Edward J. Keenan

p95HER-2 Predicts Worse Outcome in Patients with HER-2-Positive Breast Cancer

Background: The HER-2 receptor undergoes a proteolytic cleavage generating an NH2-terminally truncated fragment, p95HER-2, that is membrane-associated and tyrosine-phosphorylated. We have reported that p95HER-2, but not the full-length receptor, p185HER-2, correlated with the extent of lymph node involvement in patients with breast cancer and its expression was significantly enhanced in nodal metastatic tissue. These facts suggested an important role for p95HER-2 either as a marker or cause of metastasis and poor outcome in breast cancer. In this work, we have studied the prognostic value of p95HER-2 in breast cancer. Methods: Primary breast tumor tissues (n = 483) were from surgical resections conducted in hospitals in two different countries: the U.S. (n = 334) and Spain (n = 149). HER-2 protein forms, including p185HER-2 and p95HER-2, were examined in extracts of primary breast tumors by Western blot analysis. The levels of the two forms (high or low) were tested for association with other clinicopathologic factors and for correlation with disease-free survival. Results: The median follow-up was 46 months. A high level of p95HER-2 in primary tumor tissue correlated with reduced 5-year disease-free survival (hazard ratio, 2.55; 95% confidence interval, 2.13-8.01; P < 0.0001). The median time for disease-free survival was 32 versus 139 months in patients with low levels of p95HER-2. In comparison, high levels of the full-length p185HER-2 did not significantly correlate with poor outcome (P > 0.1). Multivariate analysis revealed that high p95HER-2 was an independent predictor of disease-free survival (hazard ratio, 1.59; 95% confidence interval, 1.246-1.990; P = 0.0004). Conclusions: p95HER-2 expression is an independent prognostic factor in breast cancer and defines a group of patients with HER-2-positive breast cancer with significantly worse outcome.

Marked Decline in Trabecular Bone Mineral Content in Healthy Men with Age: Lack of Association with Sex Steroid Levels

To define the association of age‐related changes in bone mineral content to gonadal function in normal men, we measured radial (largely cortical) and vertebral (largely trabecular) bone mineral content (BMC), testosterone (total and free), estrone and estradiol‐17B levels in 62 healthy subjects, ages 30 to 92. Radial BMC fell 2 to 3.4% per decade but vertebral trabecular BMC declined more rapidly at 12% per decade. Of the sex steroids measured the only statistically significant change occurred in free testosterone levels which decreased with age (r = −.57, P < .0001). Free testosterone levels correlated significantly with trabecular vertebral BMC (r = .458, P < .0002) but not with bone mineral measures at the predominantly cortical radial sites. However, by multiple regression analysis free testosterone did not add to the effect of age on vertebral BMC. There were no associations of total testosterone, estrone, or estradiol levels to bone mineral content at any of the three sites measured in these healthy men. Age‐related declines in male gonadal function do not appear to be of primary importance in male age‐related bone loss.

Somatostatin analogs: angiogenesis inhibitors with novel mechanisms of action

Eugene A. Woltering1, James C. Watson1, R. Christian Alperin-Lea1, Chakravarthi Sharma1, Edward Keenan2, Daryl Kurozawa2 and Rosemary Barrie2 1Louisiana State University School of Medicine, Department of Surgery, Section of Surgical Endocrinology, 1542 Tulane Avenue, New Orleans, Louisiana 70122 and Veterans Administration Medical Center – New Orleans, 1601 Perdido Street, New Orleans, Louisiana 70146; 2Oregon Health Science University, Department of Surgery, Division of Surgical Oncology, 3181 Sam Jackson Park Road, Portland, Oregon 98702, USA

Effect of clomiphene citrate treatment on endometrial estrogen and progesterone receptor induction in women

