Abstract P2-11-02: Mapping Mutations: Importance of Family Geographic Origin in Portuguese High-Risk Breast/Ovarian Cancer Families for BRCA Mutation Screening

Abstract Background: Hereditary breast/ovarian cancer is mainly related with BRCA1/2 mutations. The complete screening of both genes is long and expensive due to their large number of exons and the inexistence of hotspots. Besides our most frequent BRCA mutation, c.156_157insAlu in BRCA2, the Portuguese population has many other mutations in these genes, described over the years. Aim: to evaluate the importance of screening specific mutations, found in BRCA positive families, in other probands with the same geographic origin. Patients and methods: 407 high risk breast/ovarian cancer patients were screened for c.156_157insAlu in BRCA2 (the Portuguese founder mutation), BRCA1/2 point mutations and large rearrangements. All probands were questioned about their family origin. Mutations detected in families were studied in new probands with the same geographic origin. Results: we were able to draw a map with the geographic origin of our 87 BRCA positive families. Most of our BRCA families have origin in the centre of Portugal. While searching probands with the same geographic origin as others previously detected, we found three additional mutations without screening the whole genes: 1 family from Madeira island; 1 family from the south (Algarve) and 1 non-affected individual, without previous family testing, that was positive for c.156_157insAlu in BRCA2 (her family had origin in an area of high prevalence of this mutation). Our maps also allowed the linking of several families with the same mutation and family origin, but apparently unrelated. Conclusion: Mapping geographic origin of BRCA1/2 mutations may help in the quick identification of new BRCA1/2 carriers and, in some cases, reduce the screening time. Most importantly it permits the linking of apparently unrelated families. This link permits a more accurate estimation of BRCA1/2 positive families and a better understanding of all at-risk family members. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-11-02.</jats:p

Copper(II) and Gallium(III) Complexes of <i>trans</i>-Bis(2-hydroxybenzyl) Cyclen Derivatives: Absence of a Cross-Bridge Proves Surprisingly More Favorable

Two cyclen (1,4,7,10-tetraaza­cyclododecane) derivatives bearing <i>trans</i>-bis­(2-hydroxybenzyl) arms, the 1,7-(2-hydroxybenzyl)-1,4,7,10-tetraazacyclododecane (H₂do2ph) and its cross-bridged counterpart (H₂cb-do2ph), have been synthesized, aiming toward the possible use of their copper­(II) and gallium­(III) complexes in nuclear medicine. The protonation of both compounds was studied in aqueous solution as well as their complexes with Cu²⁺ and Ga³⁺ cations. The complexes of both ligands with Ca²⁺ and Zn²⁺ metal ions were also studied due to the abundance of these cations in biological media. In mild conditions the complexes of Ca²⁺ and Ga³⁺ with H₂cb-do2ph did not form. The behavior of the two ligands and their complexes was compared by the values of the equilibrium constants, the data of varied spectroscopic techniques, the values of redox potentials of their copper­(II) complexes, and the resistance of the complexes to acid dissociation. It was expected that, as found for related pairs of cyclen and cyclam (1,4,8,11-tetraaza­cyclotetradecane) derivatives, the cross-bridged macrocyclic derivative could be an excellent ligand for the complexation of copper­(II). Additionally, the <i>N</i>-2-hydroxybenzyl groups were chosen due to their known ability to coordinate the gallium­(III) cation. Due to the small size of the latter cation and its particular propensity to form hexacoordinate complexes, it was also expected that there would be a good ability of both ligands for the uptake of Ga³⁺. Surprisingly, the results revealed that the cyclen derivative H₂do2ph is the best ligand for the coordination of Cu²⁺ and Ga³⁺ cations, not only from their thermodynamic stability as expected but also from their kinetic inertness, when compared with its cross-bridged counterpart

Copper(II) and Gallium(III) Complexes of <i>trans</i>-Bis(2-hydroxybenzyl) Cyclen Derivatives: Absence of a Cross-Bridge Proves Surprisingly More Favorable

