looking back at 20 years at UIC-- the Dharma of Science

I thought I would start with a confession—I have my PhD in Biochemistry and I have actually never taken a course in Physiology. One of my mottoes has been “pose as an expert and soon become one” and I used this to my advantage in 1989 when I was hired in my first professional position, Assistant Professor at UIC. I was recruited from Michigan in the guise of a reproductive physiologist because of the work I’d done in Anita Payne’s lab in steroid hormone biochemistry. If I had to describe myself then, I would have said I was a steroid biochemist. The physiology of gonadal steroidogenesis is reproductive endocrinology. And so, I morphed into my new role as a reproductive biologist. I spent 8 hours per lecture that first year preparing to teach in the Pharmacy course about the menstrual cycle and I stayed just enough ahead of the material to make it through the first year—teaching all of endocrine physiology in the Allied Health course. I continued to give the same lectures for 18 years until PHYB341 was replaced by the new GEMS Physiology course.

Through that time I became an endocrinologist, a reproductive biologist, and a physiologist. All of our work was in vitro in primary cultures of Leydig cells, a craft I’d helped to develop and perfect in Michigan. Very few labs had the expertise in Leydig cells or molecular biology that my lab had, and this served us well. Based on the observation that Leydig cells and testicular interstitial macrophages are closely associated, I postulated that since these cells were physically associated, they must be functionally associated, and proposed that macrophage elaborated factors must affect Leydig cell function and this hypothesis guided the next 15 years of work in my lab. We identified a new role for pro-inflammatory cytokines, as regulators of steroidogenesis, and helped to foster a burgeoning interest in immune-endocrine interactions. But the question which emerged as I taught my lectures in endocrine physiology, was does what we see in vitro happen in vivo? And so, I embarked on a series of studies in systems physiology. To activate macrophages in vivo we used LPS, bacterial endotoxin derived from the cell walls of gram negative bacteria, reasoning that LPS would activate the immune system without having to make the animals septic with bacteria. We showed in vivo what happened in vitro—only more so (to paraphrase Jacob Manot what is true for elephants is true for bacteria, only more so). We observed what I still believe is an astounding effect. Within minutes, literally, after injecting a single sub-letal dose of LPS into mice, serum testosterone levels begin to fall. It becomes significant after 15 minutes and at 2 hours serum testosterone is reduced by over 85%! This is a highly reproducible finding which we observed in 100s of mice. The problem was we couldn’t reproduce this in vitro. We determined that the decrease is due to the inhibition of StAR. Karen Hales expressed the full length StAR protein and bacteria and made a superb antibody which works in all species from teleost to human and spawned many fruitful collaborations for us. It made sense that shutting off steroid biosynthesis by preventing the substrate cholesterol to get into the mitochondria where the first enzymatic step in steroid hormone synthesis takes place would account for the rapid shut off. But how did LPS cause this? We could not cause this to happen in vitro with cytokines and were stumped for quite some time. And then our friend Jossey Orly came to visit from Israel and using our antibody he demonstrated that if you treat Leydig cells in vitro with CCCP, a protonphore that disrupts the mitochondria, you can replicate the in vivo effect. We quickly repeated this and realized that LPS somehow causes Leydig cell mitochondria perturbation! Now we could model this in vitro and explored the role of the mitochondria itself in the process of steroidogenesis. We still did not know what mediated the effect in vivo, but soon determined that it was reactive oxygen elaborated by the testicular macrophages that affected the adjacent Leydig cells. I was fortunate to have John Allen and Thorsten Diemer in my lab at the time and we published a series of seminal papers which changed the way people think about steroid hormone production—that it is the mitochondria which is the key regulator, not StAR or the enzymes; and established the importance of oxidative stress and inflammation as key mediators of immune-endocrine interactions.

Despite the number of highly cited papers in high impact journals coming from these studies, the NIH did not deem this line of work fundable, .and I knew I was going to have to do something other than study immune-endocrine regulation of Leydig cell steroidogenesis. One of the reviewers said “Dr. Hales has found another bad thing that LPS does to Leydig cells,” dismissing the whole body of work which demonstrated the role of oxidative stress and the mitochondria in regulating steroidogenesis. So, I tried alcohol vs. Leydig cells, diabetes vs. Leydig cells, Dioxin vs. Leydig cells and aging vs. Leydig cells—which all have as a central theme perturbation of mitochondrial function via cumulative oxidative insult. 5 or 6 more grants attempts later, I was still unfunded and facing extinction. This became a real test of faith and I seriously pondered changing professions, but to what, I was not sure. I had been on the same path for my whole career.

