Heterogeneous Response Research Articles

Cisplatin-Induced Metabolic Responses Measured with Raman Spectroscopy in Cancer Cells, Spheroids, and Canine-Derived Organoids.

Ex vivo assessment of drug response with conventional cell viability assays remains the standard practice for guiding initial therapeutic choices. However, such ensemble approaches fail to capture heterogeneities in treatment response and cannot identify early markers of response. Here, we leverage Raman spectroscopy (RS) as an accurate, low-cost, extraction-free, and label-free approach to track metabolic changes in cancer cells, spheroids, and organoids in response to cisplatin treatment. We identified 12 statistically significant metabolites in cells and 19 metabolites in spheroids and organoids as a function of depth. We show that the cisplatin treatment of 4T1 cells and spheroids results in a shift in metabolite levels; metabolites including nucleic acids such as DNA, 783 cm-1 with p = 0.00021 for cells; p = 0.02173 for spheroids, major amino acids such as threonine, 1338 cm-1 with p = 0.00045 for cells; p = 0.01022 for spheroids, proteins such as amide III, 1248 cm-1 with p = 0.00606 for cells; p = 0.00511 for spheroids serve as early predictors of response. Our RS findings were also applicable to canine-derived organoids, showing spatial variations in metabolic changes as a function of organoid depth in response to cisplatin. Further, the metabolic pathways such as tricarboxylic acid (TCA)/citric acid cycle and glyoxylate and dicarboxylate metabolism that drive drug response showed significant differences based on organoid depth, replicating the heterogeneous treatment response seen in solid tumors where there is a difference from the periphery to the tumor core. Our study showcases the versatility of RS as a predictive tool for treatment response applicable from cells to organotypic cultures, that has the potential to decrease animal burden and readout time for preclinical drug efficacy.

Combining price and non-price interventions for water conservation

Abstract Marginal pricing has long been the instrument of choice to address water conservation challenges. More recently, non-price behavioral interventions have emerged as an alternative. However, there is limited data on the relative efficacies of price and non-price interventions. We report results from long-term field experiments studying unit-level water conservation responses to both price and non-price interventions in the same group of households (n = 64 186 household-days). Conservation habits, attitude-action gaps, principal-agent incongruities, and billing cycles help account for the heterogeneity in response between households, and across time. A non-price behavior modification intervention before the introduction of marginal pricing resulted in a large and significant effect on treated households (33%). The subsequent introduction of marginal volumetric pricing also reduced water use (8%, for previously untreated households). However, this average price effect masks how a large share (21%) of households increased water use, or how a mere 12% of the households accounted for all the aggregate reduction in water use. We investigated such heterogeneous responses as a systematic conservation maximization design question beyond statistical variance in individual responses. We used daily water consumption measurements across three years alongside a household survey to delineate structural and agentic barriers to conservation behavior. Our analysis reveals how combining price and non-price behavioral interventions could hold the key to achieving conservation effects that are both large and persistent.

Long-term mesoscale imaging of 3D intercellular dynamics across a mammalian organ

A comprehensive understanding of physio-pathological processes necessitates non-invasive intravital three-dimensional (3D) imaging over varying spatial and temporal scales. However, huge data throughput, optical heterogeneity, surface irregularity, and phototoxicity pose great challenges, leading to an inevitable trade-off between volume size, resolution, speed, sample health, and system complexity. Here, we introduce a compact real-time, ultra-large-scale, high-resolution 3D mesoscope (RUSH3D), achieving uniform resolutions of 2.6×2.6×6μm3 across a volume of 8,000×6,000×400μm3 at 20Hz with low phototoxicity. Through the integration of multiple computational imaging techniques, RUSH3D facilitates a 13-fold improvement in data throughput and an orders-of-magnitude reduction in system size and cost. With these advantages, we observed premovement neural activity and cross-day visual representational drift across the mouse cortex, the formation and progression of multiple germinal centers in mouse inguinal lymph nodes, and heterogeneous immune responses following traumatic brain injury-all at single-cell resolution, opening up a horizon for intravital mesoscale study of large-scale intercellular interactions at the organ level.

