Journal Of Virology Reports On Link Between HIV and Niemann Pick Cholesterol Gene: Intact Intracellular Cholesterol Trafficking Pathways Mediated by NPC1 are Needed for Efficient HIV-1 Production

Journal of Virology
May 27, 2009

Deficiency of Niemann-Pick type C-1 Protein Impairs HIV-1 Release and Results in Gag Accumulation in Late Endosomal/Lysosomal Compartments

Tang Y, Leao IC, Coleman EM, Broughton RS, Hildreth JE.

Human immunodeficiency virus type 1 (HIV-1) relies on cholesterol-laden lipid raft membrane microdomains for entry into and egress out of susceptible cells. In the present study, we examine the need for intracellular cholesterol trafficking pathways with respect to HIV-1 biogenesis using Niemann-Pick type C1 deficient cells (NPCD) wherein these pathways are severely compromised, causing massive accumulation of cholesterol in late endosomal/lysosomal (LE/L) compartments.

We have found that induction of an NPC disease-like phenotype through treatment of various cell types with the commonly-used hydrophobic amine drug U18666A resulted in profound suppression of HIV-1 release.

Further, NPCD EBV-transformed B lymphocytes and fibroblasts from patients with NPC disease infected with a CD4-independent strain of HIV-1 or transfected with an HIV-1 proviral clone, respectively, replicated HIV-1 poorly when compared to normal cells.

Infection of the NPCD fibroblasts with a VSV-G pseudotyped strain of HIV-1 produced similar results, suggesting a post-entry block to HIV-1 replication in these cells. Examination of these cells using confocal microscopy showed an accumulation and stabilization of Gag in LE/L compartments. Additionally, normal HIV-1 production could be restored in NPCD cells upon expression of a functional NPC1 protein, and overexpression of NPC1 increased HIV-1 release.

Taken together, our findings demonstrate that intact intracellular cholesterol trafficking pathways mediated by NPC1 are needed for efficient HIV-1 production.

The Wall Street Journal Reports On FDA Approval of Addi and Cassi’s Cyclodextrin Treatment

By AMY DOCKSER MARCUS
April 3, 2009
A Mom Brokers Treatment for Her Twins’ Fatal Illness Bucking Scientific Convention, Ms. Hempel Gets Researchers From Different Fields to Share Data on Potential Therapy

From the moment her twin daughters, Addison and Cassidy, were diagnosed with a fatal genetic disease in October 2007, Chris Hempel has been searching for a drug that might save their lives.  The 5-year-old girls were diagnosed with a devastating cholesterol metabolism disorder known as Niemann-Pick Type C, which is ultimately fatal. Soon after, Ms. Hempel learned that researchers found that a form of a compound called cyclodextrin extended the lives of affected mice. Ms. Hempel set out to gather as much data as possible. She got a list of all major cyclodextrin distributors and connected with one in Florida, who shared scientific studies and other information with her. She found a short report in the medical literature about a doctor who had treated a child with a different disease using cyclodextrin and tracked him down. She became increasingly hopeful that, although cyclodextrin isn’t approved as a drug in the U.S., she might get the Food and Drug Administration to allow her to give cyclodextrin infusions to her girls as an experimental treatment.

Her search for information also led her to James Hildreth, 52, a pre-eminent AIDS researcher who heads the Center for AIDS Health Disparities Research at Meharry Medical College in Nashville, Tenn. It turned out that he too was seeking FDA approval to run a trial using cyclodextrin, in a vaginal cream to help prevent HIV transmission during heterosexual sex. Ms. Hempel wanted him to combine forces with the NP-C investigators to push forward cyclodextrin research.

That was only the beginning of Ms. Hempel’s long journey through the health-care research community — a distributed and labyrinthine collection of researchers who, for all their expertise, often remain unaware of advances made elsewhere. The problem is even more acute among researchers working on different diseases. But for some serendipity, curiosity — or, in this case, a willful Ms. Hempel — some knowledge in one lab may never make its way to another that could be on the verge of a new therapy.

