Theranostic Approach Research Articles

Nanoparticles Design for Theranostic Approach in Cancer Disease.

Simple SummaryCancer is one of the biggest health problems in the world and its incidence has increased in the last years. Data suggest that cancer-related deaths will hugely raise in the next future. This prevision is directly related to genetic and environmental factors, especially the constant rise of risk factors in developed countries. The demand for new diagnostic and therapeutic strategies is therefore increasing and, in this scenario, theranostics is placed. The possibility to obtain a diagnosis and administer therapy in one platform is a game-changer for medicine. Theranostics, not only allows to overcome some adverse biological effects that may occur when these strategies are employed separately, but also provides the opportunity to save time and money.Presently, there are no conclusive treatments for many types of cancer, mainly due to the advanced phase of the disease at the time of diagnosis and to the side effects of existing therapies. Present diagnostic and therapeutic procedures need to be improved to supply early detection abilities and perform a more specific therapy with reduced systemic toxicity. In this review, improvements in nanotechnology allowing the design of multifunctional nanoparticles for cancer detection, therapy, and monitoring are reported. Nanoparticles, thanks to the nanomaterials they are made of, can be used as contrast agents for various diagnostic techniques such as MRI, optical imaging, and photoacoustic imaging. Furthermore, when used as drug carriers, they can accumulate in tumor tissues through the passive or/and active targeting, protect encapsulated drugs from degradation, raise tumor exposure to chemotherapeutic agents improving treatment effects. In addition, nanocarriers can simultaneously deliver more than one therapeutic agent enhancing the effectiveness of therapy and can co-deliver imaging and therapy agents to provide integration of diagnostics, therapy, and follow-up. Furthermore, the use of nanocarriers allows to use different therapeutic approaches, such as chemotherapy and hyperthermia to exploit synergistic effects. Theranostic approach to diagnose and treat cancer show a great potential to improve human health, however, despite technological advances in this field, the transfer into clinical practice is still a long way off.

Theranostic approach in liver cancer: an emerging paradigm to optimize personalized medicine

Theranostic approach in liver cancer: an emerging paradigm to optimize personalized medicine

PARP targeted Auger emitter therapy with [125I]PARPi-01 for triple-negative breast cancer

BackgroundTriple-negative breast cancer (TNBC) lacks biomarkers for targeted therapy. Auger emitters display the best therapeutic effect, if delivered directly into the nucleus proximal to DNA. The nuclear protein Poly (ADP-ribose)-Polymerase 1 (PARP1) is a suitable target against which few inhibitors (PARPi) are clinically approved for treatment of breast cancer with germline BRCA mutation (BRCAmut). In this study, a theranostic approach was investigated in a TNBC xenografted mouse model by radiolabelling a close derivative of a PARPi Olaparib (termed PARPi-01) with the Auger emitters 123/125I.MethodsTNBC cell line MDA-MB-231 was subcutaneously implanted in female NOD/SCID mice. At a tumour size of ~ 500mm3, [123I]PARPi-01 was administered intravenously, and SPECT/CT images were obtained at 4 h or 24 h post injection (p.i). A therapy study was performed with [125I]PARPi-01 in 4 doses (10 MBq/dose, 10 days apart). Tumour growth was monitored by CT scans longitudinally once per week. Upon reaching study endpoint, tissues were harvested and stained with TUNEL assay for detection of apoptosis induction.ResultsSPECT/CT images showed rapid hepatobiliary tracer clearance at 4 h post injection (p.i.). Retention in thyroid at 24 h p.i. suggested tracer deiodination in vivo. The tumour and liver uptake were 0.2%ID/g and 2.5%ID/g, respectively. The tumour: blood ratio was 1.3. Endogenous therapy induced a significant delay in tumour growth (doubling time increased from 8.3 to 14.2 days), but no significant survival advantage. Significantly higher apoptosis ratio was observed in [125I]PARPi-01 treated tumour tissues. No radiotoxicity was detected in the liver and thyroid.ConclusionConsidering the radio-cytotoxic effect in the tumour tissue and a delay on tumour doubling time, [125I]PARPi-01 presents a potential radiotherapeutics for treatment of TNBC. Improvements to overcome the suboptimal pharmacokinetics are necessary for its potential clinical application.

Adrenal functional imaging – which marker for which indication?