Partisan differences in perceived levels of democracy across presidential administrations

Democracy in the United States is in decline. The current work examines to what degree U.S. Americans perceive this decline in the U.S., and how partisanship may shape those perceptions over time. Using cross-sectional archival data spanning three U.S. presidential administrations (Study 1), we find that perceived levels of democracy by U.S. Americans on both sides of the political spectrum were not attuned to objective democracy indices but instead followed a partisan gap where election winners perceived higher levels of democracy than election losers. Using a longitudinal quasi-experiment tracking the 2020 presidential election (Study 2), we find that the effect of partisanship on perceived levels of democracy kicked in right after the announcement of the election winner but was the strongest after the official inauguration. Moreover, Trump supporters showed heterogeneous responses to the Capitol attack, which highlights the possibility of bipartisan willingness to defend democracy after salient attacks on democratic principles. We discuss the implications of the effect of partisanship on perceptions of democracy.

Exploring metabolic anomalies in COVID-19 and post-COVID-19: a machine learning approach with explainable artificial intelligence.

The COVID-19 pandemic, caused by SARS-CoV-2, has led to significant challenges worldwide, including diverse clinical outcomes and prolonged post-recovery symptoms known as Long COVID or Post-COVID-19 syndrome. Emerging evidence suggests a crucial role of metabolic reprogramming in the infection's long-term consequences. This study employs a novel approach utilizing machine learning (ML) and explainable artificial intelligence (XAI) to analyze metabolic alterations in COVID-19 and Post-COVID-19 patients. Samples were taken from a cohort of 142 COVID-19, 48 Post-COVID-19, and 38 control patients, comprising 111 identified metabolites. Traditional analysis methods, like PCA and PLS-DA, were compared with ML techniques, particularly eXtreme Gradient Boosting (XGBoost) enhanced by SHAP (SHapley Additive exPlanations) values for explainability. XGBoost, combined with SHAP, outperformed traditional methods, demonstrating superior predictive performance and providing new insights into the metabolic basis of the disease's progression and aftermath. The analysis revealed metabolomic subgroups within the COVID-19 and Post-COVID-19 conditions, suggesting heterogeneous metabolic responses to the infection and its long-term impacts. Key metabolic signatures in Post-COVID-19 include taurine, glutamine, alpha-Ketoglutaric acid, and LysoPC a C16:0. This study highlights the potential of integrating ML and XAI for a fine-grained description in metabolomics research, offering a more detailed understanding of metabolic anomalies in COVID-19 and Post-COVID-19 conditions.

Unveiling osmoregulation and immunological adaptations in Eleutheronema tetradactylum gills through high-throughput single-cell transcriptome sequencing

The fourfinger threadfin fish (Eleutheronema tetradactylum) is an economically significant species renowned for its ability to adapt to varying salinity environments, with gills serving as their primary organs for osmoregulation and immune defense. Previous studies focused on tissue and morphological levels, whereas ignored the cellular heterogeneity and the crucial gene information related to core cell subsets within E. tetradactylum gills. In this study, we utilized high-throughput single-cell RNA sequencing (scRNA-seq) to analyze the gills of E. tetradactylum, characterizing 16 distinct cell types and identifying unique gene markers and enriched functions associated within each cell type. Additionally, we subdivided ionocyte cells into four distinct subpopulations for the first time in E. tetradactylum gills. By employing weighted gene co-expression network analysis (WGCNA), we further investigated the cellular heterogeneity and specific response mechanisms to salinity fluctuant. Our findings revealed the intricate osmoregulation and immune functions of gill cells, highlighting their crucial roles in maintaining homeostasis and adapting to fluctuating salinity levels. This comprehensive cell-type atlas provides valuable insights into the species adaptive strategies, contributing to the conservation and management of this commercially significant fish as well as other euryhaline species.

A Third COVID-19 Vaccine Dose in Kidney Transplant Recipients Induces Antibody Response to Vaccine and Omicron Variants but Shows Limited Ig Subclass Switching.

This study addresses the challenges faced by kidney transplant recipients (KTRs) in mounting effective immune responses against COVID-19. By evaluating the antibody responses to a third dose of monovalent mRNA COVID-19 vaccine and its effectiveness against Omicron subvariants (BA.1 and BA.5), this study reveals significant reductions in both binding and neutralizing antibodies in KTRs compared to healthy controls. The research highlights altered IgG subclass switching and heterogeneous responses within the KTR population. Reduced recognition of variants, coupled with differences in IgG subclasses, decreases both the quality and quantity of protective antibodies after vaccination in KTRs. These findings underscore the need for tailored vaccination strategies for immunosuppressed populations such as KTRs. Alternative formulations and doses of COVID-19 vaccines should be considered for people with severely compromised immune systems, as more frequent vaccinations may not significantly improve the response, especially regarding neutralizing antibodies.

