You’ve read about the biotechnology in the use of yeast to create a product like bread, but yeast is not the only organism that can be used to create something. In fact, the very first human insulin produced in a lab in 1978, used E. coli – to the dismay of readers whose only knowledge of E. coli is through news reports of foodborne illness – as an essential part of the process. The key to the production of human insulin by bacteria is recombinant DNA – DNA from one organism that is “recombined” in the DNA of another.

Who & Why?

The firm that produced the first human insulin is Genentech (now a member of the Roche Group). In fact, Genentech is generally credited with being the very first biotechnology firm. The founders, Boyd and Swanson, decided that the way to build a successful company was to produce a product with large commercial potential. From the 1920s and up until Genentech’s success, insulin for human use was manufactured from the pancreas of slaughtered cows and pigs. It worked, but over time, many diabetics developed an allergic sensitivity to the animal insulin they received. If human insulin could be produced, the logic went, then that would eliminate the potential for an allergic reaction. Human insulin for human use would be the solution to a serious problem – and the market for human insulin at that time was large and growing. If they could just …

What?

Researchers decided to create a human gene for insulin and insert it into the genetic material of a bacterium. Once the genetic material was inserted into the bacterium, it was considered a “recombinant bacterium.” They selected E. coli as the bacteria. (Why? That’s another post.) Successfully creating the recombinant bacterium was the key to it all. To accomplish this in the lab, researchers:

1. created a human insulin gene.

2. removed a plasmid – a loop of DNA – from the bacteria.

3. inserted the human insulin gene into the loop.

4. returned the plasmid to the bacterium.

The DNA in the bacterium now contained material from humans and bacteria. It contained recombinant DNA.

How?

The recombinant bacteria are placed in a fermentation tank and grown under optimal conditions. Since part of their “job” is to produce human insulin – because it is now part of their (recombinant) DNA – they produce human insulin. All that is left is to harvest and purify the insulin. bio = E. coli bacteria + technology = Recombinant DNA. Biotechnology!

The U.S. National Library Medicine at NIH video below, “How did they make insulin from recombinant DNA?, illustrates the process described above.

Written for and originally published on AGT

Big Picture 

Gene therapy, stem cell therapy, CAR-T, cell therapy and gene editing are all forms of genomic medicine¹ – an approach to cure and treat human diseases that uses human biology rather than chemical compounds made in the lab. All of these tools are unlocking techniques and therapies with the power to cure formerly incurable diseases. Their use is ushering in a new era of healthcare.

What Is Gene Therapy? 

Gene therapy is one of the tools being used by scientists as a result of the mapping of the human genome and the advances made in biotechnology. Gene therapy uses genetic material to halt the progression of a disease or cure a genetic defect. But inserting a gene is not as simple as swallowing a pill or receiving an injection. In contrast with previous generations of medicine, gene therapy takes the genetic material required to treat a disorder and introduces it into the body for uptake into the appropriate cells through what is best described as a viral update. Here’s how it’s done.

How Does It Work?

Genes are the part of an individual’s genetic material that decides an organism’s features, how it behaves in its environment and how it survives. They hold the information to build and maintain an organism’s cells and pass genetic traits to the next generation. Genes consist of a long combination of four different nucleotide bases that are known as adenine (A), cytosine (C), guanine (G), and thymine (T). The most important thing about genes is that all living things depend upon this code working correctly.²

In a monogenic disease, there is an error in one gene which generates too much, too little  or incorrect protein. Most of these errors are present when we are born and populate every cell in our body. These errors can cause serious disease; however, if the error were to be corrected, it would make all the difference. So, suppose you have a gene with a mutation that tells a cell to make too much of a specific protein. In this case, the gene will continually overproduce which in some cases may not matter.  However If the product of that gene and protein is toxic in large quantities, it can cause a problem.

With underexpression of a gene, it raises a similar problem and we need to bring in outside help to solve it.  There are several ways to increase the product of a gene with another. The simplest way to increase expression would be to add a new copy of that gene which expresses independently normalizing the product of the gene. That’s where viral vectors come into play.