In recent years, a broad spectrum of molecular image biomarkers for assessment of adrenal functional imaging have penetrated the clinical arena. Those include positron emission tomography and single photon emission computed tomography radiotracers, which either target glucose transporter, CYP11B enzymes, C-X-C motif chemokine receptor 4, norepinephrine transporter or somatostatin receptors. We will provide an overview of key radiopharmaceuticals and determine their most relevant clinical applications, thereby providing a roadmap for the right image biomarker at the right time for the right patient. Numerous radiotracers for assessment of adrenal incidentalomas ([ 18 F]FDG; [ 123 I]IMTO/IMAZA), ACC ([ 123 I]IMTO/IMAZA; [ 18 F]FDG; [ 68 Ga]PentixaFor), pheochromocytomas and paragangliomas ([ 123 I]mIBG; [ 18 F]flubrobenguane; [ 18 F]AF78; [ 68 Ga]DOTATOC/-TATE), or primary aldosteronism ([ 11 C]MTO, [ 68 Ga]PentixaFor) are currently available and have been extensively investigated in recent years. In addition, the field is currently evolving from adrenal functional imaging to a patient-centered adrenal theranostics approach, as some of those radiotracers can also be labeled with ß-emitters for therapeutic purposes. The herein reviewed functional image biomarkers may not only allow to increase diagnostic accuracy for adrenal gland diseases but may also enable for achieving substantial antitumor effects in patients with adrenocortical carcinoma, pheochromocytoma or paraganglioma.

Application of carbon nanotubes in sensing/monitoring of pancreas and liver cancer

Liver and pancreatic tumors are among the third leading causes of cancer-associated death worldwide. In addition to poor prognosis, both cancer types are diagnosed at advanced and metastatic stages without typical prior symptoms. Unfortunately, the existing theranostic approaches are inefficient in cancer diagnosis and treatment. Carbon nanotubes (CNTs) have attracted increasing attention in this context due to their distinct properties, including variable functionalization capability, biocompatibility, and excellent thermodynamic and optical features. As a consequence, they are now regarded as one of the most promising materials for this application. The current review aims to summarize and discuss the role of CNT in pancreatic and liver cancer theranostics. Accordingly, the breakthroughs achieved so far are classified based on the cancer type and analyzed in detail. The most feasible tactics utilizing CNT-based solutions for both cancer diagnosis and treatment are presented from the biomedical point of view. Finally, a future outlook is provided, which anticipates how the R&D community can build on the already developed methodologies and the subsequent biological responses of the pancreatic and liver cancer cells to the directed procedures.

Patients with neuregulin 1 (NRG1) rearranged cancer are suitable for the theranostic approach and targeted therapy

Neuregulin 1 (NRG1) gene fusion was detected in a wide range of carcinomas. It is most frequently present in lung adenocarcinomas, especially in KRAS and BRAF wild-type cases. We present a newly described diverse group of NRG1 rearranged carcinomas. The paper explains basic molecular principles associated with this oncogenic driver. It consists of ERBB3 (HER3) and ERBB2 (HER2) receptor activation with downstream activation of PIK and MAPK canonical pathways. The experience with new therapeutic modalities is summarized. So far, the global results of cytotoxic, immune and targeted therapies were dis-appointing. Further research (including two studies in Europe) is underway, developing new therapeutic strategies and examining this cancer bio-logy. In the meantime, it is possible to dia-gnose NRG1 rearranged carcinomas in the Czech Republic since mRNA next generation sequencing (NGS) analysis is readily available.

64Cu-DOTHA2-PSMA, a Novel PSMA PET Radiotracer for Prostate Cancer with a Long Imaging Time Window.

Prostate cancer imaging and late-stage management can be improved with prostate-specific membrane antigen (PSMA)-targeting radiotracers. We developed a PSMA positron emission tomography (PET) radiotracer, DOTHA2-PSMA radiolabeled with 64Cu (T1/2: 12.7 h), to leverage its large imaging time window. This preclinical study aimed to evaluate the biological and imaging properties of 64Cu-DOTHA2-PSMA. Its stability was assessed in plasma ex vivo and in mice. Cellular behavior was studied for up to 48 h in LNCaP cells. Biodistribution studies were performed in balb/c mice for up to 48 h. Dynamic (1 h) and static (4 h and 24 h) PET imaging was completed in LNCaP tumor-bearing mice. 64Cu-DOTHA2-PSMA was stable ex vivo in plasma and reached cellular internalization up to 34.1 ± 4.9% injected activity (IA)/106 cells at 48 h post-injection (p.i.). Biodistribution results showed significantly lower uptake in kidneys than 68Ga-PSMA-617, our reference PET tracer (p < 0.001), but higher liver uptake at 2 h p.i. (p < 0.001). PET images showed 64Cu-DOTHA2-PSMA’s highest tumoral uptake at 4 h p.i., with a significant difference between blocked and non-blocked groups from the time of injection to 24 h p.i. The high stability and tumor uptake with a long tumor imaging time window of 64Cu-DOTHA2-PSMA potentially contribute to the prostate cancer theranostic approach and its local recurrence detection.