Humoral immune response as an indicator for protection against Covid-19 after anti-SARS-COV2-booster vaccination in hematological and oncological patients.

Cancer patients are at a higher risk to develop severe COVID-19 symptoms after SARS-CoV-2 infection compared to the general population and regularly show an impaired immune response to SARS-CoV-2 vaccination. In our oncological center, 357 patients with hematological and oncological diseases were monitored for neutralizing antibodies from October 2021 over 12 months. All patients had received three anti-SARS-CoV-2 vaccinations with an mRNA-(Comirnaty/BionTech or Spikevax/Moderna) or a vector vaccine (Vakzevria/AstraZeneca or JCOVDEN/Johnson&Johnson). Neutralizing anti-SARS-CoV-2 IgG antibodies in the patients' sera were detected within 3 months before, 3-10 weeks and 5-7 months after the booster vaccination (third vaccination). 112 patients developed a breakthrough SARS-CoV-2 infection during the observation period. High anti-SARS-Cov-2 antibody levels before infection significantly protected against symptomatic Covid-19 disease (p = .003). The median antibody titer in patients with asymptomatic Covid-19 disease was 2080 BAU/ml (binding antibody units per Milliliter) and 765 BAU/ml in symptomatic patients. 98% of the solid tumor patients reached seroconversion after the booster vaccination in comparison to 79% of the hematological patients. High antibody titers of >2080 BAU/ml after the booster vaccination were detected in 61% of the oncological and 34.8% of the hematological patients. 7-10 months after the booster vaccination, the anti-SARS-CoV-2 antibody titer declined to an average of 849 BAU/ml. Considering the heterogenous humoral immune response of cancer patients observed in this study, an individual vaccination strategy based on regular measurement of anti-SARS-CoV-2 antibody levels should be considered in contrast to fixed vaccination intervals.

Do Physicians Improve More from Positive or Negative Feedback?

We use clinical data on more than 240,000 surgeries and quasi-experimental methods to examine how physicians respond to the surprise release of a performance “report card.” Such feedback interventions are commonly used to encourage physicians to improve performance yet show limited evidence of success. Our results show that these limited effects mask heterogeneous behavioral responses to feedback valence. In particular, physicians improve more from positive feedback than from negative feedback, with negative feedback even reducing performance for a nontrivial share of patients. Experiments with laypersons replicate these results and show that struggles with negative feedback can be mitigated by giving incentives directly tied to improvement and by adding qualitative information that helps individuals interpret past performance. These results are consistent with behavioral models that suggest cognitive and emotional difficulties limit how well individuals use negative feedback. Thus, feedback interventions in healthcare should be carefully designed to mitigate these counterproductive behavioral responses. This paper was accepted by Ranjani Krishnan, accounting. Funding: We acknowledge generous funding provided by Tilburg University and the University of Massachusetts Amherst to run the laboratory experiments. Supplemental Material: The online appendices and data files are available at https://doi.org/10.1287/mnsc.2023.01340 .

Effects of property value assessment limit portability on migration outflows

AbstractThis study examines the impact of the 2008 portability provision of Florida's property value assessment limit, which permits homeowners to transfer the benefit of assessment limitations, on in‐state out‐migration. Leveraging IRS county‐to‐county migration data and quasi‐experimental designs, we find increased household and individual in‐state, county‐to‐county outflows following portability implementation in Florida relative to other states by 478 households (4.8%) and 675 individuals (3.5%). Our results suggest that natural amenities, rather than home‐ownership rates or property value growth, drive migration outflows. Robustness checks reveal heterogeneous responses across metro and nonmetro counties, suggesting that urbanization may accelerate after adopting the provision.

Heterogeneous response of estimated insulin sensitivity indices to metformin in young individuals with type 1 diabetes and different phenotypes

