Multifunctional In-Memory Logics Based on a Dual-Gate Antiambipolar Transistor toward Non-von Neumann Computing Architecture.

In-memory computing may make it possible to realize non-von Neumann computing because the logic circuits are unified in the memory units. We investigated two types of in-memory logic operations, namely, two-input logic circuits and multifunctional artificial synapses. These were realized in a dual-gate antiambipolar transistor (AAT) with a ReS2/WSe2 heterojunction, in which polystyrene with a zinc phthalocyanine core (ZnPc-PS4) was incorporated as a memory layer. Here, reconfigurability is a key concept for both types of device operations and was achieved by merging the Λ-shaped transfer curve of the AAT and the nonvolatile memory effect of ZnPc-PS4. First, we achieved electrically reconfigurable two-input logic circuits. Versatile logic circuits such as AND, OR, NAND, NOR, and XOR circuits were demonstrated by taking advantage of the Λ-shaped transfer curve of the dual-gate AAT. Importantly, the nonvolatile memory function provided the electrical switching of the individual circuits between AND/OR, NAND/NOR, and XOR/NAND circuits with constant input signals. Second, the memory effect was applied to multifunctional artificial synapses. The inhibitory/excitatory and long-term potentiation/depression synaptic operations were electrically reconfigured simply by controlling one parameter (readout voltage), making three distinct responses possible even with the same presynaptic signals. These findings provide hints that may lead to the realization of new in-memory computing architectures beyond the current von Neumann computers.

Reconfigurable Logic-in-Memory Constructed Using an Organic Antiambipolar Transistor.

We demonstrate an electrically reconfigurable two-input logic-in-memory (LIM) using a dual-gate-type organic antiambipolar transistor (DG-OAAT). The attractive feature of this device is that a phthalocyanine-cored star-shaped polystyrene is used as a nano-floating gate, which enables the electrical switching of individual logic circuits and stores the circuit information by the nonvolatile memory effect. First, the DG-OAAT exhibited Λ-shaped transfer curves with hysteresis by sweeping the bottom-gate voltage. Programming and erasing operations enabled the reversible shift of the Λ-shaped transfer curves. Furthermore, the top-gate voltage effectively tuned the peak voltages of the transfer curves. Consequently, the combination of dual-gate and memory effects achieved electrically reconfigurable two-input LIM operations. Individual logic circuits (e.g., OR/NAND, XOR/NOR, and AND/XOR) were reconfigured by the corresponding programming and erasing operations without any variations in the input signals. Our device concept has the potential to fulfill an epoch-making organic integration circuit with a simple device configuration.

Optically Controllable Organic Logic-in-Memory: An Innovative Approach toward Ternary Data Processing and Storage.

Logic-in-memory (LIM) has emerged as an energy-efficient computing technology, as it integrates logic and memory operations in a single device architecture. Herein, a concept of ternary LIM is established. First, a p-type 2,7-dioctyl[1]benzothieno[3,2-b][1]benzothiophene (C8-BTBT) transistor is combined with an n-type PhC2H4-benzo[de]isoquinolino[1,8-gh]quinolone diimide (PhC2-BQQDI) transistor to obtain a binary memory inverter, in which a zinc phthalocyanine-cored polystyrene (ZnPc-PS4) layer serves as a floating gate. The contrasting photoresponse of the transistors toward visible and ultraviolet light and the efficient hole-trapping ability of ZnPc-PS4 enable us to achieve an optically controllable memory operation with a high memory window of 18 V. Then, a ternary memory inverter is developed using an anti-ambipolar transistor to achieve a three-level data processing and storage system for more advanced LIM applications. Finally, low-voltage operation of the devices is achieved by employing a high-k dielectric layer, which highlights the potential of the developed LIM units for next-generation low-power electronics.

Preparation of Supramolecular Miktoarm Star Copolymers with a Zinc Phthalocyanine Core through ATRP and RAFT Polymerization.

Topological polymers have attracted considerable attention owing to their unique chemical and physical properties. This study demonstrates the formation of novel supramolecular miktoarm star copolymers with a zinc phthalocyanine (ZnPc) core using metal-ligand coordination interactions. Various linear polymers with pyridyl end groups, poly(methyl methacrylate), poly(vinyl acetate) and poly(N-vinyl carbazole), are prepared via reversible addition-fragmentation chain transfer (RAFT) polymerization. This facilitates coordination to the ZnPc core of 4-armed star-shaped polystyrene prepared via atom-transfer radical polymerization (ATRP). Furthermore, the formation of a 1:1 complex of a ZnPc molecule and pyridyl group of the chain-transfer agent for RAFT is confirmed by absorption spectral studies and 1 H NMR spectroscopic analyses. The concept of supramolecular complexation can be extended to the preparation of AB4 -type supramolecular miktoarm star-shaped copolymers with functional cores.

