The Critical Mission of Life Extension Foundation®
Leukocyte Infusion Therapy
Leukocyte infusion therapy is one of the new cancer immunotherapies designed to stimulate the immune system to destroy tumors. The original animal studies were carried out by Dr. Zheng Cui of Wake Forest University, who found that a subpopulation of leukocytes (white cells) called granulocytes are primarily responsible for killing cancer cells. In order to evaluate leukocyte infusion therapy in humans, Life Extension Foundation provided two large grants for Phase I/II clinical trials. These trials are now under way for a maximum of 29 patients with metastatic, nonhematological cancers under the direction of Dipnarine Maharaj, M.D., at the South Florida Bone Marrow/Stem Cell Transplant Institute in Boynton Beach, Florida.
Granulocytes from the blood of healthy human donors are concentrated for infusion into the study participants. Because four to six donors are needed for each subject, a Donor Registry was established with strict eligibility criteria, including screening for infectious diseases and other blood testing; approximately 100 donors are currently registered.
To date, eight cancer patients with advanced solid tumors of the colon, breast, esophagus, ovaries and prostate have undergone treatment of which two have completed the treatment. Each subject received a total of four granulocyte transfusions, which were well-tolerated. The protocol calls for the treatment of 20 study subjects before any conclusions can be drawn about the effectiveness of the therapy. The trial’s four endpoints are dose response at 90 days, tolerance, safety and efficacy. The response at 90 days after the last infusion will be based on comparison of tumor measurements at baseline. Given the success of the original animal studies conducted at Wake Forest University which were able to cure animals of cancer, Dr. Maharaj is seeking similar results when leukocyte infusion therapy is used in humans.
Currently, the Life Extension Foundation is sponsoring a phase II clinical trial investigating DCA, short for dichloroacetic acid, for individuals who have stage IV cancer with solid tumors or prostate cancer and have failed conventional or investigational therapies, with limited options for further therapy.
DCA is a metabolic modulator which has been used for the treatment of lactic acidosis and inherited diseases of mitochondrial metabolism in humans for over 30 years. Preclinical in vitro and in vivo models have demonstrated the DCA's beneficial potential in human cancers and there is 40 years of experience with mechanistic studies looking at human tissues after oral consumption, pharmacokinetic, and toxicity data.
Early Stages of the DCA Clinical Trial
There is also data from 6-month randomized studies and 5-year use case reports, all which has supported early-phase clinical trials. Further, prior case studies and clinical trials have reported response to DCA in leukemia and lymphoma, prostate, breast, lung, and brain and nervous system cancers. The metabolic profile of malignancy, or unopposed cell growth is one associated with various metabolic adaptations that preferentially utilize pathways involved with glycolysis, called the glycolytic phenotype (GP) of cancer. In this adaptation, the cancer cell diminishes, as well as undermines, the metabolic pathways of glucose oxidation (GO) used by normal cells for energy production as well as tumor cell elimination. One such function that is compromised involves the important mitochondrial function of programmed cell death (apoptosis). DCA, in effect, throws the metabolic switch so that cellular metabolism is side-tracked from the GP, and back to GO. In doing so, DCA restores the mitochondria's apoptotic function and the ability to impair tumor cell proliferation.
To date, the DCA trial has recruited less than ¼ (or 6/40 subjects) of its evaluable 40 subjects mandatory to fulfill the study requirements. While there has been some evidence of tumor response to DCA there has also been some disease progression as well. However, it is early in the study process and there is insufficient data to evaluate or draw any conclusions thus far.
PET/CT Scan Reporting in Cancer Diagnosis
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.
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.19 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 atone of LEF's Scientific Advisory Board Meetings in 2012; his presentation can be viewed on the Life Extension website at the following URL: www.LEF.org/PET-CT
Diagnostic Imaging-Combidex® - Update
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.
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 Foundation 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. 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.
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.
Combidex scans have also been used to successfully evaluate patients with cancers of the uterus, head and neck, kidney, breast, and liver.
Data published in 2011 also showed the benefit of perispinal Enbrel® in restoring cognitive function in three consecutive patients with stable and persistent chronic neurological deficits due to strokes that had failed to resolve despite previous treatment and rehabilitation. The onset of clinical response was evident within 10 minutes of perispinal injection in all patients, including improvements in hemiparesis, gait, hand function, hemi-sensory deficits, spatial perception, speech, cognition and behavior.
Enbrel® is an expensive drug and administering it in a way that delivers it to the brain is an intricate medical process. The Life Extension Foundation has been sponsoring a study being conducted in Florida for individuals diagnosed with mild to moderate Alzheimer’s disease. The objective of the research initiative is to measure the effects of weekly Enbrel® injections plus nutritional supplements that may suppress the inflammatory factor implicated in the neuronal degeneration of Alzheimer’s disease.
As of August 2014, only one more patient is needed to complete this study. To participate in this study register now online at: www.lef.org/clinicalresearch or send an e-mail expressing your interest in participating in this study to:LEClinicalResearch@LifeExtension.com. For any further information you may call the Life Extension Clinical Research Department at (866) 517-4536 between 9:00 a.m. and 5:00 p.m. ET or leave a voicemail after hours with your name and contact information.
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 the Life Extension Foundation.
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 Foundation 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
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.
More than 5 million Americans are stricken with Alzheimer’s disease today. One out of three Americans over age 85 is diagnosed with Alzheimer’s. As the aging population increases, the prevalence will approach 13 to 16 million cases in the United States by 2050.
A startling discovery revealed that an FDA-approved autoimmune disease drug (Enbrel®) may reverse the clinical course of Alzheimer’s disease by blocking a pro-inflammatory cytokine tumor necrosis factor-alpha (TNF-a) that damages the brain. Alzheimer’s patients who had this drug perispinal extrathecal-administered in the posterior neck, delivering Enbrel® to the brain via the cerebrospinal venous system, experienced remarkable cognitive improvements in minutes.
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 will complete and implement 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.
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.
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