Electrical based cancer therapy for solid tumours - Theranostics approach

According to the Centers for Disease Control and Prevention (CDC), more than 700,000 women have been diagnosed with cancer in the last five years. Cancers of the breast, cervical, ovarian, lung, uterine, and thyroid are among the most prevalent cancers in women. For over a century, surgery, radiation, and pharmaceutical treatment have been the cornerstones of cancer treatment. When used alone or in combination, these medicines have improved therapeutic outcomes for patients with a variety of solid and diffuse malignancies, but there is still a significant unmet demand for improved survival and quality of life. As seen by the advancement from first laboratory observations to the completion of multiple clinical trials, electrical-based cancer therapies have made remarkable progress as a potential therapeutic agent for the treatment of many solid tumours. Electric field lines have the opportunity to utilize force on charged molecules, which distinguishes them. By interacting with polar molecules like tubulin and disrupting the mitotic spindle, electric fields can potentially have an antimitotic effect on cells, reducing or stopping cell proliferation. In a range of tumour types, Tumor Treating Fields (TTFields) have been found to have anti-mitotic effects. The combined effect of TTFields and paclitaxel increased apoptosis-mediated anti-mitotic tumour progression inhibition. In conclusion, this study provides an overview of the most common female cancers as well as an up-to-date assessment of the therapy options for these diseases. It also covers the use of electric module therapy for certain tumours, its putative mechanisms, and a synthesis of its benefits and drawbacks during cancer treatment.

Theranostics and Nanoparticular Approaches for the Treatment of Oral Squamous Cell Carcinoma

Abstract: Oral squamous cell carcinoma (OSCC), one of the most common types of oral cancer, is a significant cause of morbidity and mortality worldwide. OSCC is typically treated with a multidisciplinary approach that includes surgery, chemotherapy, and radiation after a definitive oral cancer diagnosis. Conventional chemotherapy drugs, on the other hand, maybe ineffective and have a variety of side effects. Many techniques for treating and diagnosing various types of oral cancer have been proven and approved, while others are currently being researched in clinical trials. This mini review aimed to explain the current preclinical status of nano-based techniques for diagnosing and treating OSCC successfully. This mini compilation also highlights new theranostics approaches for treating squamous cell carcinoma (OSCC). Cancer biomarker detection has also been improved thanks to nanotechnology, which has made it faster and more sensitive. Various nanoparticles have been used as innovation drivers to overcome these constraints and enhance in-situ drug delivery.

Near-infrared chemiluminescent nanoprobes for deep imaging and synergistic photothermal-nitric-oxide therapy of bacterial infection

Bacterial infection is the leading cause of many serious inflammation diseases threatening human health. Existing theranostic options for bacterial infection are always complicated and unsatisfactory. There is an increasing interest in developing a more effective theranostic approach for the treatment of infections. Herein, we report the development of a near-infrared (NIR) chemiluminescent (CL) nanoparticles ALPBs containing luminol, AIE dye (TTDC), PCPDTBT, and nitric oxide (NO) donor (BNN6), which could achieve a deep CL imaging-guided photothermal-NO gas therapy of bacterial infection. After intravenous injection, ALPBs could be largely accumulated in the infected site and then activated by oversecreted reactive oxygen species (ROS) to produce near-infrared chemiluminescence, which could precisely track infection-induced local inflammation. Under the guidance of imaging, synergistic photothermal-NO therapy was further performed by 808 nm laser irradiation, leading to active bacterial eradication and rapid recovery of infected tissues. The utility of ALPBs provides a powerful and controllable “all-in-one” platform for combating bacterial infection.

Malproduction of endogenous hydrogen gas in COVID-19.