Assessment of KRAS G12C Target Engagement by a Covalent Inhibitor in Tumor Biopsies Using an Ultra-Sensitive Immunoaffinity 2D-LC-MS/MS Approach.

KRAS is one of the most frequently mutated oncogenes, with KRAS G12C recently becoming an actionable target for small molecule intervention. GDC-6036 is an investigational KRAS G12C inhibitor that acts by irreversibly binding to the switch II pocket of KRAS G12C when in the inactive GDP-bound state, thereby blocking GTP binding and activation. Assessing target engagement is an essential component of clinical drug development, helping to demonstrate mechanistic activity, guide dose selection, understand pharmacodynamics as it relates to clinical response, and explore resistance. Here, we report the development of an ultra-sensitive approach for assessing KRAS G12C engagement. Immunoaffinity enrichment with a commercially available anti-RAS antibody was combined with a targeted 2D-LC-MS/MS technique to quantify both free and GDC-6036-bound KRAS G12C proteins. A KRAS G12C-positive non-small cell lung cancer xenograft model was dosed with GDC-6036 to assess the feasibility of this assay for analyzing small core needle biopsies. As predicted, dose-dependent KRAS G12C engagement was observed. To date, a sensitivity of 0.08 fmol/µg of total protein has been achieved for both free and GDC-6036-bound KRAS G12C with as little as 4 µg of total protein extracted from human tumor samples. This sub-fmol/µg level of sensitivity provides a powerful potential approach to assess covalent inhibitor target engagement at the site of action using core needle tumor biopsies from clinical studies.

Giredestrant reverses progesterone hypersensitivity driven by estrogen receptor mutations in breast cancer.

ESR1 (estrogen receptor 1) hotspot mutations are major contributors to therapeutic resistance in estrogen receptor-positive (ER+) breast cancer. Such mutations confer estrogen independence to ERα, providing a selective advantage in the presence of estrogen-depleting aromatase inhibitors. In addition, ESR1 mutations reduce the potency of tamoxifen and fulvestrant, therapies that bind ERα directly. These limitations, together with additional liabilities, inspired the development of the next generation of ERα-targeted therapeutics, of which giredestrant is a high-potential candidate. Here, we generated Esr1 mutant-expressing mammary gland models and leveraged patient-derived xenografts (PDXs) to investigate the biological properties of the ESR1 mutations and their sensitivity to giredestrant in vivo. In the mouse mammary gland, Esr1 mutations promote hypersensitivity to progesterone, triggering pregnancy-like tissue remodeling and profoundly elevated proliferation. These effects were driven by an altered progesterone transcriptional response and underpinned by gained sites of ERα-PR (progesterone receptor) cobinding at the promoter regions of pro-proliferation genes. PDX experiments showed that the mutant ERα-PR proliferative program is also relevant in human cancer cells. Giredestrant suppressed the mutant ERα-PR proliferation in the mammary gland more so than the standard-of-care agents, tamoxifen and fulvestrant. Giredestrant was also efficacious against the progesterone-stimulated growth of ESR1 mutant PDX models. In addition, giredestrant demonstrated activity against a molecularly characterized ESR1 mutant tumor from a patient enrolled in a phase 1 clinical trial. Together, these data suggest that mutant ERα can collaborate with PR to drive protumorigenic proliferation but remain sensitive to inhibition by giredestrant.

Electroactive Soft Actuators Based on Columnar Ionic Liquid Crystal/Polymer Composite Membrane Electrolytes Forming 3D Continuous Ionic Channels.