AimsThis study aimed to investigate whether the response to adding metformin to insulin in young adults with type 1 diabetes (T1D) differs according to weight phenotype and insulin sensitivity index.MethodsA prospective pilot study was conducted over 26 weeks in which insulin plus metformin (2 g/day) was administered to 35 individuals, ranging from normal weight (NW) to overweight (OW) to obese (OB) T1D individuals, to correlate insulin sensitivity indices and other clinical variables.ResultsAt the end of the follow-up period, all groups showed an increase in the eGDR (NW: 7.37 vs 8.16, p = 0.002; OW: 7.28 vs 8.24, p < 0.001; OB: 6.33 vs 7.52 p < 0.001). KITT and SEARCH SCORE improved only in the OB group (2.15 vs 3.14, p < 0.001 and 5.26 vs 5.72, p = 0.007, respectively). Furthermore, HbA1c and BMI were significantly greater in the OB group (− 0.62%, p < 0.001; − 1.12 kg/m2, p = 0.031, respectively). Regression analysis revealed that the serum levels of triglycerides and uric acid were significantly (0.059, p = 0.013; 0.076, p = 0.001) associated with insulin sensitivity indices.ConclusionsThe study showed that eGDR improved independently of basal weight after metformin treatment. However, the KITT and SEARCH indices improved only in the obese group. Triglycerides and uric acid are associated with insulin sensitivity indices. These results highlight the heterogeneity of the mechanisms underlying insulin resistance and its response to metformin in individuals with T1D.

T cells promote distinct transcriptional programs of cutaneous inflammatory disease in human skin structural cells.

T cells and structural cells coordinate appropriate inflammatory responses and restoration of barrier integrity following insult. Dysfunctional T cell activity precipitates tissue pathology that occurs alongside disease-associated alterations of structural cell subsets, but the mechanisms by which T cells promote these changes remain unclear. We show that subsets of circulating and skin-resident CD4+ T cells promote distinct transcriptional outcomes in human keratinocytes and dermal fibroblasts that correspond with divergent T cell cytokine production. Using these transcriptional signatures, we identify T cell-dependent outcomes associated with inflammatory skin disease, including a set of Th17 cell-induced genes in keratinocytes that are enriched in the skin during psoriasis and normalized by anti-IL-17 therapy, and a skin-resident T cell-induced gene module enriched in scleroderma-associated fibroblasts. Interrogating clinical data using T cell-derived structural cell gene networks enables investigation of the immune-dependent contribution to inflammatory disease and the heterogeneous patient response to biologic therapy.

Effects of plant health protecting product applications on the root-associated microbiota of apple saplings and strawberries

The root-associated microbiome has been of keen research interest due to its large potential for increasing plant performance. In agricultural systems, the use of plant health protecting products (PHPP) is essential for obtaining high yields and quality. Knowledge about the impact of PHPP applications on the root-associated microbiome remains limited, especially when applied above ground. We hypothesized that PHPP application leads to specific plant-mediated responses in the rhizosphere microbiota. We applied different PHPPs on greenhouse-grown apple and strawberry plants, cultivated in different soils. The effects of above ground PHPP applications on the root-associated microbiota were analyzed by 16S rRNA gene amplicon sequencing. No consistent effects were observed across three experimental trials, though alpha diversity tended to be decreased after PHPP application. In some cases, shifts in community composition became evident (PERMANOVA R2 = 0.192, p = 0.001) or increases in dispersion (PERMDISP Padj = 0.040). Movento® treatments caused in part deterministic shifts in community assembly, whereas Serenade®ASO treatments caused rather stochastic shifts. Effects were concluded to be probably transient and to disappear without continuous product application. These rather heterogenous temporary responses are in line with the concept of the ‘Anna Karenina Principle’, under which terms PHPP applications can act as mild stress factors and slightly and transiently alter the below ground bacterial community. These findings indicate that above ground PHPP applications can have product specific implications for the root-associated microbiome, which should be considered in the context of microbiome management strategies.

Immune-inflammatory markers and clinical characteristics as predictors of the depth of response and prognosis of patients with PD-L1 ≥50% metastatic non-small cell lung cancer receiving first-line immunotherapy.