A direct adverse effect of clomiphene citrate on the endometrium has been presumed, and interference with estrogen receptor-mediated endometrial estrogen receptor and progesterone receptor induction has been implicated as the mechanism responsible for an increased incidence of luteal phase deficiency in association with clomiphene citrate treatment. To clarify the net influence of clomiphene administration on endometrial steroid receptor induction, we studied five normal ovulatory women, in both a spontaneous and clomiphene-induced (150 mg/day, cycle days 5 to 9) ovulatory cycle. From cycle day 11 blood samples were obtained daily and urinary luteinizing hormone determinations were performed twice daily. Endometrial biopsy was performed on the day of the urinary luteinizing hormone surge and again 13 days after the surge. Serum levels of follicle-stimulating hormone and luteinizing hormone were determined by immunoradiometric assay, estradiol and progesterone by radioimmunoassay, and clomiphene citrate isomer concentrations in treatment cycles by reversed-phase high-performance liquid chromatography and fluorescence detection. Total, cytosolic, and salt-extracted nuclear endometrial estrogen receptor and progesterone receptor concentrations were determined by enzyme-linked immunoassay. Serum estradiol was threefold to fivefold higher (p less than 0.05) in clomiphene-induced than in spontaneous cycles 8 and 10 days before the luteinizing hormone surge, and progesterone was increased (p less than 0.05) from the day of the surge to end of the cycle. Serum enclomiphene rose to plateau between 12 and 6 days before the luteinizing hormone surge (4.1 +/- 0.8 ng/ml, mean +/- SE, n = 19) and fell thereafter to less than 1.0 ng/ml. Zuclomiphene levels increased rapidly between 14 and 8 days before the surge (53.9 +/- 2.8 ng/ml, mean +/- SE, n = 5) and then decreased gradually but remained elevated throughout the luteal phase (29.0 +/- 1.2 ng/ml, mean +/- SE, n = 33). Late luteal endometrial histology was abnormal in one of four available treatment cycle specimens, but the endocrine characteristics and number and subcellular distribution of estrogen receptor and progesterone receptor in the abnormal cycle were not different from those of normal, in-phase cycles.(ABSTRACT TRUNCATED AT 250 WORDS)

α2-Adrenergic receptor levels in obstructive and spastic Raynaud's syndrome

Abstract The present study examines the hypothesis that alterations in the activity of α 2 -adrenergic receptors (A 2 R) may underlie the clinical vasospasm seen in patients with Raynauds syndrome. Platelets were isolated from 13 normal subjects, from 50 patients with vasospastic Raynauds syndrome, and from 20 patients with obstructive Raynauds syndrome and A 2 R levels measured. Binding capacity as determined in femtomoles per milligram of protein (fmol/mg of protein) and affinity were measured by Scatchard plot analysis. In a separate experiment normal human platelets were incubated with either buffer, normal serum, or serum from patients with spastic Raynauds syndrome and A 2 R levels were measured. A 2 R levels in normal subjects averaged 112 ± 18 fmol/mg; in the patients with spastic Raynauds syndrome, 191 ± 14 fmol/mg, p 0.05 (ns). Of the patients with spastic Raynauds syndrome, 26% had values that were less than the mean value of the normal subjects (69 ± 7 fmol/mg, p 2 R levels decreased after incubation with serum from patients who had spastic Raynauds syndrome by 17.4 ± 3.1 fmol/mg (p 2 R levels, which may constitute a primary pathophysiologic abnormality underlying this condition. The presence of subnormal A 2 R levels in a portion of the patients and the finding of a decrease in measurable A 2 R levels after incubation in serum from patients with spastic Raynauds syndrome suggests the possibility of receptor modulation as a mechanism for increased cellular receptor synthesis. (J VASC SURG 1987;5:38-45.)

Multiple simultaneous and sequential estrogen receptor values in patients with breast cancer.

Fifty-nine women had multiple estrogen receptor assays done, either simultaneously or sequentially. Eighty-six percent of the patients who had multiple synchronous estrogen receptor assays from various metastatic sites showed no significant discrepancy in estrogen receptor values. When estrogen receptor assays were done sequentially without intervening therapy, 83.5 percent of the patients maintained their initial positivity or negativity. However, when the second estrogen receptor determination was preceded by either chemotherapy or hormonal therapy, 33 percent of the patients had a significant discrepancy in estrogen receptor values. The most common discrepancy was estrogen receptor-positive tumors becoming estrogen receptor-negative, although a small number of patients were found whose receptor values became more positive after hormonal ablation.