Soon after the birth of my son I found my way back to the path. Desperation breeds contemplation and sitting into the late hours rocking baby Ryder gave me lots of time to think. I had envisioned years of studies which explored the mechanisms through which reactive oxygen species controlled basic biological processes—mitochondrial energetics, gene expression, serving as signaling molecules as well as electrophiles. My grandiose visions however, did not get me funded. The sentiment in the NICHD for funding the male was characterized by a cynicism- statement s coming from NIH staffers like “you can cut somebody’s balls off and they won’t die, so why study them;” and “since we have Viagra and ICSI we don’t need to study the male;” and “honestly, Buck, outside of Andrology no one really cares about Leydig cells.” So much for being one of the world experts on Leydig cells, a lot of good that was doing me. Indeed, a true test of faith. Racked by insomnia I started reading books about how to sleep. They offered practical advice about coffee consumption late in the day, and regular habits, etc. but one thing each of these texts had in common was the suggestion of meditation. I was in Borders one evening and I picked up a book by Rob Nairn entitled “What is Meditation?” which turned out to be a primmer in Buddhism. The book really spoke to me, I realized I’d been searching for this path for my entire adult life and had oft been exposed to “eastern “ philosophy during my years in Boulder—and beyond. For years I had been practicing yoga and was ever more interested in the esoteric aspects of the practice and the interaction between Yoga and Buddhism. I began to read more and more books about Buddhism and discovered the Buddha within. The power of mediation and the tenets of The Dharma were concepts so familiar to me that I felt like I’d found my way. The Way. Buddhism is not a religion as much as it is a process and I had a new tool in my psychological toolkit to help me find my way back to the path. My own understanding of Dharma was well stated in the January 19, 2007 edition of "dailyIinsight" from Yoga Journal:

• "Your personal Dharma is the path you follow toward the highest expression of your own nature and toward the fulfillment of your responsibilities to yourself, to others, to your society, and to the planet. In the Bhagavad Gita, Krishna often speaks of dharma as something inborn, a life calling that each of us has been given and from which we depart at our peril. But he also uses the word to mean right action, and for most of us, personal dharma comes down to that most basic question: What is the right thing for me to do now? Or, Given my nature, my skills, and my personal preferences, what actions should I take to support the greater good?"

What actions could I take to support The Greater Good? What path do I follow toward fulfillment? I kept the faith, I believed in what I do and what I am and was lucky to be nimble enough to adapt my research efforts into a more sustainable effort.

If you were to ask me now how I define myself, I would say that I am a cancer biologist. How I went from steroid biochemist to cancer biologist is not as much of a leap as it might sound. Reasoning that in response to immune insult, potent oxidants and inflammatory mediators combine to perturb, and shut down hormone production—but in steroid target tissues, oxidative stress and inflammation work in concert with hormones and the result in carcinogenesis. So instead of looking at the testis, which evidently no one cares about, I looked at the prostate, which a lot of people care about. With Gail Prins help we looked at oxidative stress in a rat model of prostate cancer, and I was able to get funded by the local American Cancer Society who have a grant mechanism to attract new investigators into cancer research. That one year of ACS funding was what helped me turn the corner. I never was able to get NIH funding for the prostate cancer project, but one day, I heard a Gyn Onc Jean Hurteau here at UIC say that ox stress and inflammation are thought to be important to the pathogenesis of OvCa which caught my attention. He then said that the only good model for OvCa is the chicken. I immediately thought of my old friend, with whom I collaborated since I was in Anita’s lab, Janice Bahr, who is a world expert on the reproductive biology of chickens. All of our antibodies worked in the chicken so I had followed her work and decided to see if she would like to collaborate. We were successful in getting a pilot grant from the DOD testing the inflammation hypothesis in the hen model and this funding really got me going again. One of Hurteau’s interests was in omega-3 therapy for OvCa. So I was keen on the literature but couldn’t imagine gavaging hundreds of chickens every day with fish oil. Then I was in Whole Foods one day and noticed the omega eggs in the cooler and wondered how they were able to get omega into chicken eggs. Well they feed them flaxseed, the richest vegetable source of omega-3 fatty acids. I proposed using the hen model as a pre-clinical test for chemoprevention of OvCa with flaxseed and got funded by the AICR, and then the NCCAM. Using the hen as a model for dietary intervention to prevent or treat cancer turned out to be a very tractable idea, and we have also been funded by the NCI to determine if broccoli can prevent OvCa in hens. And yes, chickens do eat broccoli.

These functional foods work by interacting with specific molecular targets do decrease oxidative stress and inflammation. We are able to draw upon the expertise we developed looking at these forces in Leydig cells and have expanded this to the study of a very important and deadly disease. I remain steadfast in my belief that if you do good science and let biology lead the way, you will answer important questions, whose answers raise more questions which guide you further down the path. It is true that you have to be nimble as you find your way along the path and be able to surmount the obstacles you encounter along the way. My Dharma is science and biology guides the way. All of the work we did in Leydig cells is still important and has helped create significant interest in the role of immune and endocrine interactions in the maintenance of homeostasis, as well as mediating pathologies. But our current work in ovarian cancer is more important because it holds the promise of discovering early detection for ovarian cancer, prevention and treatment of the disease, and most important, it provides hope for women who have the disease, hope that their daughters, sisters and friends may not have to suffer what they are dying from.

There is a lot of public interest in cancer biology, antioxidants, dietary intervtion, natural, and alternative medicine-- so I have decided to launch a new blog which I will call CancerChix, so that I can share my thoughts and observations on these topics without diluting these posts with my rambling narratives about bicycle commuting, diet and exercise and politics.