The molecular footprints of COVID-19 occur everywhere, even reaching the family of biologically active gases and gasotransmitters. Besides nitric oxide and hydrogen sulfide, COVID-19 might also alter the homeostasis of dihydrogen (H2), another gaseous bioactive molecule produced endogenously by the human gut bacteria. Many studies have shown various alterations of the gut microbiota in patients with coronavirus disease 2019, including the lower abundance of hydrogen-producing bacteria that could instigate the shortage of hydrogen output. Since dihydrogen has many important bioactivities, including cytoprotective, antioxidant, anti-inflammatory, and antiapoptotic, its malproduction in COVID-19 might contribute to the disease progression and severity. On the other hand, replenishing dihydrogen by exogenous administration could be beneficial in COVID-19 for both patient- and clinical-reported outcomes. Assessing low dihydrogen along with H2 supplementation to restore normal levels could be thus combined via theranostic approaches to aid COVID-19 diagnosis and treatment.

Multimodal Therapies against Pancreatic Ductal Adenocarcinoma: A Review on Synergistic Approaches toward Ultimate Nanomedicine Treatments

AbstractIn the last decades, extensive research has been carried out on the understanding and treatment of pancreatic cancer. Despite the significant advances in medicine and nanotechnology, there is an increasing concern about the lack of a standardized therapy with favorable outcomes for patients affected by this malignant disease, whose survival rates are nowadays far alarmingly low. The aim of this review is to offer a comprehensive view on the topic, by drawing upon two strands of research into pancreatic cancer. First, a detailed overview on the tumor genesis, progression, and resulting intricate microenvironment is presented. Thereafter, an extensive insight into the current treatments and their evolution throughout time, with a major focus on nanomedicine approaches, are offered to the reader. With respect to previous studies, a particular emphasis is given here to innovative theranostic approaches. In the light of what is now well established by recent evidence, the focus is given to multimodal treatments involving the combination of different therapies and, in particular, of nanoparticle‐based medicine. The challenging purpose is finally of shedding light on future ultimate treatments out of the currently followed well‐worn paths.

How Efficient Are Monte Carlo Calculations Together With the Q-System to Determine Radioactive Transport Limits? Case Study on Medical Radionuclides.

The development of the so-called theranostics approach, in which imaging information are used to define a personalized therapeutic strategy, is driving the increasing use of radionuclides in nuclear medicine. They are artificially produced either in nuclear reactors, charged particle accelerators, or using radionuclide generators. Each method leads to radioisotopes with different characteristics and then clinical utility. In the first two cases they are extracted from stable or radioactive target bombarded with a particle beam. After extraction/purification of the target, the radionuclides, either implanted on solid or in liquid form, needs to be transported to a centralized production site, a radiopharmacy or an hospital. The transport of needed radioactive material must obey strict rules. For a radionuclide, a limit in activity that it is possible to transport has been established for each type of allowed packages. For type A package these limits are called A1 (for special form sources, i.e., certified perfectly sealed and encapsulated sources) and A2 (for non-special form sources). However, these limits can be easily reached if the activity to transport is high or if the radionuclide of interest is a “non–conventional” one. Indeed, for many radionuclides, there are no available/tabulated A1 and A2 and, in these cases, a very conservative set of values is imposed. This is in particular the case for some of the non-conventional radionuclide of interest in medicine (as for example Tb-149 or Tb-161). The non-tabulated values, and in general the A1/A2 limit, can be evaluated following the so-called Q-system and using Monte Carlo calculations. In the present work, we have used the MCNPX Monte Carlo code to evaluate dose rate values in different exposure scenarios. This has allowed us to determine A1/A2 coefficients for several non-conventional radionuclides of interest for medical applications. The developed technique can be extended easily to other radionuclides and can be adapted in case of changes in regulatory rules.

Photothermally responsive theranostic nanocomposites for near-infrared light triggered drug release and enhanced synergism of photothermo-chemotherapy for gastric cancer.

