By Alan Cocchetto, NCF Medical Director ©2011

From Summer 2011 Forum

It has been over two years since the NCF approached scientists, at Cairo University as well as those at the Weizmann Institute of Science, offering them unique research opportunities to study CFIDS/ME disease endpoints. This article provides patients with a progress report regarding the scientific research accomplished by both groups.

Background:

Patient bone-marrow biopsies, pathology and flow-cytometry studies have indicated that one of the end points of CFIDS/ME is myelodysplasia. Myelodysplasia, also known as Myelodysplastic Syndrome (MDS), is a pre-leukemia condition that involves the ineffective production of the myeloid class of blood cells. Patients can develop cytopenias (low blood counts) and anemia. A percentage of these patients will go on to develop AML or acute myelogenous leukemia ^[1]. One key factor that influences this disease process is through the generation of elastase which has been measured in CFIDS/ME patients. Elastase has been found to be highly abnormal in these patients ^[2-4].

Foreground: Cairo University Studies

The lead researcher, Dr. Hany El-Shemy, has a double PhD in biochemistry and genetic engineering and is full professor at Cairo University. His NCF's research proposal was titled "Production of natural biological active formulas as anti-carcinogenic AML drugs."

Dr. El-Shemy's scientific team focused on evaluating various bioactive compounds as well as numerous natural products to study their effectiveness, in-vitro, against several AML leukemia cell lines. Adding to the uniqueness of this research was the fact that his team looked at and subsequently scored the effectiveness of these compounds against current commercially available drug treatments. This research was extended to subsequently evaluate compound synergy. In other words, several compounds would first be combined and then evaluated in-vitro against their drug counterparts. A list was then created that further highlighted their additive effects.

Some additional background: Cairo University Studies

We have been asked why the NCF would fund leukemia studies? Actually, the answer is very simple. Since the NCF had direct evidence for myelodysplasia development in CFIDS/ME patients who also had very high levels of elastase, an immediate connection was made. The NCF had patients who didn't qualify for chemotherapy for MDS/AML due to the fact that their pathology samples hadn't exceeded certain disease thresholds established by the oncology world committees. Simply stated, CFIDS/ME patients were considered to have what is properly referred to as "minimal residual disease" in spite of the fact that cancerous cells were identified in their bone-marrow samples. Think about this for a minute if you will. Your oncologist tells you that they can't treat the cancerous cells because you don't have enough of them! In other words, you haven't exceeded the threshold required for treatment. So if you have say 8% abnormal cells but the threshold is set at 15% abnormal cells required for treatment, then you are placed in a wait-and-see mode. The NCF considers this most unfortunate to say the least. This is especially difficult because of the nature of myelodysplasia and its smoldering disease process. As a direct result, the NCF went on a mission to help patients who were at this point in their disease process; an unfortunate one that we have seen time and time again. What resulted was some truly innovative research that was completed by Dr. El-Shemy and his team of very dedicated scientists at Cairo University.

Cairo University — Nuts and bolts:

The NCF had personally identified and selected ten compounds which would be formally evaluated in this research study. This selection was based on previous CFIDS/ME patient studies that had been undertaken by the NCF. Dr. El-Shemy's team subsequently added several compounds to this list for testing as well.

To reiterate, these compounds were evaluated both individually and combinationally, in-vitro, in order to define their mechanisms of action regarding their anti-leukemic effect. The most promising results were then analyzed using chemo-informatics analysis by experts in this field. Target profiling using the Ontomine, Docking and Gene Expression Network analysis indicated that these compounds target proteins which are critically important for cancer treatment. For this NCF research, the National Cancer Institute supplied 182 bone-marrow patient (AML) samples. In addition, a parallel analysis of benchmarked drugs, which exist in the market for AML treatments, was completed so that the predictions for effectiveness could be supported or refuted by published work. Furthermore, analysis on benchmark drugs was successful in predicting the effectiveness of well known drug combinations used for AML treatments.

Thus far, four medical journal publications have resulted directly from this research while three others have been submitted for publication. Some of these are listed in the reference section of this article ^[5-7].
In addition, Dr. El-Shemy was a presenter at an international conference. His lecture reflected the NCF's efforts in this research area ^[8].

More foreground: Weizmann Institute of Science Studies

The NCF approached Dr. Tsvee Lapidot at the Weizmann Institute of Science, in Israel, to see if he had an interest in pursuing research to test various elastase inhibitor compounds to analyze their effects on stem cells and AML. Dr. Lapidot, who is a professor of immunology and chair of stem cell research at the Weizmann Institute, has an extensive background in the role of elastase in hematopoiesis and AML development. To this end, Dr. Lapidot's research, funded by the NCF, was titled "Involvement of elastase and SDF-1/CXCR4 axis in regulation of motility and development of human AML initiating cells."