Here, we report low-voltage-driven fast-response nanostructured columnar ionic liquid crystal/polymer composite actuators that form three-dimensional continuous ion channels. A three-component self-assembly of a zwitterionic rod-like molecule (49.5 wt %), an ionic liquid (27.5 wt %), and poly(vinyl alcohol) (23.0 wt %) provided a free-standing stretchable membrane electrolyte. The dissociated ions can move through a continuous 3D ionophilic matrix surrounding the hydrophobic columns formed by the hexagonally organized rod-mesogens. Three-layer actuators composed of the electrolyte film sandwiched between two conductive polymer film electrodes of doped polythiophene exhibited a bending motion with 0.32% strain and moved 2 mm within 220 ms under 1 V at 0.1 Hz in 70% relative humidity due to the formation of electric double layers at the soft solid electrolyte/electrode interfaces. The bending strain of the columnar nanostructured actuator is comparable to those of polymer iongel actuators and block polymer actuators containing 25-80 wt % of ionic liquids. It is noteworthy that a small number of ions organized into the 3D nanochannels can generate the large bending deformation, which can contribute to reduce the risk of leakage of ions and the production cost. In addition, we have demonstrated a low-voltage-driven deformable mirror actuator that is expected to be applied to optical devices.

Miktoarm Star Copolymers Prepared by Transformation from Enhanced Spin Capturing Polymerization to Nitroxide-Mediated Polymerization (ESCP-T-NMP) toward Nanomaterials.

Reversible-deactivation radical polymerization (RDRP) serves as a powerful tool nowadays for the preparations of unique linear and non-linear macromolecules. In this study, enhanced spin capturing polymerizations (ESCPs) of styrene (St) and tert-butyl acrylate (tBA) monomers were, respectively, conducted in the presence of difunctional (1Z,1'Z)-1,1'-(1,4-phenylene) bis (N-tert-butylmethanimine oxide) (PBBN) nitrone. Four-arm (PSt)4 and (PtBA)4 star macroinitiators (MIs) can be afforded. By correspondingly switching the second monomer (i.e., tBA and St), miktoarm star copolymers (µ-stars) of (PSt)2-µ-(PtBA-b-PSt)2 and (PtBA)2-µ-(PSt-b-PtBA)2) were thus obtained. We further conducted hydrolysis of the PtBA segments to PAA (i.e., poly(acrylic acid)) in µ-stars to afford amphiphilic µ-stars of (PSt)2-µ-(PAA-b-PSt)2 and (PAA)2-µ-(PSt-b-PAA)2. We investigated each polymerization step and characterized the obtained two sets of "sequence-isomeric" µ-stars by FT-IR, 1H NMR, differential scanning calorimeter (DSC), and thermogravimetric analysis (TGA). Interestingly, we identified their physical property differences in the case of amphiphilic µ-stars by water contact angle (WCA) and atomic force microscopy (AFM) measurements. We thus proposed two microstructures caused by the difference of polymer chain sequences. Through this polymerization transformation (T) approach (i.e., ESCP-T-NMP), we demonstrated an interesting and facile strategy for the preparations of µ-stars with adjustable/switchable interior and exterior polymer structures toward the preparations of various nanomaterials.

Low-pass Whole-genome Sequencing of Circulating Cell-free DNA Demonstrates Dynamic Changes in Genomic Copy Number in a Squamous Lung Cancer Clinical Cohort.

PURPOSE: We developed a method to monitor copy number variations (CNV) in plasma cell-free DNA (cfDNA) from patients with metastatic squamous non-small cell lung cancer (NSCLC). We aimed to explore the association between tumor-derived cfDNA and clinical outcomes, and sought CNVs that may suggest potential resistance mechanisms. EXPERIMENTAL DESIGN: Sensitivity and specificity of low-pass whole-genome sequencing (LP-WGS) were first determined using cell line DNA and cfDNA. LP-WGS was performed on baseline and longitudinal cfDNA of 152 patients with squamous NSCLC treated with chemotherapy, or in combination with pictilisib, a pan-PI3K inhibitor. cfDNA tumor fraction and detected CNVs were analyzed in association with clinical outcomes. RESULTS: LP-WGS successfully detected CNVs in cfDNA with tumor fraction ≥10%, which represented approximately 30% of the first-line NSCLC patients in this study. The most frequent CNVs were gains in chromosome 3q, which harbors the PIK3CA and SOX2 oncogenes. The CNV landscape in cfDNA with a high tumor fraction generally matched that of corresponding tumor tissue. Tumor fraction in cfDNA was dynamic during treatment, and increases in tumor fraction and corresponding CNVs could be detected before radiographic progression in 7 of 12 patients. Recurrent CNVs, such as MYC amplification, were enriched in cfDNA from posttreatment samples compared with the baseline, suggesting a potential resistance mechanism to pictilisib. CONCLUSIONS: LP-WGS offers an unbiased and high-throughput way to investigate CNVs and tumor fraction in cfDNA of patients with cancer. It may also be valuable for monitoring treatment response, detecting disease progression early, and identifying emergent clones associated with therapeutic resistance.