Near-infrared (NIR) photothermal therapy plays a critical role in the cancer treatment and diagnosis as a promising carcinoma treatment modalities nowadays. However, development of clinical application has been greatly limited due to the inefficient drug release and low tumor accumulation. Herein, we designed a NIR-light triggered indocyanine green (ICG)-based PCL core/P(MEO2MA-b-HMAM) shell nanocomposites (PPH@ICG) and evaluated their therapeutic effects in vitro and in vivo. The anticancer drug 5-fluorouracil (5Fu) and the photothermal agent ICG were loaded into a thermo-sensitive micelle (PPH@5Fu@ICG) by self-assembly. The nanoparticles formed were characterized using transmission electron microscopy, dynamic light scattering, and fluorescence spectra. The thermo-sensitive copolymer (PPH@5Fu@ICG) showed a great temperature-controlled drug release response with lower critical solution temperature. In vitro cellular uptake and TEM imaging proved that PPH@5Fu@ICG nanoparticles can home into the lysosomal compartments under NIR. Moreover, in gastric tumor-bearing nude mice, PPH@5Fu@ICG + NIR group exhibited excellent improvement in antitumor efficacy based on the NIR-triggered thermo-chemotherapy synergy, both in vitro and in vivo. In summary, the proposed strategy of synergistic photo-hyperthermia chemotherapy effectively reduced the 5Fu dose, toxic or side effect, which could serve as a secure and efficient approach for cancer theranostics.

Theranostic Options for Radioiodine-Refractory Differentiated Thyroid Carcinoma: Recent Advances, Challenges, and Road Ahead.

Radioiodine-refractory differentiated thyroid cancer (RAIR-DTC), though uncommon, presents a considerable therapeutic challenge with poor long-term outcomes. Currently, tyrosine kinase inhibitors are the mainstay of treatment for advanced RAIR-DTC patients. However, these agents are associated with a multitude of adverse events with resultant deterioration in the quality-of-life of the patients. Targeted theranostic approaches with radiolabelled integrin binders and fibroblast activation protein- (FAP)-inhibitors seem to have a promising role in the management of such patients. This mini-review focuses on these novel theranostic strategies in RAIR-DTC, with emphasis on recent advances, existing challenges, and future directions.

Targeted Ultrafine Iron Oxide Nanoparticles for Delivery of the Topoisomerase Inhibitor SN38 and Ovarian Cancer Treatment

Ovarian cancer remains a challenge to decrease mortality and improve diagnostic efficiency in gynecological cancers. To develop a delivery system capable of efficient cancer cell targeting and delivering novel efficacious therapeutics, we assembled folic acid (FA) conjugated ultrafine iron oxide nanoparticles (uIONP) with encapsulation of DNA topoisomerase inhibitor SN38, which target ovarian cancer cells with over-expression of folate receptor alpha (FRα) and deliver SN38 to induce apoptosis. The assembled FA-uIONP-SN38 exhibited higher drug loading efficiency than the larger counterparts with core diameters more than 10 nm. The targeting specificity of FA-uIONP-SN38 for SKOV-3 cancer cells was validated, with HEK293 kidney cells and Raw264.7 macrophages as non-targeted cell line control. It was found that more SKOV-3 cancer cells were killed due to apoptosis by FA-uIONP-SN38 at the same SN38 dosages compared with uIONP-SN38 and free SN38, respectively. The delivery of this inhibitor to SKOV-3 cancer cells by FRα-targeted FA-uIONP carrier was enhanced by about 10-folds with less cytotoxicity comparing to the free drug SN38. The developed FA-uIONP-SN38 holds a great potential as a theranostic approach in treating ovarian cancer.

Production and Quality Control of [177Lu]Lu-PSMA-I&T: Development of an Investigational Medicinal Product Dossier for Clinical Trials.

Since prostate cancer is the most commonly diagnosed malignancy in men, the theranostic approach has become very attractive since the discovery of urea-based PSMA inhibitors. Different molecules have been synthesized starting from the Glu-urea-Lys scaffold as the pharmacophore and then optimizing the linker and the chelate to improve functional characteristics. This article aimed to highlight the quality aspects, which could have an impact on clinical practice, describing the development of an Investigational Medicinal Product Dossier (IMPD) for clinical trials with [177Lu]Lu-PSMA-I&T in prostate cancer and other solid tumors expressing PSMA. The results highlighted some important quality issues of the final preparation: radiolabeling of PSMA-I&T with lutetium-177 needs a considerably longer time compared with the radiolabeling of the well-known [177Lu]Lu-PSMA-617. When the final product was formulated in saline, the stability of [177Lu]Lu-PSMA-I&T was reduced by radiolysis, showing a decrease in radiochemical purity (<95% in 24 h). Different formulations of the final product with increasing concentrations of ascorbic acid have been tested to counteract radiolysis and extend stability. A solution of 20 mg/mL of ascorbic acid in saline prevents radiolysis and ensures stability over 30 h.

