Protein Discovered That Keeps Hemoglobin In Balance Research May Advance Treatme

Protein Discovered That Keeps Hemoglobin In Balance; Research May Advance Treatment Of The Blood Disease Thalassemia
Source: Children's Hospital of Philadelphia
June 13, 2002

Hematology researchers at The Children's Hospital of Philadelphia have discovered a gene and its associated protein that may have major implications for red blood cell formation, specifically for hemoglobin, which carries oxygen in red blood cells. Understanding how this protein functions may eventually lead to novel treatments for the hemoglobin-related blood disease, thalassemia.

Thalassemia is a group of related inherited disorders that together are the most common single-gene disease known. The most severe form of the disease, beta thalassemia major, affects 300,000 patients worldwide.

Thalassemia results from an imbalance between two proteins in hemoglobin, called alpha and beta globin. An excess of either type of protein is toxic, causing thalassemia symptoms including poor growth, fatigue, bone damage, or skin ulcers. The newly found protein, alpha hemoglobin stabilizing protein (AHSP), binds to free alpha globin and prevents it from forming a precipitate that damages red blood cells.

"AHSP acts as a chaperone molecule - a chemical that helps another protein to fold or unfold," said Mitchell Weiss, M.D., Ph.D., a pediatric hematologist at The Children's Hospital of Philadelphia, and senior author of the paper, published in the June 13 issue of Nature. "Here it makes free alpha globin stable and prevents its deleterious effects."

Dr. Weiss' team suspected that AHSP, by preventing free alpha hemoglobin from precipitating within red blood cells, protects the cells from injury. To test that hypothesis, the researchers developed knockout mice, animals genetically engineered to lack the gene that produces AHSP. Those mice showed blood abnormalities similar to those found in mice with thalassemia.

AHSP's protective role could explain how some patients who carry the genetic trait for beta thalassemia have mild disease and few symptoms even though their bodies produce more alpha than beta globin. By binding to free alpha globin, AHSP may protect the body from a dangerous accumulation of that protein. However, if AHSP does not function properly, the excess alpha globin precipitation may change milder or intermediate thalassemia into more severe disease.

This current research suggests that if physicians can deliver AHSP or a similar agent to patients with thalassemia, they may produce a new treatment for the disease. Severe cases are now treated with frequent blood transfusions that carry their own serious complications, such as excess iron. "If we can reduce the buildup of free alpha globin we may be able to lower the dose of transfusion needed, and improve patients' quality of life," added Dr. Weiss.

In addition to Dr. Weiss, co-authors of the article are Anthony J. Kihm, Ph.D., Yi Kong, Wei Hong, Ph.D., J. Eric Russell, M.D., Susan Rouda, Kazuhiko Adachi, Ph.D., and Gerd A. Blobel, M.D., Ph. D., all of Children's Hospital's Division of Hematology, and M. Celeste Simon, Ph.D., of the Howard Hughes Medical Institute and the University of Pennsylvania. The research was funded by the Cooley's Anemia Foundation, the Unico Foundation and the National Institutes of Health.

Founded in 1855 as the nation's first pediatric hospital, The Children's Hospital of Philadelphia is ranked today as the best pediatric hospital in the nation by a comprehensive Child magazine survey. Its pediatric research program is among the largest in the country, ranking second in National Institutes of Health funding.

Excessive α-Hb is particularly damaging, as evidenced by β-thalassemia, a common inherited anemia in which mutations in the β-globin gene impair the production of β-Hb with consequent buildup of the unpaired α-subunit (1–5). Intact monomeric α-Hb generates ROS [Reactive oxygen species (ROS) include oxygen ions, free radicals, and peroxides] that damage cellular proteins, lipids, and nucleic acids (6). In addition, α-Hb is structurally unstable, with a tendency to denature upon oxidation, filling the cytoplasm and cell membrane with precipitated α-globin polypeptides, free heme, porphyrins, and iron, which further propagate ROS production (reviewed in ref. 7). Together, these effects reduce the lifespan of circulating erythrocytes and also impair the viability of erythroid precursors in hematopoietic tissues, causing ineffective erythropoiesis.