Genomic Alterations Associated with Recurrence and TNBC Subtype in High-Risk Early Breast Cancers.

The identification of early breast cancer patients who may benefit from adjuvant chemotherapy has evolved to include assessment of clinicopathologic features such as tumor size and nodal status, as well as several gene-expression profiles for ER-positive, HER2-negative cancers. However, these tools do not reliably identify patients at the greatest risk of recurrence. The mutation and copy-number landscape of triple-negative breast cancer (TNBC) subtypes defined by gene expression is also largely unknown, and elucidation of this landscape may shed light on novel therapeutic opportunities. The USO01062 phase III clinical trial of standard chemotherapy (with or without capecitabine) enrolled a cohort of putatively high-risk patients based on clinical features, yet only observed a 5-year disease-free survival event rate of 11.6%. In order to uncover genomic aberrations associated with recurrence, a targeted next-generation sequencing panel was used to compare tumor specimens from patients who had a recurrence event with a matched set who did not. The somatic mutation and copy-number alteration landscapes of high-risk early breast cancer patients were characterized and alterations associated with relapse were identified. Tumor mutational burden was evaluated but was not prognostic in this study, nor did it correlate with PDL1 or CD8 gene expression. However, TNBC subtypes had substantial genomic heterogeneity with a distinct pattern of genomic alterations and putative underlying driver mutations. IMPLICATIONS: The present study uncovers a compendium of genomic alterations with utility to more precisely identify high-risk patients for adjuvant trials of novel therapeutic agents.

Synthesis of Poly(ε-caprolactone)-Based Miktoarm Star Copolymers through ROP, SA ATRC, and ATRP.

The synthesis of novel branched/star copolymers which possess unique physical properties is highly desirable. Herein, a novel strategy was demonstrated to synthesize poly(ε-caprolactone) (PCL) based miktoarm star (µ-star) copolymers by combining ring-opening polymerization (ROP), styrenics-assisted atom transfer radical coupling (SA ATRC), and atom transfer radical polymerization (ATRP). From the analyses of gel permeation chromatography (GPC), proton nuclear magnetic resonance (¹H NMR), and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), well-defined PCL-µ-PSt (PSt: polystyrene), and PCL-µ-PtBA (PtBA: poly(tert-butyl acrylate) µ-star copolymers were successfully obtained. By using atomic force microscopy (AFM), interestingly, our preliminary examinations of the µ-star copolymers showed a spherical structure with diameters of ca. 250 and 45 nm, respectively. We successfully employed combinations of synthetic techniques including ROP, SA ATRC, and ATRP with high effectiveness to synthesize PCL-based µ-star copolymers.

Synthesis of Novel µ-Star Copolymers with Poly(N-Octyl Benzamide) and Poly(ε-Caprolactone) Miktoarms through Chain-Growth Condensation Polymerization, Styrenics-Assisted Atom Transfer Radical Coupling, and Ring-Opening Polymerization.

Star copolymers are known to phase separate on the nanoscale, providing useful self-assembled morphologies. In this study, the authors investigate synthesis and assembly behavior of miktoarm star (µ-star) copolymers. The authors employ a new strategy for the synthesis of unprecedented µ-star copolymers presenting poly(N-octyl benzamide) (PBA) and poly(ε-caprolactone) (PCL) arms: a combination of chain-growth condensation polymerization, styrenics-assisted atom transfer radical coupling, and ring-opening polymerization. Gel permeation chromatography, mass-analyzed laser desorption/ionization mass spectrometry, and 1 H NMR spectroscopy reveal the successful synthesis of a well-defined (PBA11 )2 -(PCL15 )4 µ-star copolymer (Mn,NMR ≈ 12 620; D = 1.22). Preliminary examination of the PBA2 PCL4 µ-star copolymer reveals assembled nanofibers having a uniform diameter of ≈20 nm.

Heterogeneity and clinical significance of ESR1 mutations in ER-positive metastatic breast cancer patients receiving fulvestrant.