Two cyclen (1,4,7,10-tetraaza­cyclododecane) derivatives bearing <i>trans</i>-bis­(2-hydroxybenzyl) arms, the 1,7-(2-hydroxybenzyl)-1,4,7,10-tetraazacyclododecane (H₂do2ph) and its cross-bridged counterpart (H₂cb-do2ph), have been synthesized, aiming toward the possible use of their copper­(II) and gallium­(III) complexes in nuclear medicine. The protonation of both compounds was studied in aqueous solution as well as their complexes with Cu²⁺ and Ga³⁺ cations. The complexes of both ligands with Ca²⁺ and Zn²⁺ metal ions were also studied due to the abundance of these cations in biological media. In mild conditions the complexes of Ca²⁺ and Ga³⁺ with H₂cb-do2ph did not form. The behavior of the two ligands and their complexes was compared by the values of the equilibrium constants, the data of varied spectroscopic techniques, the values of redox potentials of their copper­(II) complexes, and the resistance of the complexes to acid dissociation. It was expected that, as found for related pairs of cyclen and cyclam (1,4,8,11-tetraaza­cyclotetradecane) derivatives, the cross-bridged macrocyclic derivative could be an excellent ligand for the complexation of copper­(II). Additionally, the <i>N</i>-2-hydroxybenzyl groups were chosen due to their known ability to coordinate the gallium­(III) cation. Due to the small size of the latter cation and its particular propensity to form hexacoordinate complexes, it was also expected that there would be a good ability of both ligands for the uptake of Ga³⁺. Surprisingly, the results revealed that the cyclen derivative H₂do2ph is the best ligand for the coordination of Cu²⁺ and Ga³⁺ cations, not only from their thermodynamic stability as expected but also from their kinetic inertness, when compared with its cross-bridged counterpart

P2-13-11: Follow Up of BRCA1/2 Carriers: The Spectrum of Cancer Diagnoses in Healthy at Risk Individuals (HTR), and in Cancer Survivors (CS).

Abstract Introduction and Objective: Data from long term follow up of BRCA1/2 carriers is scarce and is mainly related to BRCA1 women. Our multidisciplinary program targets both women and men for counselling towards BRCA1/2 screening and inclusion in clinical follow up. In here we review all cancer diagnoses observed in our BRCA1/2 cohort during follow up. Methods: Review of individual records of BRCA1/2 carriers registered from January 2000 to December 2010. Follow up was calculated since BRCA1/2 post-test counselling until the last visit to the Clinic. All new cancer diagnoses and preventive surgeries were registered. Results: Two-hundred and fifty nine BRCA1/2 carriers (206 females and 53 males) were diagnosed with BRCA1/2 mutations (42 BRCA1 and 217 BRCA2). Medium follow up for all population is 25 months (1-98). At the date of initial BRCA1/2 diagnosis 99 women and 14 men were CS. Female population: Eighty-eight female CS had been previously diagnosed with breast cancer (18 bilateral cases), 18 with ovarian cancer, and 1 with biliary tract cancer. Preventive surgeries in the CS female population were: bilateral adnexectomy (33 pts) and prophylactic contralateral mastectomy (10 pts). In this CS female population, new cancers, during follow up were: Contralateral breast cancer (4 cases), peritoneal cancer (2 cases in pts with previous prophylactic surgery) and skin non-melanoma cancers (2 cases). In female HTR, 23 preventive bilateral adnexectomies and 20 bilateral mastectomies were performed. Cancer diagnoses during follow up were: breast (11), peritoneum (1 in a pt with previous prophylactic surgery), gastric (1) and M3 leukemia (1). Global failure of prophylactic adnexectomy, so far (CS+HTR): 3/56 (5%). Male population: The medium age for male CS is 73 yrs and for male HTR is 52 yrs. Male CS had mostly been previously diagnosed with BC (12; 4 bilateral) and prostate cancer (4). Other previous cancers: gastric (2), skin (2: 1Melanoma, 1 non-Melanoma), colorectal (1). One BRCA2 man with gynecomastia and prostate cancer was submitted to reduction mastectomy, as a preventive surgery. During follow up, we diagnosed second and third cancers in male CS: breast (2), prostate (6) and gastric (1). Only 1 male HTR was diagnosed with cancer: skin non-melanoma. Conclusion: The proportion of second and third cancer diagnoses in the male BRCA2 CS population is higher than in the female BRCA1/2 CS population. Small numbers and the availability of preventive surgery for women influence this observation. No data from preventive mastectomy exists for males (we have one case). The low frequency of cancer in male HTR may be due to younger age and other unknown modifier factors. Longer follow is needed. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-13-11.</jats:p

Copper(II) Complexes of Phenanthroline and Histidine Containing Ligands: Synthesis, Characterization and Evaluation of their DNA Cleavage and Cytotoxic Activity

Copper­(II) complexes have been intensely investigated in a variety of diseases and pathological conditions due to their therapeutic potential. The development of these complexes requires a good knowledge of metal coordination chemistry and ligand design to control species distribution in solution and tailor the copper­(II) centers in the right environment for the desired biological activity. Herein we present the synthesis and characterization of two ligands HL1 and H₂L2 containing a phenanthroline unit (phen) attached to the amino group of histidine (His). Their copper­(II) coordination properties were studied using potentiometry, spectroscopy techniques (UV–vis and EPR), mass spectrometry (ESI-MS) and DFT calculations. The data showed the formation of single copper complexes, [CuL1]⁺ and [CuL2], with high stability within a large pH range (from 3.0 to 9.0 for [CuL1]⁺ and from 4.5 to 10.0 for [CuL2]). In both complexes the Cu²⁺ ion is bound to the phen unit, the imidazole ring and the deprotonated amide group, and displays a distorted square pyramidal geometry as confirmed by single crystal X-ray crystallography. Interestingly, despite having similar structures, these copper complexes show different redox potentials, DNA cleavage properties and cytotoxic activity against different cancer cell lines (human ovarian (A2780), its cisplatin-resistant variant (A2780cisR) and human breast (MCF7) cancer cell lines). The [CuL2] complex has lower reduction potential (<i>E</i>_pc= −0.722 V vs −0.452 V for [CuL1]⁺) but higher biological activity. These results highlight the effect of different pendant functional groups (carboxylate vs amide), placed out of the coordination sphere, in the properties of these copper complexes