Clinical efficacy and molecular mechanism of nourishing shen and supplementing marrow principle in treating β-thalassemia
Wu Zhi-kui1 Contact Information, Fang Su-ping1, Zhang Xin-hua1, Cai Hui-guo3, Wang Lei1, Yi Jie1, Chai Li-min1, Lu Xin-xia1, Chen Yu-ying1, Huang You-wen2, Wang Rong-xin2 and Chen Pei-zhen3
(1)     Department of Molecular-Biology, Guang’anmen Hospital, China Academy of Traditional Chinese Medicine, 100053 Beijing
(2)     The 303 Hospital of PLA, China
(3)     Institute of Hematology, Chinese Academy of Medical Sciences, China

Received: 8 June 2003 
Abstract   Objective: To explore the possibility of using traditional Chinese medicine (TCM) in treating β-thalassemia, its clinical effect and molecular mechanism of the action.Methods: According to the TCM theory of “Shen producing marrow”, the composite recipe, Yisui Shenxueling Granule (YSSXL), consisting of Chinese drugs for nourishing Shen and supplementing marrow (NS & SM) was given orally to 78 patients with β-thalassemia (49 of the severe type and 29 of moderate type), 3 times a day, 10 g each time (for children, the dose would be reduced properly), with 3 months as one therapeutic course, and no blood transfusion used in the course. The clinical therapeutic efficacy and hematologic parameters in patients were observed, and systemic gene analysis was conducted with PAGE, PCR, PCR-SSCP, RT-PCR and DNA sequences analysis and mRNA detection, in order to study the molecular mechanism from the relationships between genetic mutation and clinical efficacy, gene expression and its regulation.Results: YSSXL showed obvious therapeutic effect in treating β-thalassemia. Gene analysis revealed that it did not change the genetic mutation type, but could obviously increase hemoglobin, fetal hemoglobin (HbF), γ/(β+γ) globin ratio, γ-globin mRNA expression and GM-CSF mRNA expression in patients, as well as the GM-CSFmRMA in marrow of mice after60Co radiation.Conclusion: YSSXL has a remarkable therapeutic effect on β-thalassemia, and its possible mechanism is its action in unlocking γ-gene, increasing the γ-globin expression and enhancing HbF synthesis so as to compensate for the gene defect. This study has opened a new path for the treatment of β-thalassemia with TCM.

Key Words  nourishing Shen and supplementing marrow principle - Shen producing marrow - β-thalassemia - gene analysis - mRNA gene expression - molecular mechanism
This item was supported by National Funds of Natural Sciences (No. 30171199) and Natural Science Foundation of Guangxi Autonomous Region (No. 014402C)

[Effect on expression of mice alpha-hemoglobin stabilizing protein in different developmental stages treated with Yisui Shengxue granules]
[Article in Chinese]

Liu YM, Wu ZK, Chai LM, Zhang XH, Li M, Chen YY, Lv XX, Zhu XY.

Department of Biology, Guang' anmen Hospital, China Academy of Chinese Medical Sciences, Beijing 100053, China.

OBJECTIVE: To discuss the effect of Yisui Shengxue granules on expression of alpha-hemoglobin stabilizing protein (AHSP) mRNA in different developmental stages mice. METHOD: The total RNAs were extracted from the bone marrow karyocyte of normal adult mice and the karyocyte of fetus liver and fetus spleen in pregnanted mice (pregnanted 21 days) and fetal mice (pregnanted 14 days). The expression level of AHSP mRNA in different developmental stages mice interfered with Yisui Shengxue granules was measured by real-time PCR. RESULT: The intervention of Yisui Shengxue granules could significantly up-regulated the expression levels of AHSP mRNA in normal adult mice. CONCLUSION: The result revealed that one of possible molecular mechanism of the effects caused by Yisui Shengxue granules is that it can promote the AHSP gene expression, reduce the free a-globin deposit, then prevent the poison to erythrocyte and decrease the haemolysis.

A ray of hope for special people like my cutiepie that she'll be cured one day   
Apparently Yisui Shenxueling Granules are a traditional Chinese medicine, but so far, the only things I have learned about his is from these articles. If anyone knows someone in China who may be familiar with this, please make some inquiries.

Even if this is largely lost on the medical profession, common sense tells us that through the ages, there must have been many attempts to treat thalassemia with natural remedies. Unfortunately, as we "modernize", natural treatments are often forgotten and lost, so we know very little about whether any of these methods met with success. Investigations into traditional medicine may find treatments and even cures for many ailments, but how do we find out what these traditional medicines are now that we are so far removed from the past and the cultures that developed these medicines? I really think it shouldn't be left to only the pharma giants to pursue this knowledge.