Assessing the functional significance of novel putative oncogenes remains a significant challenge given the limitations of current loss-of-function tools. Here we describe a method that employs TALEN or CRISPR/Cas9 mediated knock-in of inducible degron tags (Degron-KI) that provides a versatile approach for the functional characterization of novel cancer genes and addresses many of the shortcomings of current tools. The Degron-KI system allows for highly specific, inducible, and allele-targeted inhibition of endogenous protein function, and the titrate-ability of this system is able to better mimic pharmacological inhibition compared to RNAi or genetic knockout approaches. The Degron-KI system was able to faithfully recapitulate the effects of pharmacological EZH2 and PI3K inhibitors in cancer cell lines. The application of this system to the study of a poorly understood putative oncogene, SF3B1, provided the first causal link between SF3B1 hotspot mutations and splicing alterations. Surprisingly, we found that SF3B1 mutant cells are not dependent upon the mutated allele for in vitro growth, but instead depend upon the function of the remaining wild-type alleles. Collectively, these results demonstrate the broad utility of the Degron-KI system for the functional characterization of cancer genes.

Cancer cells are surrounded by a fluid–mosaic membrane that provides a highly dynamic structural barrier with the microenvironment, communication filter and transport, receptor and enzyme platform. This structure forms because of the physical properties of its constituents, which can move laterally and selectively within the membrane plane and associate with similar or different constituents, forming specific, functional domains. Over the years, data have accumulated on the amounts, structures, and mobilities of membrane constituents after transformation and during progression and metastasis. More recent information has shown the importance of specialized membrane domains, such as lipid rafts, protein–lipid complexes, receptor complexes, invadopodia, and other cellular structures in the malignant process. In describing the macrostructure and dynamics of plasma membranes, membrane-associated cytoskeletal structures and extracellular matrix are also important, constraining the motion of membrane components and acting as traction points for cell motility. These associations may be altered in malignant cells, and probably also in surrounding normal cells, promoting invasion and metastatic colonization. In addition, components can be released from cells as secretory molecules, enzymes, receptors, large macromolecular complexes, membrane vesicles, and exosomes that can modify the microenvironment, provide specific cross-talk, and facilitate invasion, survival, and growth of malignant cells. Cancer Res; 75(7); 1–8. ©2015 AACR.

This symposium was held in Trento, Italy, on June 19–21, 2014, and was focused on advances in biology, physiology, and pathology of neoplasms affected by hormones, especially breast and prostate cancers. The stem cell function, the genetic and epigenetic interactions with hormones, the mechanisms of estrogen receptor transcription, biochemical markers and therapeutic targets in breast cancer, promotion of breast cancer carcinogenesis by progesterone, the basis for prostate cancer progression and the relevance of DNA repair processes, androgen receptor programming during prostate carcinogenesis, the metabolic stress role in tumor survival, and the diagnostic use of imaging in prostate cancer were discussed. Cancer Res; 75(7); 1–4. ©2015 AACR.

Carly Roop, RD, is a registered dietitian at the Joan Karnell Supportive Care Services (JKSCS). She provides nutrition education and support to patients while addressing nutrition-related side effects from chemotherapy and radiation. Dietitians at JKSCS provide educational nutrition programs that are open to patients as well as the community.

Move over kale, this year cauliflower is taking over menus everywhere. And the cauliflower appearing on menus at some of the top restaurants in the United States is not the same steamed cauliflower you once pushed around your plate.

Dietitians everywhere are thrilled about this comeback because cauliflower is full of cancer and disease fighting properties. Cauliflower is a cruciferous vegetable, like broccoli, Brussels sprouts, bok choy and cabbage; it belongs to the cabbage family. These sulfur-containing vegetables may have a pungent smell but they also contain phytochemicals such as Sulforaphane and Indole-3-carbinol which aid the liver in production of enzymes that block cancer-causing chemicals from damaging the body and help enhance the immune system.

This vegetable is low in calories while still being rich in vitamin C, fiber and B vitamins such as biotin, which contributes to healthy hair and nails. Cauliflower is more than crudité, there are a variety of ways one can prepare this vegetable, it can be coated with a spicy yogurt and baked tandoori style, pureed into a creamy soup, roasted with a touch of melted Parmesan, tossed in sauce for vegetarian Buffalo wings and it can even be prepared to have a couscous- like texture.

The year has just begun but I predict there is a lot more culinary fun to come with cauliflower.

Spicy Whole Roasted Cauliflower 

Serves 6 

Ingredients:

• 1 tbsp. vegetable oil 1 head of cauliflower 
• 1 ½ cups of plain Greek yogurt 
• 1 lime, zested and juiced 
• 1-2 tbsp. chili powder 
• 1 tbsp. cumin 
• 1 tbsp. garlic powder 
• 1 tsp curry powder 
• 2 tsp kosher salt 
• 1 tsp black pepper

Procedure:

1. Preheat oven to 400 °F and lightly grease a small baking sheet with vegetable oil. 
2. Trim the base of the cauliflower to remove any green leaves and the woody stem.
3. In a medium bowl, combine the yogurt with the lime zest and juice, chili powder, cumin, garlic powder, curry powder and pepper. 
4. Dunk the cauliflower in the bowl and use a brush or your hands to smear the marinade on evenly over its surface (Extra marinade can be stored in the refrigerator in an airtight container for up to 3 days). 
5. Place the cauliflower on the prepared baking sheet and roast until the surface is dry and lightly browned 35-45 minutes.
6. Let the cauliflower cool for 10 minutes before cutting it into wedges and serving alongside a big green salad

Nutrition per serving: 65 calories; < 1 gm of total fat; 7 gram of carbohydrates; 2 gm of dietary fiber; 7.5 gm of protein

March is Colorectal Cancer Awareness Month. To learn more about screening, symptoms, treatments and the latest research, we’re teaming up with 6ABC for a live, streaming web chat at 6abc.com/pennmedicine.

Colorectal Cancer – Did you know?

You may be at a higher risk for colorectal cancer as you get older, but with education and screening, you can help reduce your risk.

Screening Can Save Lives

For those 50 years and older, getting a colorectal cancer screening can be life saving.

Symptoms like the appearance of polyps in the colon or rectum, persistent stomach aches and pains, rapid weight loss and bloody bowel movements may be key indicators.

While these symptoms may be indicative of other ailments, colorectal cancer screenings can help you take the right next steps.

This March, Colorectal Cancer Awareness Month, be sure to ask the right questions, and get the facts.

Our Experts Answering Your Questions

Patients and loved ones can submit questions ahead of time and also watch the chat in full after our broadcast.

Want to know more about:

• Colorectal cancer screening?
• Risk evaluation?
• Hereditary risk?
• Cutting-edge treatment potions?
• New breakthrough clinical trials?
• Survivorship?

Please join Penn experts Gregory G. Ginsberg, MD, Timothy C. Hoops, MD and Joshua I.S. Bleier, MD on Thursday, March 19 at 4pm.