Principles of endocrine pharmacology

1. Introduction and General Mechanisms of Hormonal Actions.- 1.1. History and Scope of Endocrine Pharmacology.- 1.2. General Concepts of Hormone Actions.- 1.2.1. Hormone Receptor/Acceptors.- 1.2.2. Hormone Substitutes.- 1.2.3. Hormone Antagonists.- 1.2.4. Hormone Synthesis Inhibitors.- 1.3. Hormonal Feedback Systems.- Recommended Readings.- 2. Pharmacology of Adenohypophyseal Hormones.- 2.1. Factors Modifying Adenohypophyseal Secretion: Hypophysiotrophic Hormones.- 2.1.1. CRH.- 2.1.2. TRH.- 2.1.3. LH/FSH-RH (GnRH).- 2.1.4. GH-RF and GH-IF (SRIF).- 2.1.5. PRF and PIF.- 2.1.6. MRF and MIF.- 2.2. Pharmacology of Anterior Pituitary Hormones.- 2.2.1. TSH.- 2.2.2. STH or GH.- 2.2.3. ACTH.- 2.2.4. FSH/LH.- 2.2.5. PRL.- Recommended Readings.- 3. Posterior Pituitary Hormones, Oxytocics, and Prostaglandins.- 3.1. Posterior Pituitary Hormones.- 3.1.1. History.- 3.1.2. Synthesis, Transport, and Release.- 3.1.3. Antidiuretic Hormone.- 3.1.4. Oxytocin.- 3.2. Ergot Alkaloids.- 3.2.1. Chemistry.- 3.2.2. Mechanism of Action and Biochemical Effects.- 3.2.3. Therapeutic Uses and Preparations.- 3.2.4. Adverse Effects.- 3.3. Prostaglandins: Reproductive Actions.- 3.3.1. Chemistry.- 3.3.2. Mechanism of Action and Biochemical Effects.- 3.3.3. Therapeutic Uses and Preparations.- 3.3.4. Adverse Effects.- 3.3.5. Prostaglandin Inhibitors.- 3.3.6. Uterine Relaxants.- Recommended Readings.- 4. Thyroid and Antithyroidal Drugs.- 4.1. Thyroid.- 4.1.1. History.- 4.1.2. Central Regulation of the Thyroid.- 4.1.3. Chemistry, Biosynthesis, Secretion, and Metabolism.- 4.1.4. Thyroid Hormone Receptors.- 4.1.5. Biochemical Actions.- 4.1.6. Management of Hypothyroidal States.- 4.1.7. Therapeutic Uses and Preparations.- 4.1.8. Adverse Effects.- 4.2. Antithyroidal Agents.- 4.2.1. Thyrotoxicosis (Hyperthyroidism).- 4.2.2. Drugs Used in the Management of Thyrotoxicosis.- 4.3. Drug Interactions and Thyroid Function Tests.- Recommended Readings.- 5. Parathyroid Hormone and Calcitonin.- 5.1. Introduction.- 5.2. History.- 5.3. Parathyroid Hormone.- 5.3.1. Chemistry.- 5.3.2. Biosynthesis, Secretion, and Metabolism.- 5.3.3. Mechanism of Action.- 5.3.4. Physiological and Pharmacological Actions.- 5.3.5. Preparations.- 5.3.6. Therapeutic Uses.- 5.3.7. Adverse Effects.- 5.4. Calcitonin.- 5.4.1. Chemistry.- 5.4.2. Biosynthesis, Secretion, and Metabolism.- 5.4.3. Mechanism of Action.- 5.4.4. Physiological and Pharmacological Actions.- 5.4.5. Preparations.- 5.4.6. Therapeutic Uses.- 5.4.7. Adverse Effects.- Recommended Readings.- 6. Androgenic and Anabolic Steroids.- 6.1. Introduction.- 6.2. History.