Patients with programmed cell death-ligand 1 (PD-L1) ≥50% metastatic non-small cell lung cancer (NSCLC) treated with first-line immunotherapy showed heterogeneous tumor responses. In this study, we investigated the clinical and immune-inflammatory markers distinguishing patients with metastatic NSCLC achieving high depth of tumor response (HDPR) from those with non-high depth of response (NHDPR). The impact of clinical features on the prognosis of patients with PD-L1 ≥50% were further clarified. The clinical characteristics and immune-inflammatory markers of 17 patients with PD-L1 ≥50% metastatic NSCLC at Beijing Tiantan Hospital between July 2020 and December 2023 were retrospectively analyzed. Among the 17 patients, seven (41.2%) patients achieved HDPR (range: -50%, -72%) and 10 (58.8%) patients achieved NHDPR (range: -13%, -45%). Below normal CD4 + T lymphocytes/CD8 + T lymphocytes (CD4/CD8) ratio (p = 0.01) and oncogenes and/or tumor suppressor gene mutations (TP53/KRAS/EGFR) (p = 0.001) were found enriched for NHDPR compared with HDPR. With a median follow-up of 26.0 months (range: 17.2-34.8 months), the median progression-free survival (PFS) following first-line immunotherapy and overall survival (OS) were 9.0 months (95% CI: 5.0-13.0) and not reached (NR), respectively. The neutrophil-to-lymphocyte ratio (NLR) was identified as an independent prognostic factor on first-line PFS. Patients with an NLR ≥4 exhibited a shorter median PFS (7.0 months vs. NR; p = 0.033; 95% CI: 1.2-80.2) than those with an NLR <4 following first-line immunotherapy. Among patients with PD-L1 ≥50% metastatic NSCLC who received first-line immunotherapy, a lower CD4/CD8 ratio and the presence of genes mutations showed a diminished tumor response and a higher NLR ratio exhibited a worse median PFS.

The heterogeneous response of real estate prices during the Covid-19 pandemic

Abstract We estimate the transmission of the pandemic shock in 2020 to the residential and commercial real estate market by causal machine learning, using granular data for Germany. We exploit differences in the incidence of Covid infections and short-time work at the municipal level for the identification of epidemiological and economic effects of the pandemic. We find that (i) a larger incidence of Covid infections temporarily reduced rents for retail real estate; (ii) a larger incidence of short-time work temporarily reduced rents of office real estate; (iii) the pandemic increased prices, particularly in the top price segment of commercial real estate.

Assessing Drug Uptake and Response Differences in 2D and 3D Cellular Environments Using Stimulated Raman Scattering Microscopy.

The architecture of cell culture, two-dimensional (2D) versus three-dimensional (3D), significantly impacts various cellular factors, including cell-cell interactions, nutrient and oxygen gradients, metabolic activity, and gene expression profiles. This can result in different cellular responses during cancer drug treatment, with 3D-cultured cells often exhibiting higher resistance to chemotherapeutic drugs. While various genetic and proteomic analyses have been employed to investigate the underlying mechanisms of this increased resistance, complementary techniques that provide experimental evidence of spatial molecular profiling data are limited. Stimulated Raman scattering (SRS) microscopy has demonstrated its capability to measure both intracellular drug uptake and growth inhibition. In this work, we applied three-band (C-D, C-H, and fingerprint regions) SRS imaging to 2D and 3D cell cultures and performed a comparative analysis of drug uptake and response with the goal of understanding whether the difference in drug uptake explains the drug resistance in 3D culture compared to 2D. Our investigations revealed that despite similar intracellular drug levels in 2D and 3D A549 cells during lapatinib treatment, the growth of 3D spheroids was less impacted, supporting an enhanced drug tolerance in the 3D microenvironment. We further elucidated drug penetration patterns and the resulting heterogeneous cellular responses across different spheroid layers. Additionally, we investigated the role of the extracellular matrix in modulating drug delivery and cell response and discovered that limited drug penetration in 3D could also contribute to lower drug response. Our study provides valuable insights into the intricate mechanisms of increased drug resistance in 3D tumor models during cancer drug treatments.

A birth–death model to understand bacterial antimicrobial heteroresistance from time-kill curves

Antimicrobial heteroresistance refers to the presence of different subpopulations with heterogeneous antimicrobial responses within the same bacterial isolate, so they show reduced susceptibility compared with the main population. Though it is widely accepted that heteroresistance can play a crucial role in the outcome of antimicrobial treatments, predictive Antimicrobial Resistance (AMR) models accounting for bacterial heteroresistance are still scarce and need to be refined as the techniques to measure heteroresistance become standardised and consistent conclusions are drawn from data. In this work, we propose a multivariate Birth-Death (BD) model of bacterial heteroresistance and analyse its properties in detail. Stochasticity in the population dynamics is considered since heteroresistance is often characterised by low initial frequencies of the less susceptible subpopulations, those mediating AMR transmission and potentially leading to treatment failure. We also discuss the utility of the heteroresistance model for practical applications and calibration under realistic conditions, demonstrating that it is possible to infer the model parameters and heteroresistance distribution from time-kill data, i.e., by measuring total cell counts alone and without performing any heteroresistance test.

Cryo-Nanocatalyst Enhances Therapeutic Efficacy of Cryo-Immunotherapy through Necroptosis and Local Delivery of Programmed Death-Ligand 1 Inhibitors.