Nosological and Theranostic Approach to Vascular Malformation through cfDNA NGS Liquid Biopsy

Several different nosological classifications have been used over time for vascular malformations (VMs) since clinical and pathological signs are largely overlapping. In a large proportion of cases, VMs are generated by somatic mosaicism in key genes, belonging to a few different molecular pathways. Therefore, molecular characterization may help in the understanding of the biological mechanisms related to the development of pathology. Tissue biopsy is not routinely included in the diagnostic path because of the need for fresh tissue specimens and the risk of bleeding. Bypassing the need for bioptic samples, we took advantage of the possibility of isolating cell-free DNA likely released by the affected tissues, to molecularly characterize 53 patients by cfDNA-NGS liquid biopsy. We found a good match between the identified variant and the clinical presentation. PIK3CA variants were found in 67% of Klippel Trenaunay Syndrome individuals; KRAS variants in 60% of arteriovenous malformations; MET was mutated in 75% of lymphovenous malformations. Our results demonstrate the power of cfDNA-NGS liquid biopsy in VMs clinical classification, diagnosis, and treatment. Indeed, tailored repurposing of pre-existing cancer drugs, such as PIK3CA, KRAS, and MET inhibitors, can be envisaged as adjuvant treatment, in addition to surgery and/or endovascular treatment, in the above-defined VMs categories, respectively.

Multimodal Applications of Zinc Gallate-Based Persistent Luminescent Nanoparticles in Cancer Treatment: Tumor Margining, Diagnosis, and Boron Neutron Capture Therapy.

On the basis of the boron neutron capture therapy (BNCT) modality, we have designed and synthesized a zinc gallate (ZnGa2O4)-based nanoformulation for developing an innovative theranostic approach for cancer treatment. Initially, the (ZnGa1.995Cr0.005O4 or ZnGa2O4:(0.5%)Cr persistent luminescence nanoparticles (PLNPs) embedded on silica matrix were synthesized. Their surface functionalization was performed using organic synthesis strategies to attach the amine functional moieties which were further coupled with poly(vicinal diol). These diols were helpful for conjugation with 10B(OH)3, which subsequently served to couple with an in-house-synthesized variant of pH-(low)-insertion peptide (pHLIP) finally giving a tumor-targeting nanoformulation. Most importantly, the polymeric diols helped in conjugation of a substantial number of 10B to provide the therapeutic dose required for effective BNCT. This nanoformulation internalized substantially (∼80%) to WEHI-164 cancer cells within 6 h. Tumor homing studies indicated that the accumulation of this formulation at the acidic tumor site was within 2 h. The in vitro evaluation of the formulation against WEHI-164 cancer cells followed by neutron irradiation revealed its potent cytotoxicity with IC50 ∼ 25 μM. In the case of studies on animal models, the melanoma-induced C57BL/6 and fibrosarcoma-induced BALB/c mice were treated with formulations through intratumoral and intravenous injections, respectively, followed by neutron irradiation, leading to a significant killing of the cancer cells, which was evidenced by a reduction in tumor volume (75-80%) as compared with a control tumor. Furthermore, the histopathological studies confirmed a damaging effect only on tumor cells, while there was no sign of damage to the vital organs in treated mice as well as in controls.

Tracking the binding of multi-functional fluorescent tags for Alzheimer's disease using quantitative multiphoton microscopy.

A recent theranostic approach to address Alzheimer's disease (AD) utilizes multifunctional targets that both tag and negate the toxicity of AD biomarkers. These compounds, which emit fluorescence with both an activation and a spectral shift in the presence of Aβ, were previously characterized with traditional fluorescence imaging for binary characterization. However, these multifunctional compounds have broad and dynamic emission spectra that are dependent on factors such as the local environment, presence of Aβ deposits, etc. Since quantitative multiphoton microscopy is sensitive to the binding dynamics of molecules, we characterized the performance of two such compounds, LS-4 and ZY-12-OMe, using Simultaneous Label-free Autofluorescence Multi-harmonic (SLAM) microscopy and Fast Optical Coherence, Autofluorescence Lifetime imaging and Second harmonic generation (FOCALS) microscopy. This study shows that the combination of quantitative multiphoton imaging with multifunctional tags for AD offers new insights into the interaction of these tags with AD biomarkers and the theranostic mechanisms.