Mouse organoids as an in vitro tool to study the in vivo intestinal response to cytotoxicants.

Cross-species comparison of drug responses at the organoid level could help to determine the human relevance of findings from animal studies. To this end, we first need to evaluate the in vitro to in vivo translatability of preclinical organoids. Here, we used 5-fluorouracil (5-FU) as an exemplar drug to test whether the in vivo gut response to this cytotoxicant was preserved in murine intestinal organoids. Mice treated with 5-FU at 20 or 50 mg/kg IV (low and high dose, respectively) displayed diarrhea at clinically relevant exposures. 5-FU also induced intestinal lesions, increased epithelial apoptosis, and decreased proliferation in a dose-dependent manner. To enable comparison between the in vitro and in vivo response, top nominal in vitro drug concentrations that caused significant cytotoxicity were chosen (dose range 1-1000 µM). The inferred intracellular concentration in organoids at 1000 µM was within the tissue exposure range related to intestinal toxicity in vivo. 5-FU at ≥ 100 µM decreased ATP levels and increased Caspase-3 activity in intestinal organoids. In keeping with the in vivo findings, 5-FU increased the percentage of Caspase-3-positive cells and reduced Ki67 staining. At the transcriptome level, there was an overlap in the activity of pathways related to 5-FU's mode of action, lipid and cholesterol metabolism and integrin signaling across in vivo gut and organoids. The predicted activity state of upstream regulators was generally well preserved between setups. Collectively, our results suggest that despite their inherent limitations, organoids represent an adequate tool to explore the intestinal response to cytotoxicants.

Single-agent estramustine phosphate (EMP) is active in advanced breast cancer after failure with anthracyclines and taxanes.

PURPOSE: Estramustine phosphate (EMP) is an oral cytotoxic agent that depolymerizes tubuline, a mechanism of action that has been revisited during the last decade. Because of its lack of haematological toxicity and favourable tolerance profile, EMP is a good candidate for palliative chemotherapy. The aim of the study was to assess its tolerance and efficacy in advanced breast cancer after failure with usual regimens. PATIENTS AND METHODS: Patients with a life expectancy of at least 12 weeks and bi-dimensionally measurable disease having received at least 1 line of chemotherapy (including taxanes and/or anthracyclines) for advanced breast cancer (ABC) were eligible. EMP was given daily at a dose of 10 mg/kg until disease progression, unacceptable toxicity or patient refusal to continue chemotherapy. RESULTS: Forty patients were included between June 1998 and December 1999. Patients had previously received one to eight chemotherapy regimens (median is two) for ABC. Twenty-two patients (55%) had visceral involvement and eighteen patients (45%) had osseous, chest wall or soft tissue metastases. Adverse events leading to early interruption of EMP were grade 2 allergy (n = 1), grade 2-3 nausea (n = 6), deep-vein thrombosis (n = 1), grade 3 sepsis (n = 1). One patient died at twenty-four weeks from pulmonary embolism, and another at fourteen weeks from unknown cause. Seven objective responses were observed (17.5%; 95% confidence interval (CI): 6%-30%). Median time to failure was 24 weeks (14-52+) in responding patients. All objective responses but one were observed in patients with visceral metastases. In 10 other patients (25%), disease remained stable with a median time to failure of 27 weeks (16-50); 6 of these experienced a decrease of consumption of analgesics or an improvement of performance status. CONCLUSION: EMP is an active drug in ABC after failure with taxanes and anthracyclines, whose tolerance profile appears favourable.

[Phase I trial of a recombinant human interleukin 2. Results in patients with disseminated solid tumors]. / Essai de phase I d'une interleukine 2 humaine recombinante. Résultats chez les malades ayant une tumeur solide disséminée.