Combining cryoablation and immunotherapy presents a promising approach to revert immunosuppressive responses to solid tumors. However, challenges such as postablated residual tumors and insufficient immune activity contribute to recurrence after cryo-immunotherapy. Herein, we investigated metallic supra-structured cryo-nanocatalyst (MSCN), which features numerous ice nucleation sites and interspace loading of therapeutic agents. MSCN elevates the freezing point and enhances ice nucleation, facilitating effective ice formation during cryotreatment. MSCN-loaded tumor cells showed a 2-fold increase in cryo-cytotoxicity and undergo osmotic-related cell damage, primarily necroptosis rather than other regulated cell death mechanisms. In prostate cancer models, RNA sequencing reveals that MSCN-cryoablation promoted antitumor inflammatory pathways, including necroptosis, compared to cryoablation alone. Additionally, following programmed death-ligand 1 (PD-L1) upregulation postcryoablation, synergistic effects with PD-L1 blockade were confirmed. Given the interspace of MSCN for aPD-L1 loading, we compared the intratumoral delivery of PD-L1 blockade against systemic injection. Enhanced necrosis and necroptosis from MSCN-cryoablation and PD-L1 blockade effectively eradicated tumors and triggered antitumor and memory immune responses locally and systemically. Lastly, a spatial landscape of tumor-infiltrating immune cells was analyzed to gain insight into heterogeneous tumor responses, leading to the limitations of conventional focal ablation techniques. Our findings highlight the potential of advanced cryo-immunotherapy using cryo-nanocatalysis to promote ice formation and necroptosis, stimulating antitumor immunogenic responses.

Cell states and neighborhoods in distinct clinical stages of primary and metastatic esophageal adenocarcinoma.

Esophageal adenocarcinoma (EAC) is a highly lethal cancer of the upper gastrointestinal tract with rising incidence in western populations. To decipher EAC disease progression and therapeutic response, we performed multiomic analyses of a cohort of primary and metastatic EAC tumors, incorporating single-nuclei transcriptomic and chromatin accessibility sequencing, along with spatial profiling. We identified tumor microenvironmental features previously described to associate with therapy response. We identified five malignant cell programs, including undifferentiated, intermediate, differentiated, epithelial-to-mesenchymal transition, and cycling programs, which were associated with differential epigenetic plasticity and clinical outcomes, and for which we inferred candidate transcription factor regulons. Furthermore, we revealed diverse spatial localizations of malignant cells expressing their associated transcriptional programs and predicted their significant interactions with microenvironmental cell types. We validated our findings in three external single-cell RNA-seq and three bulk RNA-seq studies. Altogether, our findings advance the understanding of EAC heterogeneity, disease progression, and therapeutic response.

Boosting disassembly of π–π stacked supramolecular nanodrugs under tumor microenvironment by introducing stimuli‐responsive drug‐mates

AbstractNumerous reports have demonstrated the construction of supramolecular nanodrugs (SNDs) via the π–π stacking of drug molecules for antitumor applications because most drugs possess aromatic rings or other planar conjugate units. However, the destruction of π–π stacking and the subsequent disassembly of SNDs under tumor microenvironment (TME), which is the precondition for drug release, have not been clearly described. In this work, based on a disassembly model of π–π stacked naphthoquinone SNDs, the influence of co‐assembled drugs on disassembly is delineated. Both the experimental observation and computational simulation indicate that the disassembly of SNDs under simulated TME highly depends on the disassembly activation energy (ΔEdis) of neighboring π–π stacked molecules. Owing to the high ΔEdis, the disassembly of self‐assembled naphthoquinone SNDs is greatly restricted. Meaningfully, the ΔEdis is the sum of a series of activation energy according to the specific stimuli of TME. Thus, a concept of stimuli‐responsive drug‐mates is proposed for boosting the disassembly of π–π stacked SNDs, namely the foremost co‐assembly of π‐conjugated drugs with additional drug molecules that possess relatively weak π–π interaction but high TME responsiveness. Further computational simulation reveals that the introduction of stimuli‐responsive drug‐mates significantly lowers the ΔEdis, thus accelerating the disassembly of SNDs and the release of drug payloads. Holding the advantages of π‐conjugated drug library, the concept of stimuli‐responsive drug‐mates gives an extensive design of π–π stacked SNDs toward heterogeneous nidus microenvironment responsiveness, which highlights the superiority of widely used drug co‐assembly strategy in constructing multifunctional SNDs.