Friday, September 19, 2014

New Haven Register article on Dr. Crawford's Patent Award, November 29, 2013

By Jim Shelton, New Haven Register
POSTED: 11/29/13, 2:07 PM EST | UPDATED: ON 11/29/2013

New Haven >> In the little laboratory she built from scratch, Sarah Crawford is working on a Christmas fern miracle.

Not that she’d ever use the word “miracle” to describe the cancer research she’s been conducting at Southern Connecticut State University for the past 17 years. She’s much too measured for that. Besides, she says the media seems to tout a new cure for cancer every other week.

Yet, it’s undeniable what Crawford has accomplished outside the realm of high profile, big money cancer research. Not only is she rethinking basic assumptions about the disease, she recently was granted a patent for a Christmas fern extract that shows potential for treating brain cancers.
The extract, used in a cocktail with the chemotherapy drug carmustine and curcumin, the active ingredient in turmeric, killed nearly half of the cancer cells in animal tumors during pre-clinical testing. Crawford said that within a year, she’ll be ready to approach pharmaceutical companies and large research labs about doing a human study.

Not bad for a college professor without major funding or a staff to write grant proposals.

“There needs to be a place for the individual researcher in science,” Crawford said. “You have to be able to close a door, sit down and think something through. But you also have to be willing to do without the money and be able to improvise.”

In her case, that means going to state parks on the weekends to gather Christmas ferns and using chicken eggs to test tumor treatments.

Actually, it took a fair bit of gumption just to envision her lab at SCSU in the first place. She’d come to the university in 1993 to teach genetics, raise her two young daughters and figure out what sort of cancer research she’d like to do elsewhere.

But then she got to thinking.
“I walked into this sort of broken down room, and the thought occurred to me, ‘You could have your own lab right here. Do the research you want to do.’” The lab opened — with no fanfare — in 1996.

Her focus, she decided, would be on drug resistance. Why don’t certain cancers respond well to treatment? Is it because we treat cancer only as an acute disease rather than a systemic one?

Crawford began studying the use of aspirin and anti-inflammatory drugs on tumors. She found that solid tumors responded slowly to large amounts of aspirin.

Then she broadened her approach and looked at plant products with high antioxidant activity. That’s how she arrived at Christmas ferns.

“They’re primitive plants,” Crawford said. “They have a very different biochemical composition. These plants were some of the first to evolve, and they’re very successful.”

She and her students started very simply, grinding up ferns and looking for the key ingredients. They discovered that in concert with curcumin and carmustine, the ferns produced far better cancer-fighting results on tumors than when any of the three substances was used alone.

Crawford applied for a patent more than six years ago; she now shares it with Erin Boisvert, one of her former students.

Of course, this is only the beginning of the process for Crawford. She has plans to tinker with the drug cocktail and test it on pancreatic tumors.

“I always temper my excitement with reality,” she said. “I’m not going to rest until we, as a research community, are able to effectively deal with this disease. We have to find a cure.”

Crawford’s motivation stems in part from a childhood experience.

At the age of 9, she spent two weeks in the children’s ward of a New York City hospital. It was Christmas time, and Crawford met children dealing with cancer, kidney disease and other dire ailments. One child died while Crawford was on the ward.

“It was one of the major experiences of my childhood. It changed my life,” Crawford said. “I’ll never forget walking out of there and knowing I would be studying disease for the rest of my life.”

By now, more than 100 students have come through Crawford’s little lab. In 2015, she’ll move to a brand new building on campus, the Academic and Laboratory Science Building.

But don’t expect her to take on expensive research habits.

“It certainly has its place, but that focus on funding keeps you thinking inside the box,” Crawford said.

European Conference of Oncology Pharmacy, 2014 Presentation:
A new preclinical Human Cancer Model demonstrates successful Growth of Human Solid
Tumors of diverse Tissue Origins in unfertilized Avian Eggs – Dr. Sarah Crawford (New
Haven, USA) P3

Cold Spring Harbor Meeting: "Avian Model Systems" March 2014 Presentation by Dr. Crawford

A new preclinical human cancer model demonstrates successful growth of human solid tumors of diverse tissue origins in unfertilized avian eggs


A novel pre-clinical approach developed in this laboratory for the first time demonstrates the successful growth of human solid tumors of the brain, breast, colon and pancreas in unfertilized chick eggs*. The research suggests that the avian system is a suitable and in many ways more advantageous culture system for the growth of human tumors than current in vitro and mouse model systems.

Growth advantages offered by the avian system include:
• High success transfer rate (at least 80%) from in vitro spheroid culture to avian culture.
• Rapid establishment within days of solid tumor growth.
• Broad spectrum application to tumors of diverse tumor type grown successfully in this system, including tumor spheroids of the brain (glioblastoma), pancreas (ductal adenocarcinoma), colon, and breast (primary and metastatic origins).
• Capacity for long-term cultivation not possible in fertilized chick eggs by serial transfer.
• Avian embryonic environment affords biochemical , biophysical parameters and determinants of growth properties difficult to achieve in adult animal models or in vitro systems.
• Ease of culture manipulation and tumor growth assessment.
• Tumor growth in avian eggs displays heterogeneous growth parameters more similar to in vivo growth than spheroid cultures, including clear distinctions between necrotic tumor centers and active growth zones, increased invasiveness compared to traditional spheroid growth models and histological heterogeneity similar to that observed in human tumor specimens observed surgically and histologically.

In summary, research demonstrating the simplicity, efficacy and greater similarity in tumor growth properties in the avian system as compared to in vitro spheroid culture methods will be presented, including data showing the use of this model system to assess the potential efficacy of novel therapeutic approaches and as a comparative tool for assessing pre-clinical versus patient treatment responses.