A phase I trial of Roussel-Uclaf recombinant human interleukin 2 (IL 2) was performed on 31 cancer bearing patients of the Institut Gustave-Roussy, Villejuif, and the Institut Curie, Paris. This study allowed to define a schedule for administration of IL 2 in continuous infusion over 5 day cycles. This schedule is manageable in patients without major visceral failure. It is reproducibly feasible in conventional medical oncology units, without specialized intensive care facilities. Toxicities, although numerous, are acceptable for IL 2 doses below 24,000,000 IU/m2/day. There is a close relationship between secondary effect severity and IL 2 doses received. Main toxicities were: fever with chills, fatigue and general discomfort in 23 patients, nauseas and vomiting in 12, diarrhea in 10 and cutaneous rashes with erythema and dermal vascularitis in 13. One peculiar feature of this study was the minimal occurrence of manifestation related to leaky capillary syndrome prominant in other studies. Oliguria, functional renal failure and edema were observed in only 4 patients with functionally unique kidney. Five patients had severe anemia, 2 grade III thrombocytopenia, 1 grade IV hepatic cytolysis, 4 severe confusion episodes and 2 hypothyroidism with anti-thyroid microsome auto-antibodies. All these toxicities were reversible after withdrawal of IL 2 treatment. During this phase I trial, 3 therapeutic objective responses were observed, all 3 occurring in patients with metastatic melanoma treated with IL 2 doses equal to, or above 16,000,00 IU/m2/d. Recombinant IL 2 Roussel-Uclaf thus can be administered through a simple, manageable and efficient regimen.

A phase I trial with recombinant interferon gamma (Roussel UCLAF) in advanced cancer patients.

A total of 29 patients with advanced malignancy were treated with recombinant interferon gamma (rIFN gamma, specific activity = 2.10(7) units/mg, purity greater than 95%) given by intravenous bolus at doses escalating from 0.01 mg/m2 to 5 mg/m2 (2 x 10(5) - 10(8) IU/m2) in nine successive steps (at least 3 patients/step). Injections of rIFN gamma were repeated every 72 h for 15 days. Toxicity was evaluated according to the WHO scale. Fever and chills occurred in all patients treated without clear dose effect. Nausea and vomiting appeared at the fifth dose level and their frequency seemed to be dose-related. Cardiovascular side-effects (first-degree atrioventricular reversible block) were observed at the 2 mg/m2 and 5 mg/m2 levels (3 patients). Hematological toxicities were mild (2 grade 1 and 1 grade II cases of granulocytopenia). Minor biological modifications included a transitory rise in hepatic enzymes (12 patients), which correlated with the presence of liver metastasis. Hypocholesterolemia was observed in 18 patients. The appearance of antibodies against rIFN gamma was not detected. One partial clinical response was observed in a patient receiving 2 mg/m2. During rIFN gamma therapy this patient had the highest scores in this series for peripheral T lymphocytes with an activated phenotype (HLA DR+, TAC+) = 15% and for natural killer (NK) cells (NKH1, Leu19+) = 17%. rIFN gamma appears as a well-tolerated and promising therapeutic agent with toxicities and mode of action probably distinct from IFN alpha and beta.

Phase II trial of methotrexate-FAM (m-FAM) in adenocarcinoma of unknown primary.

Nineteen patients with adenocarcinoma of unknown primary were treated with the m-FAM regimen, consisting of methotrexate 50 mg/m2 days 0, 28; 5-fluorouracil 600 mg/m2 days 1, 8, 29, 36; Adriamycin 30 mg/m2 days 1, 29; and mitomycin-C 10 mg/m2 day 1. All drugs were recycled every 56 days. No complete responses were seen. Seven patients (37%) achieved partial remission for a median duration of 11 months. An additional nine patients (47%) had stable disease for a median duration of 6 months. Median survival for responders was 16 months and was 10 months for those with stable disease. Toxicity was acceptable. This Phase II study attempted to evaluate the clinical impact of the pharmacological modulation of 5-fluorouracil with methotrexate, the goal being improvement of the results of FAM alone in adenocarcinoma of unknown primary. However, the addition of methotrexate, at least in the schedule employed in this study, did not appear superior to FAM.