More background: Weizmann Institute of Science Studies

Much like the Cairo University studies, the NCF had searched for elastase inhibitors to be evaluated by Dr. Lapidot and his team of scientists. Dr. Lapidot studied the capability of ten selected compounds to affect key functional activities of AML cells in-vitro. These tests included various assays that studied cell motility, adhesion to bone-marrow stromal cells and proliferation.

One of the interesting notes, regarding AML cells, is that they express a chemokine receptor known formally as CXCR4. This expression is crucial for their migration, adhesion, proliferation, homing, engraftment and subsequent egress to the circulation. Moreover, high CXCR4 expression level in leukemic cells has been proven to be a predictor of overall survival and relapse-free survival in patients with AML thus underlying a profound involvement of CXCR4 in the regulation of AML. CXCR4 works in conjunction with another factor, stromal cell-derived factor-1 (SDF-1), whose axis is critical to myeloid leukemia development.

Why might all of this be important to CFIDS/ME patients? There have been numerous reports of an association between active HHV-6 infections and CFIDS/ME. Probably the most noted one was published by Buchwald, the lead author ^[9]. As it turns out, marked downregulation of CXCR4 has been reported by HHV-6A, HHV-6B and HHV-7 viruses ^[10]. Thus, these viruses are very capable of directly impacting the CXCR4/SDF-1 axis. A concurrent infection with any of these viruses would thereby be expected to have an effect on leukemic cell survival and therefore, patient survival.

Since there are additional on-going efforts with Dr. Lapidot's group at this time, the NCF cannot comment on any further developments.

References:

1. Myelodysplastic syndrome; http://en.wikipedia.org/wiki/Myelodysplastic_syndrome
2. Unravelling intracellular immune dysfunctions in chronic fatigue syndrome: interactions between protein kinase R activity, RNase L cleavage and elastase activity, and their clinical relevance; Meeus M, Nijs J, McGregor N, Meeusen R, De Schutter G, Truijen S, Frémont M, Van Hoof E, De Meirleir K; In Vivo. 2008 Jan-Feb;22(1):115-21
3. Diagnostic methods for measuring elastase activity levels; Inventors: Englebienne P, DeMeirleir K, Herst CV; US Patent Application # 20080193957, August 14, 2008
4. Methods of treatment of chronic immune disease; Inventors: El Bakkouri K, Englebienne P, DeMeirleir K, Herst CV; US Patent Application # 20050032770, February 10, 2005
5. Antileukemia activity from root cultures of Vernonia amygdalina; Mutasim M. Khalafalla, Eltayb Abdellatef, Hussein M. Daffalla, Amr A. Nassrallah, Khalid M. Aboul-Enein, David A. Lightfoot, Alan Cocchetto and Hany A. El-Shemy; Journal of Medicinal Plant Research 3, 556-562. 2009
6. Anti-cancer and anti-oxidant activity of some Egyptian medicinal plants; Amr A. Nassr-Allah, Ahmed M. Aboul-Enein, Khalid M. Aboul-Enein, David A. Lightfoot, Alan Cocchetto and Hany A. El-Shemy; Journal of Medicinal Plant Research 3(10): 799-808. 2009
7. Antitumor Properties and Modulation of Antioxidant Enzymes’ Activity by Aloe vera Leaf Active Principles Isolated by Supercritical Carbon Dioxide Extraction; H.A. El-Shemy, M.A.M. Aboul-Soud, A.A. Nassr-Allah, K.M. Aboul-Enein, A. Kabash and A. Yagi; Current Medicinal Chemistry, 17. 129-138. 2010
8. Hany A. El-Shemy; Lecture was given at the Annual World Congress on "Bioequivalence & Bioavailability-Pharmaceutical R&D Summit (BIOBIO-2010)", March 1-3, 2010 in Hyderabad, India, a speech on “In Silico Selection of Natural Anti-leukaemia Compounds” in Track 2-3 “Biosimulation in BE Studies-Prediction Tools.”
9. A chronic illness characterized by fatigue, neurologic and immunologic disorders, and active human herpesvirus type 6 infection; Buchwald D, Cheney PR, Peterson DL, Henry B, Wormsley SB, Geiger A, Ablashi DV, Salahuddin SZ, Saxinger C, Biddle R, et al.; Ann Intern Med. 1992 Jan 15;116(2):103-13.
10. Down-regulation of CXCR4 by human herpesvirus 6 (HHV-6) and HHV-7; Yasukawa M, Hasegawa A, Sakai I, Ohminami H, Arai J, Kaneko S, Yakushijin Y, Maeyama K, Nakashima H, Arakaki R, Fujita S; J Immunol. 1999 May 1;162(9):5417-22.