Viral Vectors — DoorDash for Genes 

In gene therapy, a viral vector can be a delivery mechanism for genetic material.³ In the case of a monogenic disease, the vector will deliver the material required to fix the damaged gene. It’s not possible to just walk up, knock on the door, and hand over the new gene. It is necessary for the gene to travel through your blood or intracellular space and find the right cells which need genetic improvement. Luckily viruses have been doing this for millions of years. So scientists have modified viruses to deliver genes to the appropriate “doors”.

There are many types of viral vectors like adeno associated virus (AAV), Lentivirus, etc, but the basic theory applied is always the same. Identify a virus,, remove all the pathogenic parts, especially the ability to replicate, and add the gene you want to deliver. Then allow the viral vector to do what viruses do in nature; deliver the carried DNA sequence. If we deliberately invade a cell with the new gene, that gene will enter the nucleus of the healthy cell and replace the function of the previously missing or broken gene. The net result? Improvement if not a functional cure!

Is it “Fixed” for the Next Generation?  

Can a genetic cure carry on to our children?  Today, that is not possible because gene therapy uses somatic cells – cells that are not part of the egg and sperm, also known as the germline cells. It is the germline cells that pass the gene to the next generation. Gene therapy now being used to provide a “fix” that impacts only the individual receiving the inserted gene. As you’ll read in the article on gene editing, it may be possible in the future to “fix” not only the defective gene in the target patient, but also in any future offspring as well.

Sources:

¹Uncovering the Relationship Between Genes and Proteins

²National Human Genome Research Institute – Vector

One straightforward definition of biotechnology I really like is that it is technology that utilizes biological systems, living organisms or parts of this to develop or create different products. It’s the definition I use because it is uncomplicated and sticks to the basics. Unfortunately, since people already realize that the term “biotechnology” is made up of bio + technology, this definition does not answer the question people are really asking. They invariably want to know two things: What is biotechnology used for? How does it work? Let’s keep it simple and start with bread.

What? We use biotechnology when we make leavened bread.

How? We use biotechnology by taking advantage of the natural activity of living organisms or parts of biological systems to reach our goal. Think about it: Yeast is a living organism. You can see that for yourself if the water you add at the start is too hot. The yeast will die because it does best under conditions that provide the ideal environment for its growth. Yeast doesn’t set out to turn flour into bread. It sets out to thrive and feed itself. During the process of fermentation, the yeast feeds on the sugars in the flour and expels carbon dioxide. Bubbles of carbon dioxide become caught by the gluten in the dough and the bread rises. In this instance, bio = Yeast + technology = Kneading/Resting/Baking. Biotechnology!

Sanjeev Luther, CEO and Board Member of Rafael Pharmaceuticals, pictured above. 

BioSpace recently spoke with Sanjeev Luther, CEO and board member of Rafael Pharmaceuticals, a clinical-late stage metabolic therapeutics company. Rafael’s lead drug CPI-613® (devimistat), from their Altered Metabolism Directed (AMD) platform, targets enzymes that are involved in cancer cell energy metabolism and are located in the mitochondria of cancer cells. Rafael also has an interest in ultra-orphan and orphan drugs because the need is so great in this area.

Q: What are your daily activities as CEO of Rafael?

A: I’m involved in the administrative aspect of my position maybe 25% of the time. The other 75%, I’m actively involved in the operations of the company. Rafael has gone from four or five people at the start to ten or eleven employees when we started our large trials in 2011. Because of this, I visited every single site we opened, whether it was in the U.S. or globally, in the first six months. Now we have fifty employees, but as a hands-on operations guy, I’m used to rolling up my sleeves and just doing it.

Q: How did you choose to work in the biopharma industry?

A: I picked pharma. I went from my MBA program to a consulting firm in New York City. I had many different clients from a variety of industries. The work was interesting, but I fell in love with pharma for a very simple reason: It was a more strategic, longer-term outlook than the other industries I worked in. And this right here was more tangible to me. I felt like I could hold something. I can be part of something. It gave me a lot of excitement.

Q: Tell me about the work Rafael is doing with pancreatic cancer.