*Provisional patent application, Sarah Crawford, 2014.

Wednesday, November 13, 2013

Patent Award for Brain Cancer Rx


sarah crawfordChristmas ferns are pretty ordinary plants in the botanical world – they are in abundance in the eastern half of the United States, and if you walk through a shaded area of a park, you can find them pretty easily.
Yet the properties inside this ostensibly unremarkable fern may be a catalyst toward combatting an aggressive form of brain cancer. So says, Sarah Crawford (left), a professor of biology at Southern who has an extensive background in cancer research.
But her statement is more than just an abstract theoretical possibility. An extract made from the Christmas fern has demonstrated anti-cancer properties in pre-clinical testing conducted by Crawford and her students. In fact, the results were impressive enough to spur the U.S. Patent Office to award Crawford, as well as Erin Boisvert, a former student of Crawford, a patent for the extract.
"This is really exciting news," Crawford says. "I applied for the patent more than six years ago and was hopeful it would grant its approval. But it's a long, thorough process. You're never quite sure whether it is going to be approved or not."
The extract was tested as part of a three-component cocktail – carmustine, a powerful chemotherapy drug used to treat brain cancer; curcumin, the active ingredient in the spice turmeric that has anti-inflammatory qualities; and polystichum acrostichoides, the technical name for the Christmas fern. The plant is believed to have antioxidant properties, but to Crawford's knowledge, it has not previously undergone rigorous testing for its anti-cancer ability.
cancer researchThe tests showed that the cocktail was effective in killing nearly half of the cancer cells in tiny tumors created in the Biology Department lab – far more effective than use of any of the three substances alone. "I won't rest until we can kill 100 percent of the cancer cells, but it's a good start," she says, adding that she plans to experiment by using different levels of each substance to see if that increases the efficacy of the extract's anti-cancer properties. She said she also may test other chemotherapy drugs with the Christmas fern and curcumin.
Crawford says that a reduction in the level of carmustine, but maintaining or increasing the effectiveness of the cocktail, would be ideal. That could reduce the side effects commonly associated with chemotherapy drugs.
The tests were conducted on glioblastoma multiforme, considered to be the most deadly form of brain cancer with a fatality rate of more than 90 percent within five years.
Two current students are assisting Crawford with this project.
Brielle Hayward, who is a graduate fellow, is examining the Christmas fern's antioxidant properties and comparing its anti-cancer effects with other phytochemical antioxidants, such as American and Korean ginseng.
Paulina Mrowiec, who is a member of the Honors College, is continuing her research on the project after completing an undergraduate thesis last spring on pre-clinical models for cancer drug testing.
Crawford says she looks forward to the opening of the Academic and Laboratory Science Building, scheduled for 2015, which promises state-of-the-art facilities and equipment to conduct further research.


511 ‘Ex vivo’ culture of human micro-tumors in unfertilised avian eggs
Avian eggs, for tumour culturing / Pre-clinical models / Tumour culturing, in avian eggs
S. Crawford P. Mrowiec, K. Floyd Southern Connecticut State University, Biology, New Haven Connecticut USA
Background: The quest for more relevant pre-clinical models for testing cancer drug efficacy is a primary focus of cancer research. Current in vitro protocols are poor representations of the cancer microenvironment inside the body. The focus of this research study was to evaluate a novel pre-clinical model involving the culture of tumor cells in unfertilised avian eggs. The choice of this experimental system is based on the hypothesis that the avian egg may represent a pre-clinical model of intermediate complexity between the in vitro system and the in vivo setting that combines essential elements of the in vivo system with the simplicity of a highly accessible, stable and quantifiable 'ex vivo' system to assess tumor biology and drug sensitivity. Fertilised avian eggs inoculated with human tumor cells have been used to assess tumor neo-angiogenesis of the chorioallantoic membrane; to date, there are no reported studies of tumor cells cultured in unfertilised avian eggs.
Materials and Methods: Unfertilised avian eggs were inoculated with tumor cells from glioblastoma and juvenile osteosarcoma cell lines in the following procedure: a 1–2 cm opening in the egg shell and membrane was made by small punctured hole on top of the egg. Egg white (albumen) was removed with a syringe and different amounts of media were replaced. Trypsinised tumor cells or tumor spheroids from suspension culture were inoculated into the albumin portion of the egg. Eggs were covered with sterile gauze pads and placed into the incubator where they were incubated at 37°C with a atmospheric saturation of 5–10% CO2 from 4 days to 2 weeks. To assay tumor growth, 1 ml of incubated albumin contents were removed with a syringe and placed onto a monolayer culture plate containing standard culture media. After 24 hours the egg contents were observed. Tumor attachment to culture dish substrate and the trypan blue exclusion assay were used as indicators of viability.
Results: Live cell photo microscopy studies showed that both cell lines were capable of proliferation and micro-tumor formation in the albumin component of unfertilised avian eggs when at least 15–20% of the egg albumen was replaced by complete culture medium (RPMI 1640 or DMEM) supplemented with 10% serum. When cells inoculated into avian eggs were transferred to liquid culture, cells were able to reattach and form small cell masses indicative of micro-tumor formation.
Conclusion: This research has shown that two tumor cell lines of diverse tissue type can be successfully cultured in unfertilised avian eggs. This may be the first series of experiments to show the possible use of this system to support the growth of tumor cells. Future studies will attempt to quantify the growth parameters, biological properties and treatment sensitivity of tumor cell lines cultivate in this novel ‘ex vivo’system.