Drugs approved for one disease often turn out to be effective in others — frequently when someone has a hunch. Thalidomide, originally used for morning sickness but taken off the market because it caused birth defects, is being used in cancer treatment. Researchers at Pfizer were developing Viagra to treat high blood pressure when they noticed during early tests that it treated impotence. But that happened within the same company. It is even more difficult when researchers are at different labs.

When Ms. Hempel, who lives in Reno, Nev., became passionate about Dr. Hildreth’s work, she was determined to bridge the disparate knowledge. "Right now we have limited data on cyclodextrin. But what if a lot of people started looking at it from different angles and across different diseases?" Ms. Hempel said. "It could lead to something that helps save Addi and Cassi’s lives."

Ms. Hempel had been researching cyclodextrin for months when she attended the June 2008 meeting in Tucson, Ariz., of the Ara Parseghian Medical Research Foundation, set up by the family of the legendary football coach who lost three grandchildren to NP-C disease. The foundation was providing some funding for cyclodextrin studies in the rare disease, and the latest data were presented there. In an email sent after the meeting, Ms. Hempel wrote to the NP-C researchers that, based on the data she heard, she and her husband, Hugh, planned to seek FDA approval to give the girls cyclodextrin infusions. "I feel very strongly that we must try this to help save Addi and Cassi from this horrible disease," she wrote. She had already put together a three-inch binder of research studies about cyclodextrin. Working with three other families whose children have NP-C disease, they hired a scientist who began writing a request to the FDA for the Hempel children to receive cyclodextrin infusions.

But Ms. Hempel knew that she needed more human data if she was going to persuade the FDA that the drug was safe enough to use in her children. While searching for safety data on cyclodextrin, she spoke with Charles E. Strattan, a cyclodextrin expert and CEO of CTD Holdings Inc., who was helping Ms. Hempel do research. He told her Dr. Hildreth was interested in the same compound for his work in HIV and suggested that the two of them talk.

During a long phone conversation in October 2008, Dr. Hildreth told Ms. Hempel that he believed the protein responsible for NP-C disease also plays an important role in HIV. And in previously published work, he showed that cyclodextrin appeared to inactivate the HIV virus and prevent it from replicating. The talk galvanized Ms. Hempel. Dr. Hildreth offered to share what he knew about cyclodextrin’s safety with the FDA in support of the Hempels’ request. Ms. Hempel proposed that the two of them go to Johnson & Johnson, which had studied cyclodextrin, to see if the company would be interested in sponsoring a clinical trial. "I knew our stories would be even more powerful if we told them together," she said.

As is typical in the field, Dr. Hildreth was reluctant to share unpublished data, and he rarely went to scientific meetings that weren’t related to HIV. He was moved by Ms. Hempel’s efforts to help her children, but also surprised by her embrace of his work. "Some of the things we as scientists take for granted about how work will be done and the fact there are silos, with her there is none of that at all," he said.

When Ms. Hempel called a top National Institutes of Health AIDS researcher to tell him about Dr. Hildreth’s findings and propose joint work in HIV and NP-C disease, Dr. Hildreth told her that a scientist never would have made such a call. In recent months, Ms. Hempel has introduced Dr. Hildreth to NP-C researchers who were also studying cyclodextrin. She also arranged for him to discuss his HIV findings with two Nobel Prize-winning scientists interested in Niemann-Pick proteins. "Our paths would not have crossed otherwise," he said.

He and Ms. Hempel recently had a conversation with senior officials at Johnson & Johnson. The FDA at first turned down the Hempels’ request to do cyclodextrin infusions in the girls, concerned there wasn’t enough human safety data. But after Ms. Hempel contacted them about her plight, the company wrote a letter to the FDA giving the agency permission to look at all of the safety data it had submitted related to cyclodextrin. The FDA subsequently gave permission for the Hempels to proceed. The girls will start cyclodextrin infusions this month.