Mutations in ESR1 have been associated with resistance to aromatase inhibitor (AI) therapy in patients with ER+ metastatic breast cancer. Little is known of the impact of these mutations in patients receiving selective oestrogen receptor degrader (SERD) therapy. In this study, hotspot mutations in ESR1 and PIK3CA from ctDNA were assayed in clinical trial samples from ER+ metastatic breast cancer patients randomized either to the SERD fulvestrant or fulvestrant plus a pan-PI3K inhibitor. ESR1 mutations are present in 37% of baseline samples and are enriched in patients with luminal A and PIK3CA-mutated tumours. ESR1 mutations are often polyclonal and longitudinal analysis shows distinct clones exhibiting divergent behaviour over time. ESR1 mutation allele frequency does not show a consistent pattern of increases during fulvestrant treatment, and progression-free survival is not different in patients with ESR1 mutations compared with wild-type patients. ESR1 mutations are not associated with clinical resistance to fulvestrant in this study.

Picket-fence polythiophene and its diblock copolymers that afford microphase separations comprising a stacked and an isolated polythiophene ensemble.

All-polythiophene diblock copolymers, comprising one unsheathed block and one fenced block, were synthesized through catalyst-transfer polycondensation. The unsheathed block self-assembles through π-π stacking, thereby inducing microphase separation. Consequently, we have succeeded in creating a microphase separation comprising an ensemble of stacked and isolated polythiophenes. This achievement could be extended to various unexplored applications as a result of the integration of the contrasting functions of the two blocks.

Nonvolatile liquid anthracenes for facile full-colour luminescence tuning at single blue-light excitation.

Nonvolatile room-temperature luminescent molecular liquids are a new generation of organic soft materials. They possess high stability, versatile optical properties, solvent-free fluid behaviour and can effectively accommodate dopant dye molecules. Here we introduce an approach to optimize anthracene-based liquid materials, focussing on enhanced stability, fluorescence quantum yield, colour tunability and processability, with a view to flexible electronic applications. Enveloping the anthracene core in low-viscosity branched aliphatic chains results in stable, nonvolatile, emissive liquid materials. Up to 96% efficient energy-transfer-assisted tunable emission is achieved by doping a minute amount of acceptor dye in the solvent-free state. Furthermore, we use a thermoresponsive dopant to impart thermally controllable luminescence colours. The introduced strategy leading to diverse luminescence colours at a single blue-light excitation can be an innovative replacement for currently used luminescent materials, providing useful continuous emissive layers in developing foldable devices.

A first in human phase 1 study of CG0070, a GM-CSF expressing oncolytic adenovirus, for the treatment of nonmuscle invasive bladder cancer.

PURPOSE: We assessed the safety, pharmacokinetics and anticancer activity of intravesical CG0070, a cancer selective, replication competent adenovirus, for the treatment of nonmuscle invasive bladder cancer. MATERIALS AND METHODS: A total of 35 patients received single or multiple (every 28 days × 3 or weekly × 6) intravesical infusions of CG0070 at 1 of 4 dose levels (1 × 10(12), 3 × 10(12), 1 × 10(13) or 3 × 10(13) viral particles). Response to treatment was based on cystoscopic assessment and biopsy or urine cytology. Urine and plasma CG0070, and granulocyte-monocyte colony-stimulating factor were measured in all patients. A subset of 18 patients was assessed for retinoblastoma phosphorylation status. RESULTS: Grade 1-2 bladder toxicities were the most common adverse events observed. A maximum tolerated dose was not reached. High levels of granulocyte-monocyte colony-stimulating factor were detected in urine after administration in all patients. Virus replication was suggested based on an increase in urine CG0070 genomes between days 2 and 5 in 58.3% of tested patients (7 of 12). The complete response rate and median duration of the complete response across cohorts was 48.6% and 10.4 months, respectively. In the multidose cohorts the complete response rate for the combined groups (every 28 days and weekly × 6) was 63.6% (14 of 22 patients). In an exploratory, retrospective assessment patients with borderline or high retinoblastoma phosphorylation who received the multidose schedules had an 81.8% complete response rate (9 of 11). CONCLUSIONS: Intravesical CG0070 was associated with a tolerable safety profile and antibladder cancer activity. Granulocyte-monocyte colony-stimulating factor transgene expression and CG0070 replication were also suggested.

Solvent-free luminescent organic liquids.

Illuminating! Isolation of a π-core by covalently attached flexible hydrocarbon chains has been employed to synthesize blue-emitting oligo(p-phenylenevinylene) (OPV) liquids with tunable viscosity and optical properties. A solvent-free, stable, white-light emitting ink/paint, which can be applied onto various surfaces and even onto LEDs, was made by blending of liquid OPVs with emissive solid dopants.