A: We have CPI-613 in trial as a treatment for pancreatic cancer. It’s an immunologically cold tumor. Our focus is on cold tumors both because immunotherapy does not work on these types of tumors and there is a great need for treatment because of the mortality rate.

CPI-613 is a small-molecule, agnostic metabolic treatment, meaning it will go to a cold tumor and deprive it of nutrients. The idea is then to use less chemo than is the standard of care to kill the tumor. If this proves impossible, Rafael will set its sights on the next generation of CPI-613 and/or two other possible strategies. Our interest is in ultra-orphan and orphan drugs because the need is so great and, since it is not a large market, many companies do not invest in this area.

Q: What is the best part of your work?

A: Patients and hope. I feel this glimmer of happiness. I’m very well connected with all physicians who work with us and all the patients – of course, there’s HIPAA so I don’t directly go to the patient. But any caregiver or patient can call me directly at any time. So that’s what I enjoy the most: the next indication, next patient, next physician, and the next thought.

Q: Anything else you’d like to add?

A: We just got American Cancer Society data this week. Every cancer has declined except pancreatic cancer. The newest data is very interesting. It says that every day in the U.S., 166 people will find out that they have pancreatic cancer. That’s a huge number.

Last year, we used the number that 55,000 people would get pancreatic cancer. It is the only cancer that was updated – to 62,000 this year. And the five-year survival went down. So it used to be 12%, five-year survival, they cut it down to 10%. Double whammy, right? More people getting it, and fewer people living. So we’re very dedicated to our work on pancreatic cancer. It’s hard; in the last 14 years, no drug has been approved for pancreatic cancer. The drugs that are there, they’re chemo drugs basically, and they don’t work very well. Our ultimate goal is to eventually come to where there’ll only be a cancer metabolism drug, and you won’t have to use the chemo as a combo. That’s where our focus is right now.

Published on BioSpace

There’s a lot of hype around biotechnology and its potential to change the world – for good or bad. Most of the hype is true. The right biological materials in the wrong hands can lead to disastrous consequences for us all. Then again, the right materials in the right hands … You get the picture. So, is biotech good or bad? And, for that matter, what is biotech? To answer those questions, you need to be comfortable with some basic concepts and applications. Once you have those, you are, as they say, good-to-go.

For the next few weeks, I’ll meet you here to present a series of posts – Biotech 101. I’ll give you the scoop on the different “types” of biotech, along with all the basics about this amazing blend of biology and technology. I’ll give you that scoop without jargon or complicated explanations. No less than Nobel Prize winner Richard Feynman said that if you couldn’t explain it simply, you didn’t understand it. So, let’s accept that as our premise and see how far we can go.

I’ll meet you back here next week to get this conversation started!

See you then!

~ gina

There are 7,000 known rare diseases in the world. Each of them affects less than 1% of the global population, although ultra-rare diseases affect fewer still. The majority of rare and ultra-rare diseases are monogenic in nature, meaning they are caused by an error in a single gene. Years ago, a rare disease diagnosis left parents with their worst fears realized and little that could be done. Today, genomic testing can pinpoint the exact location of the genetic error, while gene therapy presents the possibility of therapy for the specific disorder – therapy that may prove to be a cure. Even with genetic testing and gene therapy though, it is not a “done deal” that a child will receive gene therapy after diagnosis. The three “Rare Moms” included here are some of the many who are pushing hard for the development of treatments for their children and others like them.

1 – Amber for Maxwell | SLC-6A1 

Amber Freed’s son, Maxwell, was just a few months old when Amber realized he was not progressing at the same rate as his twin sister, Riley. As is common, it took several months before she began her diagnostic odyssey, going from specialist to specialist in search of a diagnosis. Her path ultimately led her to a geneticist who would analyze Maxwell’s genome to discover the cause of his disorder. Four months later, she had the answer: SLC-6A1. This was the location of the mutated gene. The disease itself was so rare it had no name.