That might have been the end of the story except for Ms. Hempel’s insistence that more was at stake, says Steven A. Silber, a vice president at Johnson & Johnson. After listening to Ms. Hempel and Dr. Hildreth’s presentation, Dr. Silber set up a meeting so Dr. Hildreth can present his data to the head of one of its companies that makes anti-viral medications. Dr. Hildreth says that Ms. Hempel’s involvement got his research "the attention of individuals higher up in the organization than I might have been able to get on my own."

This May, the Parseghian Foundation will host its annual scientific meeting. The group plans to hold a special session dedicated to the work on cyclodextrin. Cindy Parseghian, president of the foundation, says she hopes researchers working with cyclodextrin in other diseases will also attend. "We think there should be more cross-fertilization," she said. Dr. Hildreth says he plans to share his findings at the meeting. Dr. Hildreth recognizes that his unusual partnership with Ms. Hempel also has some risks for the HIV trial he is planning. "It is a remote possibility, but is a possibility, that if her beautiful girls are done some harm by the infusions, that would clearly do harm to our efforts," he said. Still, he adds, "I spent a lot of time thinking about what I would do if I were in her position. My answer is I would do exactly the same thing."

Late last month, the Hempel girls underwent surgery at a California hospital to get a small medical device implanted under their skin to make it easier to receive regular cyclodextrin infusions. Dr. Hildreth visited them in the hospital.

It’s Time Dr. Francis Collins Be Named Director of The National Institutes of Health

News agencies over the past month have reported that Dr. Francis S. Collins, the scientist who led the U.S. government drive to map the 3 billion letters of the human genetic code, is the leading contender to run the National Institutes of Health.  Where is the announcement we are all desperately waiting for?  God, what is the hold-up?

President Obama, if you want to overhaul our healthcare system like you say you do please give this critical healthcare post to Dr. Collins.  Dr. Collins understands that we can accelerate research by understanding our genetic code and how genetics influence our health and our lives.

Dr. Collins understands that studying genes that cause horrible rare diseases will aid in our understand the most common illnesses impacting millions.  For example, if you want to understand “obesity,” researchers need to understand how the Niemann Pick Type C gene on Chromosome 18 works.  The NPC gene regulates human cholesterol metabolism and has been recently linked to obesity and many other diseases that are affecting millions yet very little money at NIH goes into research in this critical cholesterol gene.

The focus at NIH needs to shift to the 23 pairs of chromosomes and the 20,000 genes that make us human and to understanding rare diseases.  We need Dr. Collins to continue to provide his revolutionary contributions to genetic research and continue to bring his intellectual and spiritual perspective to people everywhere.   I am not a scientist but I am a believer.   A believer in Dr. Francis Collins!

Obesity and Ordinary Weight Gain Linked To Guess What? Not Simply Junk Food But A Gene Called Niemann Pick Type C.

The first genetic map of obesity has been constructed using DNA microarray technology and guess what gene obesity is linked to?  The Niemann Pick Type C gene on Chromosome 18 that is mutated in Addi and Cassi and which is causing their deadly cholesterol metabolism disorder.   I have now found out that the NPC gene that is involved in all people’s cholesterol metabolism is liked to the HIV/AIDs viruses ability to assembly itself in the body of infected persons, Cystic Fibrosis, Muscular Dystrophy and now Obesity!

Would the NIH please provide more funding into studying the NPC gene — the NPC cholesterol gene dates back 500 million years to worms, flies and plants and obviously is involved in many critical processes in the human body.

The obesity report was publsihed in Nature Genetics by a research group led by CNRS researcher Philippe Froguel and Inserm researcher David Meyre.  This study led to the discovery of three new genes that increase the risk not only of severe obesity but also ordinary weight gain in the population.  It underlines that there is no difference between being overweight and other forms of obesity (mild, severe or massive).

Even though the increase in the number of obese people over the two last decades is partially due to social causes (inactivity, junk food, etc.), heredity plays an important part in determining body weight (70% hereditary) and the occurrence of obesity, especially when this is severe and appears early in life, according to researchers.