Copper(II) Complexes of Phenanthroline and Histidine Containing Ligands: Synthesis, Characterization and Evaluation of their DNA Cleavage and Cytotoxic Activity

Copper­(II) complexes have been intensely investigated in a variety of diseases and pathological conditions due to their therapeutic potential. The development of these complexes requires a good knowledge of metal coordination chemistry and ligand design to control species distribution in solution and tailor the copper­(II) centers in the right environment for the desired biological activity. Herein we present the synthesis and characterization of two ligands HL1 and H₂L2 containing a phenanthroline unit (phen) attached to the amino group of histidine (His). Their copper­(II) coordination properties were studied using potentiometry, spectroscopy techniques (UV–vis and EPR), mass spectrometry (ESI-MS) and DFT calculations. The data showed the formation of single copper complexes, [CuL1]⁺ and [CuL2], with high stability within a large pH range (from 3.0 to 9.0 for [CuL1]⁺ and from 4.5 to 10.0 for [CuL2]). In both complexes the Cu²⁺ ion is bound to the phen unit, the imidazole ring and the deprotonated amide group, and displays a distorted square pyramidal geometry as confirmed by single crystal X-ray crystallography. Interestingly, despite having similar structures, these copper complexes show different redox potentials, DNA cleavage properties and cytotoxic activity against different cancer cell lines (human ovarian (A2780), its cisplatin-resistant variant (A2780cisR) and human breast (MCF7) cancer cell lines). The [CuL2] complex has lower reduction potential (<i>E</i>_pc= −0.722 V vs −0.452 V for [CuL1]⁺) but higher biological activity. These results highlight the effect of different pendant functional groups (carboxylate vs amide), placed out of the coordination sphere, in the properties of these copper complexes

Copper(II) Complexes of Phenanthroline and Histidine Containing Ligands: Synthesis, Characterization and Evaluation of their DNA Cleavage and Cytotoxic Activity

Copper­(II) complexes have been intensely investigated in a variety of diseases and pathological conditions due to their therapeutic potential. The development of these complexes requires a good knowledge of metal coordination chemistry and ligand design to control species distribution in solution and tailor the copper­(II) centers in the right environment for the desired biological activity. Herein we present the synthesis and characterization of two ligands HL1 and H₂L2 containing a phenanthroline unit (phen) attached to the amino group of histidine (His). Their copper­(II) coordination properties were studied using potentiometry, spectroscopy techniques (UV–vis and EPR), mass spectrometry (ESI-MS) and DFT calculations. The data showed the formation of single copper complexes, [CuL1]⁺ and [CuL2], with high stability within a large pH range (from 3.0 to 9.0 for [CuL1]⁺ and from 4.5 to 10.0 for [CuL2]). In both complexes the Cu²⁺ ion is bound to the phen unit, the imidazole ring and the deprotonated amide group, and displays a distorted square pyramidal geometry as confirmed by single crystal X-ray crystallography. Interestingly, despite having similar structures, these copper complexes show different redox potentials, DNA cleavage properties and cytotoxic activity against different cancer cell lines (human ovarian (A2780), its cisplatin-resistant variant (A2780cisR) and human breast (MCF7) cancer cell lines). The [CuL2] complex has lower reduction potential (<i>E</i>_pc= −0.722 V vs −0.452 V for [CuL1]⁺) but higher biological activity. These results highlight the effect of different pendant functional groups (carboxylate vs amide), placed out of the coordination sphere, in the properties of these copper complexes

Selective Inhibition of Bruton’s Tyrosine Kinase by a Designed Covalent Ligand Leads to Potent Therapeutic Efficacy in Blood Cancers Relative to Clinically Used Inhibitors