- 6.3. Chemistry.- 6.4. Biosynthesis, Secretion, and Metabolism.- 6.5. Mechanism of Action.- 6.6. Physiological and Pharmacological Actions of Androgens.- 6.7. Preparations.- 6.8. Therapeutic Uses.- 6.9. Adverse Effects.- 6.10. Weak or Impeded Androgens.- 6.11. Androgen Antagonists.- Recommended Readings.- 7. Estrogens and Antiestrogenic Drugs.- 7.1. Estrogens.- 7.1.1. Introduction.- 7.1.2. History.- 7.1.3. Chemistry.- 7.1.4. Biosynthesis, Secretion, and Metabolism.- 7.1.5. Mechanism of Action.- 7.1.6. Physiological Actions.- 7.1.7. Therapeutic Uses.- 7.1.8. Preparations.- 7.1.9. Adverse Effects.- 7.2. Antiestrogens.- 7.2.1. Clomiphene.- 7.2.2. Tamoxifen.- Recommended Readings.- 8. Progestins and Oral Contraceptives.- 8.1. Progestins.- 8.1.1. Introduction.- 8.1.2. History.- 8.1.3. Chemistry.- 8.1.4. Biosynthesis, Secretion, and Metabolism.- 8.1.5. Mechanism of Action.- 8.1.6. Physiological Effects.- 8.1.7. Therapeutic Uses.- 8.1.8. Preparations.- 8.1.9. Adverse Effects.- 8.2. Oral Contraceptives.- 8.2.1. Preparations.- 8.2.2. Mechanism of Action.- 8.2.3. Adverse Effects.- Recommended Readings.- 9. Adrenocorticosteroid Drugs.- 9.1. Introduction.- 9.2. History.- 9.3. Chemistry.- 9.4. Biosynthesis, Secretion, and Metabolism.- 9.5. Mechanism of Action.- 9.6. Physiological and Pharmacological Actions.- 9.7. Preparations.- 9.8. Therapeutic Uses.- 9.9. Adverse Effects.- 9.10. Inhibitors of Adrenocortical Steroid Biosynthesis.- Recommended Readings.- 10. Insulin and Oral Hypoglycemic Agents.- 10.1. Insulin.- 10.1.1. History of Diabetes Mellitus.- 10.1.2. Chemistry of Insulin.- 10.1.3. Secretion and Metabolism.- 10.1.4. General Biochemical Events and Actions.- 10.1.5. Insulin Receptors.- 10.1.6. Receptor-Mediated Internalization.- 10.1.7. Factors Affecting Insulin and Insulin Resistance.- 10.1.8. Uses and Preparations.- 10.1.9. Adverse Effects.- 10.2. Oral Hypoglycemic Agents.- 10.2.1. Sulfonylureas.- 10.2.2. Biguanides.- 10.3. Glucagon.- 10.4. Somatostatin.- 10.5. Nonhormonal Hyperglycemic Agents.- Recommended Readings.- 11. Effects of Drugs on the Endocrine System.- 11.1. Introduction.- 11.2. Basic Mechanisms of Drug-Hormone Interactions.- 11.3. Effects of Drugs on Adenohypophyseal Function.- 11.4. Effects of Drugs on Neurohypophyseal Function.- 11.5. Effects of Drugs on Lactation and Their Presence in Milk.- 11.6. Effects of Drugs on Hormone Transport.- 11.7. Effects of Drugs on Steroidogenesis.- 11.8. Effects of Drugs on Gonadal Function.- 11.9. Effects of Drugs on Pancreatic Function.- 11.10. Effects of Drugs on Thyroid Function.- 11.11. Effects of Drugs on Laboratory Analyses.- Recommended Readings.