Amber’s efforts to find a researcher familiar with the disorder, raise funds for the necessary studies, organize a clinical trial, and more for Maxwell led her to found SLC-6A1 Connect. This nonprofit serves as a source of news, support, and fundraising for the millions of dollars needed to cure her son and others who are diagnosed with SLC-6A1. Amber says of the money required, “Like any new technology, the cost is astronomical today but will decline as the process becomes more automated. However, Maxwell does not have the luxury of time, so the money needs to be raised now.” Every day, she works to raise awareness and funds to continue research to help Maxwell and others like him.

2 – Alison for Tia | Crossing Norway PKU 

Alison Reynold’s daughter, Tia, was two days old when Alison received a call that changed their lives. Tia’s routine infant screening panel revealed she had phenylketonuria (PKU), a rare metabolic disease. This rare disorder results in severe intellectual disability unless a very strict dietary regimen is followed – for life. Since this dietary regime was created and tested by Dr. Asbjørn Følling in Norway more than 80 years ago, it has been the only proven expedient for those with PKU. In her teens now, Tia has begun taking a new medication called Palynziq to lower the levels of Phe (phenylalanine) in her blood to help to prevent it from building to unhealthy levels. Since Palynzig came on the market in 2018, it will take some time before its long-term effect is apparent.

In 2019, Alison decided that more was needed to raise awareness about PKU and celebrate the work of Dr. Følling. After months of training and preparation, this Washington, D.C. mother of four, began her Crossing Norway for a Cure trek along the 125-mile Norway/Sweden border, just south of the Arctic Circle. Her cross-country trip included nine-hour days of trekking for nine days, hauling her equipment on an 80-pound pulk sled by day, and sleeping in a tent on the snow at night. Alison and her guide, Elise Koren, finished their trek on Feb 29, 2020, having raised $1.7 million for PKU awareness. Alison is hoping a cure will be discovered during Tia’s lifetime. In a 2019 interview with Washingtonian magazine, Alison said, “It’s a hard road. Every morning you’re reminded of the quest you’re on. We’re on a mission, and I think we’ll get there.”

3 – Anne for Nick | Partners for Krabbe Research

Anne Rugari’s second son, Nick, was diagnosed with Krabbe disease, a rare genetic disorder that attacks the myelin-protecting nerve cells in the brain and nervous system when he was seven months old. Without treatment for his stage of illness, Nick died just three days after his first birthday. When Anne was unexpectedly pregnant with her second child, she knew that early diagnosis was essential. With a diagnosis in hand before symptoms began, her daughter, Gina, received chemotherapy drugs before she was four months old so that she could get an experimental cord blood transplant. Gina is one of four infants in the world with Krabbe disease to receive a transplant as an infant. She lived for fourteen years longer than Nick. Many doctors and researchers studied and monitored her progress for insight into future treatments for Krabbe disease.

Anne founded Partners for Krabbe Research and is the co-founder and Vice President of KrabbeConnect. She shares her experience with others and has written two books for kids. In addition to donating Gina’s brain and tissue to the NORD Program at the University of Pittsburgh Medical Center, she works with those searching for a cure. Anne says, “I have been given the opportunity to work with researchers, clinicians, scientists, and families through the 33 years since my son Nick was diagnosed.” She added that her goal is “to be a voice as an advocate for those affected by Krabbe disease by making a difference through education, research, and awareness.”

Takeaway

Amber Freed, Alison Reynolds, and Anne Rugari each took action in response to the rare disease diagnoses of their children. Establishing an organization, building awareness, and raising funds for rare diseases are the ways they chose to help their children and others like them. By putting a face to a little-known disorder, they are encouraging those outside the rare disease community to support the development of treatments for those with rare and ultra-rare diseases. They are also spotlighting the need for continued investment, government support, and the support of those like AGT in the biotech community to find cures or treatments that will help those with rare and ultra-rare diseases.

Want to help?

Support these moms:

• Amber Freed – SLC-6A1/Milestones for Maxwell
• Alison Reynolds – PKU/NPKUA
• Anne Rugari – Krabbe/Partners for Krabbe Research

Learn more about rare diseases:

• NORD (National Organization for Rare Diseases)

Published on AGT