR finding with potential benefits in T staging of uterine neoplasms. Invest Radiol. 2004 Nov;39(11):666-70.
  53. Curvo-Semedo L, Diniz M, Miguéis J, et al. USPIO-enhanced magnetic resonance imaging for nodal staging in patients with head and neck cancer. J Magn Reson Imaging. 2006 Jul;24(1):123-31.
  54. Guimaraes AR, Tabatabei S, Dahl D, et al. Pilot study evaluating use of lymphotrophic nanoparticle-enhanced magnetic resonance imaging for assessing lymph nodes in renal cell cancer. Urology. 2008 Apr;71(4):708-12.
  55. Daldrup-Link HE, Rydland J, Helbich TH, et al. Quantification of breast tumor microvascular permeability with feruglose-enhanced MR imaging: initial phase II multicenter trial. Radiology. 2003 Dec;229(3):885-92.
  56. Yoo HJ, Lee JM, Lee MW, et al. Hepatocellular carcinoma in cirrhotic liver: double-contrast-enhanced, high-resolution 3.0T- MR imaging with pathologic correlation. Invest Radiol. 2008 Jul;43(7):538-46.