Froguel’s team has been working for 15 years to better understand the molecular basis of type 2 diabetes and the obesity found in 80% of diabetics.  Their work has revealed several genes responsible for monogenic forms of obesity and has demonstrated the essential role these genes play in appetite control.   The scientists first confirmed that the genes FTO and MC4R(4) played a major role in susceptibility to common obesity and weight gain in the population as a whole.  These two genes work by controlling eating behavior.

The researchers also found variations in the DNA close to the genes MAF and PTER(5), and directly in the coding sequence of the NPC1 gene.  Mutations associated with obesity could therefore directly induce an increase in the function of the NPC1 protein, such that it would work too well if the gene had mutated.

As for the MAF gene, it codes for a particular protein involved in the differentiation of adipose tissue (tissue responsible for fat storage) and in the production of a digestive hormone involved in satiety and insulin secretion.  The last gene (PRL) is more particularly associated with obesity and weight gain in adults.  PRL produces prolactin, a hormone well known for its effect in stimulating lactation in women.  Prolactin also plays a role in controlling the amount of food we consume.

Cyclodextrin As A Therapeutic “Drug” For HIV AIDS, Niemann Pick Type C and other Viruses

Addi and Cassi’s first round of cyclodextrin infusions have been going smoothly at Renown Regional Medical Center in Reno, Nevada.   We’re now into our third day of continuous infusions of hydroxy propel beta cyclodextrin (HPBCD) into the girls’ bloodstreams and they don’t seem to be experiencing any negative side effects.    I feel as if it’s having a positive and immediate effect.  Addi was talking yesterday afternoon and stringing together more than one word — “I like my toys, I’m brave enough, I need your help, “Bye” to Dr. Hastings, and “Ad” for her name Addison.  Even Cassi came out with a few words – “Mommy, No.”     This is quite encouraging to us but we’re not yet sure if it’s a result of the cyclodextrin treatment.

Addi and Cassi’s blood work-ups have come back “normal” following the cyclodextrin infusions.  There was a slight elevation in both girls’ eosinophils after 24 hours but it was minor.   We also had to change Addi’s port access needle as it was not working properly and we were unable to draw blood.   Unfortunately, we had to re-install the needle on her chest without any numbing cream.  Ouch!

I found out this morning that researchers were looking at the same cyclodextrin (HPBCD) and Niemann Pick Type C disease back in 1996 — 13 years ago!   I received this information from a scientist in Europe and I almost had a heart attack when I read the scientific abstract.

Somehow cyclodextrin research as it relates to Niemann Pick Type C was not thoroughly pursued with all angles exhausted by scientists.   This simply can not happen again — not only for Niemann Pick Type C disease but for HIV/AIDS and potentially other viruses like Herpes that are inactivated and killed by cyclodextrin.

I worry about the future of cyclodextrin research.  We can’t count on pharmaceutical companies to research cyclodextrin or bring therapeutic products with cyclodextrin to people as they are focused on profits and patents on new drugs that take millions of dollars and years to make.   Cyclodextrin is an inexpensive and non-toxic compound that can be deployed tomorrow — far too EASY!   But the HIV/AIDS pharma companies should be watching cyclodextrin very closely.  I believe the smart ones will start investing into research and try and create new patents around cyclodextrin since the Niemann Pick Type C cholesterol gene/protein on Chromosome 18 is the culprit in HIVs ability to assemble itself in the human body.

It’s time that the National Institutes of Health (NIH) Office of AIDS Research or the Centers For Disease Control and Prevention or some other government agency steps in and redirects money into cyclodextrin research to study this potentially life-saving compound that has very broad applications.

I am also asking for help from private citizens and global foundations such as The Gates Foundation, The Clinton Foundation, Elton John AIDS Foundation, amfAR and One.org to step in and help by taking a closer look at cyclodextrin and it’s relationship to cholesterol metabolism and killer viruses like HIV/AIDS.   We must make sure cyclodextrin is properly tested as a therapeutic agent whether money can be made from it or not (which is can be!)

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