Insulin and Oral Hypoglycemic Agents

About 100 years have elapsed since the classic experiments of von Mering and Minkowski demonstrated that pancreatomized dogs exhibited signs and symptoms resembling those seen in diabetes mellitus. The pioneering efforts of Banting and Best revealed that pancreatic extracts could sustain the life of patients suffering from severe diabetes, thereby providing the link between insulin deficiency and the disease. Insulin was subsequently crystallized by Abel and was eventually chemically synthesized in the laboratory. Recently, synthetic insulin derived from recombinant DNA technologies has been approved for clinical trials. Therapy employing animal insulin has been used in the clinical management of diabetes mellitus for many years. Despite the experience with hormone replacement therapies, it is now recognized that diabetes mellitus is a very complex metabolic disorder, and the simple concept that its pathogenesis is due solely to insulin deficiency is no longer tenable. Indeed, contributing to the reduced production of insulin are contributing factors such as excess glucagon, which aggravate both hyperglycemia and ketosis. Insulin resistance, as demonstrated in insulin-dependent diabetes mellitus (IDDM) (i. e., type I), is yet another complicating factor and may be due to both a decrease in insulin receptors and a postreceptor defect.

Endocrine ablation for metastatic breast cancer: A reappraisal of hormone receptors☆

Other indicators of hormone sensitivity, besides estrogen receptor (ER) content, such as response to oophorectomy, antiestrogens, prolactin suppression, and correlation with progesterone receptors (PR), were evaluated in the hope of further improving selectivity and response of patients undergoing endocrine ablation for metastatic breast cancers. 225 patients have undergone full endocrine ablation in the last 30 years at this hospital, 208 by adrenalectomy and 17 by hypophysectomy. 206 of these patients could be retrospectively reviewed, and of these there were objective responses to therapy in 50% of patients. ER analyses were performed in 1 or more breast cancer specimens in 113 of these patients. ER study showed that a patient who was ER+ and responded to a functional test of endocrine sensitivity had a 70-80% chance of also benefiting from adrenalectomy or hypophysectomy. Conversely, patients with absent or unknown hormone receptors who failed therapeutic trials of endocrine sensitivity had little or no chance of responding to major ablation; these cases are best treated with multiagent chemotherapy. The value of sequentially treating selected patients with endocrine manipulation in addition to chemotherapy was also studied. Patients who failed to respond to endocrine manupulation survived slightly over 2 years on chemotherapy, whereas patients who responded to major ablation lived with metastases an average of 4 years, whereas complete responders lived with metastatic disease an average of 6 years. By life table analysis, total survival of ER+ vs. ER- patients as well as responders vs. nonresponders was highly significant

HER-2 gene amplification in human breast cancer without concurrent HER-2 over-expression

BackgroundTesting for human epidermal growth factor receptor-2 (HER-2) in breast cancer is performed by either immunohistochemistry (IHC) or in situ hybridization (ISH). The growth factor receptor-bound protein-7 (GRB7) gene is in close proximity to HER-2 on chromosome 17q11-12 and codes a signal transduction molecule shown to be an independent adverse marker in breast cancer.MethodsHER-2 and GRB7 protein expression from 613 frozen breast tumors was determined by Western analysis. HER-2 protein results were confirmed with IHC. Commercial HER-2 FISH was performed on a subset of tumors with multi-probe FISH used to assess the extent of HER-2 gene amplification. mRNA expression was determined by Multi-plex RT-PCR.ResultsSeven tumors with GRB7 protein over-expression scored HER-2 FISH amplified but had no HER-2 protein over-expression. Four of the 7 tumors showed elevated GRB7 but not HER-2 mRNA over-expression. The breast cancer cell line HCC3153 did not over-express HER-2 protein but showed HER-2 FISH amplification of a limited segment around the HER-2 gene. Ten breast cancer tumors from the TCGA database had gene copy number increases around HER-2 without HER-2 mRNA or protein over-expression.ConclusionsA subset of human breast cancers that test positive with FISH for HER-2 gene amplification do not over-express HER-2 protein. One mechanism for this discordance is the incomplete amplification of the smallest HER-2 region of chromosome 17q11-12, which includes GRB7. HER-2 gene amplification without protein over-expression is clinically significant because patients with such tumors are unlikely to benefit from HER-2 targeted therapy.