Preoperative induction chemotherapy in the treatment of locally advanced soft tissue sarcomas.

Evidence that supports the use of systemic, presurgical induction chemotherapy to render soft tissue sarcomas resectable or to minimize the extent of surgical excision is presented. Induction chemotherapy was administered in 34 cases of nonmetastatic soft tissue sarcomas. All patients had large tumors for which only mutilating surgery, if any, was possible. In 21 patients, a combination of Adriamycin (doxorubicin), cyclophosphamide, cisplatin, vindesine, and DTIC (DCPAV) produced two complete remissions (CR) and 6 partial remissions (PR). A combination of cyclophosphamide, vincristine, Adriamycin, and DTIC (CYVADIC) produced three PR in eight patients, and a combination of Adriamycin and ifosfamide (AI) produced two PR in five patients. After two to seven cycles of chemotherapy, 24 patients underwent surgery. In 19, gross tumor excision was performed; 12 proved to be microscopically fully resected. Disease in two patients entered CR with chemotherapy alone, but surgery was performed in both patients as well. Irradiation was administered in ten patients to produce or insure CR (eight cases of residual disease postoperatively), and in two patients with unresectable disease. Four patients with disease in CR after surgery also received radiation due to the initial massive tumor size. The Kaplan-Meier survival curves at 2 years showed 18% total survival in the patients in whom CR of disease was not achieved, and 80% survival in patients with disease in CR. Of the 22 patients with disease in CR (by all means), disease-free survival was 1 to 44 months (mean, 13.7). Disease currently remains in CR in ten patients with a mean follow-up of 13.6 months (3 to 34 months from end of therapy). Ten patients had a local recurrence following a CR after 3 to 44 months (mean, 15.3 months).

[Soft tissue clear cell sarcomas. Reevaluation of clear cell sarcomas of the tendons and the aponeuroses in 14 cases]. / Les sarcomes à cellules claires des tissus mous. Réévaluation des sarcomes à cellules claires des tendons et des gaines à propos de 14 cas.

A retrospective study is realized including 14 cases of clear cell sarcomas of tendons and aponeuroses, found in the files of Institut Gustave-Roussy (1969-1983). The main microscopic features are emphasized. In comparison with other soft tissue sarcomas, clear cell sarcomas are clinically characterized by a great frequency of local lymph node metastases (4 of 14 cases). The prognosis is usually particularly poor. The exact histogenesis remains obscure, but the results of special stains, support origin from migrated neural crest cells: melanin is visualized in 2 out of 12 tumors in which Fontana-Masson is performed. S 100 protein, a neuroectodermal marker, is positive by immunoperoxidase technique (PAP) in 6 of 10 tested cases. A better diagnostic approach is allowed with the help of ultrastructural study performed in 3 cases. These tumors must be separated from synovialosarcomas. Because the histogenesis is not actually sure, it is preferable to call them "soft tissue clear cell sarcomas".

[Primary chemotherapy in soft tissue sarcomas considered inoperable]. / Chimiothérapie première dans les sarcomes des parties molles considérés comme inopérables.

Fourteen adult patients with inoperable soft tissue sarcoma (with metastases in 4 cases) received chemotherapy as primary treatment. Nine cases were treated by CYVADIC (cyclophosphamide, vincristin, doxorubicin, DTIC) and five cases by DECAV (DTIC, cyclophosphamide, cis-platinum, doxorubicin, vindesine). An objective response was obtained in 7 cases (1 complete remission and 6 regressions greater than 50%) and stabilization in 4 cases. Seven patients became operable after 2 to 6 courses of chemotherapy and a complete resection could be performed in 6 cases. The duration of the response was 2 to 39 months. Toxicity with both combinations was acceptable. We conclude that chemotherapy can provide initial tumor reduction and permit subsequently less radical surgery.