Excitation energy migration processes in self-assembled porphyrin boxes constructed by conjugated porphyrin dimers.

meso-Pyridine-appended alkynylene-bridged zinc(II) porphyrin dimers D2 and D4 assemble spontaneously, in noncoordinating solvents such as toluene, into tetrameric porphyrin boxes B2 and B4, respectively. Interestingly, the formation of Bn from Dn leads to the two kinds of self-assembled porphyrin boxes constructed by planar and orthogonal conformers, respectively. Excitation energy migration processes within these assemblies have been investigated in detail by using both steady-state and time-resolved spectroscopic methods. The pump-power dependence on the femtosecond transient absorption decay profiles is directly associated with the excitation energy migration process within the Bn boxes, where the exciton-exciton annihilation time is well-described in terms of the Foster-type incoherent energy hopping model. Consequently, the excitation energy hopping rates in porphyrin boxes constructed by planar and orthogonal conformers have been estimated to be (approximately 1.2 ps)(-1) and (approximately 1 ps)(-1), respectively. Furthermore, the porphyrin boxes constructed by orthogonal conformers show additional slow excitation energy hopping rates of (approximately 12 ps)(-1). Overall, the dihedral angle in the constituent dimers is a key factor to control the energy transfer efficiency for the fabrication of artificial light-harvesting complexes using conjugated porphyrin dimer systems.

Use of a bioamplification assay to detect nonselective recombinants and assess the genetic stability of oncolytic adenoviruses.

Detection of nonselective adenoviruses in tissue- or tumor-selective oncolytic adenovirus preparations presents a technical challenge because of the conditionally replication-competent nature of oncolytic adenoviruses. Although quantitative PCR has been used extensively for detecting specific genes that are likely present in nonselective recombinants, the actual biological activity of nonselective genetic recombinants has not been demonstrated. Therefore, a bioassay that amplifies nonselective adenoviruses through multiple passages in nonpermissive cells was developed to detect biologically active nonselective recombinants using CG7870, a prostate-specific oncolytic adenovirus. The assay was sensitive, and its results were consistent with a quantitative PCR assay for four lots of CG7870. CG0070, a pan-tumor oncolytic adenovirus with no detectable wild-type-like recombinants by PCR, was subjected to a variation of this bioamplification assay using two different nonpermissive cell lines to both verify PCR results and assess its genetic stability under selection pressure. No evidence of the presence of biologically active nonselective recombinants was seen in the original material or after serial passaging in nonpermissive cells. Thus, this bioamplification assay is able to detect nonselective recombinants, and its results are consistent with quantitative PCR assays. A modified version of this assay is also useful for assessing the genetic stability of oncolytic adenoviruses that have no PCR-detectable recombinants.

Phase 1/2 dose-escalation study of a GM-CSF-secreting, allogeneic, cellular immunotherapy for metastatic hormone-refractory prostate cancer.

BACKGROUND: This open-label, multicenter, dose-escalation study evaluated multiple dose levels of immunotherapy in patients with metastatic hormone-refractory prostate cancer (HRPC). The immunotherapy, based on the GVAX platform, consisted of 2 allogeneic prostate-carcinoma cell lines modified to secrete granulocyte-macrophage-colony-stimulating factor (GM-CSF). METHODS: Dose levels ranged from 100 x 10(6) cells q28d x 6 to 500 x 10(6) cells prime/300 x 10(6) cells boost q14d x 11. Endpoints included safety, immunogenicity, overall survival, radiologic response, prostate-specific antigen (PSA) kinetics, and serum GM-CSF pharmacokinetics. RESULTS: Eighty men, median age 69 years (range, 49-90 years), were treated. The most common adverse effect was injection-site erythema. Overall, the immunotherapy was well tolerated. A maximal tolerated dose was not established. The median survival time was 35.0 months in the high-dose group, 20.0 months in the mid-dose, group, and 23.1 months in the low-dose group. PSA stabilization occurred in 15 (19%) patients, and a >50% decline in PSA was seen in 1 patient. The proportion of patients who generated an antibody response to 1 or both cell lines increased with dose and included 10 of 23 (43%) in the low-dose group, 13 of 18 (72%) in the mid-dose group, and 16 of 18 (89%) in the high-dose group (P = .002; Cochran-Armitage trend test). CONCLUSIONS: This immunotherapy was well tolerated. Immunogenicity and overall survival varied by dose. Two phase 3 trials in patients with metastatic HRPC are underway.