Bruton’s tyrosine kinase (BTK) is a member of the TEC-family kinases and crucial for the proliferation and differentiation of B-cells. We evaluated the therapeutic potential of a covalent inhibitor (JS25) with nanomolar potency against BTK and with a more desirable selectivity and inhibitory profile compared to the FDA-approved BTK inhibitors ibrutinib and acalabrutinib. Structural prediction of the BTK/JS25 complex revealed sequestration of Tyr551 that leads to BTK’s inactivation. JS25 also inhibited the proliferation of myeloid and lymphoid B-cell cancer cell lines. Its therapeutic potential was further tested against ibrutinib in preclinical models of B-cell cancers. JS25 treatment induced a more pronounced cell death in a murine xenograft model of Burkitt’s lymphoma, causing a 30–40% reduction of the subcutaneous tumor and an overall reduction in the percentage of metastasis and secondary tumor formation. In a patient model of diffuse large B-cell lymphoma, the drug response of JS25 was higher than that of ibrutinib, leading to a 64% “on-target” efficacy. Finally, in zebrafish patient-derived xenografts of chronic lymphocytic leukemia, JS25 was faster and more effective in decreasing tumor burden, producing superior therapeutic effects compared to ibrutinib. We expect JS25 to become therapeutically relevant as a BTK inhibitor and to find applications in the treatment of hematological cancers and other pathologies with unmet clinical treatment

Liver injury in hospitalized patients with COVID-19: An International observational cohort study

Background: Using a large dataset, we evaluated prevalence and severity of alterations in liver enzymes in COVID-19 and association with patient-centred outcomes.MethodsWe included hospitalized patients with confirmed or suspected SARS-CoV-2 infection from the International Severe Acute Respiratory and emerging Infection Consortium (ISARIC) database. Key exposure was baseline liver enzymes (AST, ALT, bilirubin). Patients were assigned Liver Injury Classification score based on 3 components of enzymes at admission: Normal; Stage I) Liver injury: any component between 1-3x upper limit of normal (ULN); Stage II) Severe liver injury: any component &amp; GE;3x ULN. Outcomes were hospital mortality, utilization of selected resources, complications, and durations of hospital and ICU stay. Analyses used logistic regression with associations expressed as adjusted odds ratios (OR) with 95% confidence intervals (CI).ResultsOf 17,531 included patients, 46.2% (8099) and 8.2% (1430) of patients had stage 1 and 2 liver injury respectively. Compared to normal, stages 1 and 2 were associated with higher odds of mortality (OR 1.53 [1.37-1.71]; OR 2.50 [2.10-2.96]), ICU admission (OR 1.63 [1.48-1.79]; OR 1.90 [1.62-2.23]), and invasive mechanical ventilation (OR 1.43 [1.27-1.70]; OR 1.95 (1.55-2.45). Stages 1 and 2 were also associated with higher odds of developing sepsis (OR 1.38 [1.27-1.50]; OR 1.46 [1.25-1.70]), acute kidney injury (OR 1.13 [1.00-1.27]; OR 1.59 [1.32-1.91]), and acute respiratory distress syndrome (OR 1.38 [1.22-1.55]; OR 1.80 [1.49-2.17]).ConclusionsLiver enzyme abnormalities are common among COVID-19 patients and associated with worse outcomes

At-admission prediction of mortality and pulmonary embolism in an international cohort of hospitalised patients with COVID-19 using statistical and machine learning methods

By September 2022, more than 600 million cases of SARS-CoV-2 infection have been reported globally, resulting in over 6.5 million deaths. COVID-19 mortality risk estimators are often, however, developed with small unrepresentative samples and with methodological limitations. It is highly important to develop predictive tools for pulmonary embolism (PE) in COVID-19 patients as one of the most severe preventable complications of COVID-19. Early recognition can help provide life-saving targeted anti-coagulation therapy right at admission. Using a dataset of more than 800,000 COVID-19 patients from an international cohort, we propose a cost-sensitive gradient-boosted machine learning model that predicts occurrence of PE and death at admission. Logistic regression, Cox proportional hazards models, and Shapley values were used to identify key predictors for PE and death. Our prediction model had a test AUROC of 75.9% and 74.2%, and sensitivities of 67.5% and 72.7% for PE and all-cause mortality respectively on a highly diverse and held-out test set. The PE prediction model was also evaluated on patients in UK and Spain separately with test results of 74.5% AUROC, 63.5% sensitivity and 78.9% AUROC, 95.7% sensitivity. Age, sex, region of admission, comorbidities (chronic cardiac and pulmonary disease, dementia, diabetes, hypertension, cancer, obesity, smoking), and symptoms (any, confusion, chest pain, fatigue, headache, fever, muscle or joint pain, shortness of breath) were the most important clinical predictors at admission. Age, overall presence of symptoms, shortness of breath, and hypertension were found to be key predictors for PE using our extreme gradient boosted model. This analysis based on the, until now, largest global dataset for this set of problems can inform hospital prioritisation policy and guide long term clinical research and decision-making for COVID-19 patients globally. Our machine learning model developed from an international cohort can serve to better regulate hospital risk prioritisation of at-risk patients