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Representative diagnostics

A representative and sample-based technology, applied in disease diagnosis, organic active ingredients, instruments, etc., can solve the problems of lack of time for full tumor sampling, unobvious DNA sequence, etc.

Active Publication Date: 2018-08-24
VENTANA MEDICAL SYST INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Furthermore, the location of any genomically distinct cancer cell populations cannot be known in advance because the DNA sequence of the tumor cells is not evident on gross inspection
In fact, it is the discarded residual tumor material that contains the vast majority of all cellular, genomic, and proteomic heterogeneity within the primary tumor, yet there is no time and cost-effective methodology or instrumentation to enable whole-tumor sampling

Method used

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Embodiment 1

[0739] Example 1: Preparation of Representative Tissue Samples

[0740] Use the ones described here and in the image 3 Representative tissue samples were derived from formalin-fixed tumor samples using the homogenization method schematically depicted. Typically, a tumor sample (ie, a formalin-fixed tumor sample) is optionally removed from surrounding adjacent normal tissue and mechanically dissociated, yielding a representative sample containing all components of the original resected tumor. Representative samples can then be further processed for downstream analysis. Such processing involves preconditioning in CC1 buffer at 85°C, followed by transfer to cells containing 60 mg / mL collagenase H and 1 mM CaCl. 2 buffer (eg, PBS). The resulting enzyme-treated homogenized tissue was then incubated with collagenase H at 40°C for at least about 30 minutes before being returned to CC1 buffer and heated at 85°C for about 10 minutes to inactivate any remaining collagenase . This ...

Embodiment 2

[0761] Example 2: Preparation of Representative Tumor Samples

[0762] Representative tumor samples were generated from kidney and lung samples.

[0763] method

[0764] Materials: Performed using the IKA Works tube mill control system (0004180001) from IKA-Works (Staufen, Breisgau, Germany) and using a gentleMACS dissociator from Miltenyi Biotec (Terrow, Germany) Mechanical shearing of tissue. Heat and pH cell conditioning were performed in Cell Conditioning 1 (CC1 ) buffer from Ventana Medical Systems (Tucson, AZ; Cat. No. 950-124). Collagenase H (11074032001 ) was obtained from Roche (Basel, Switzerland). The following antibodies from Ventana Medical Systems (Tucson, AZ) were used: anti-PD-L1 (SP263) rabbit monoclonal primary antibody (790-4905); anti-Ber-EP4 mouse monoclonal antibody (760- 4383); anti-CD8 (SP57) rabbit monoclonal primary antibody (790-4460); anti-HER-2 / neu (4B5) rabbit monoclonal primary antibody (790-2991).

[0765] Clinical samples: Tissue samples w...

Embodiment 3

[0775] Example 3: Immunocytochemical detection of proteins in representative samples derived from intact formalin-fixed samples

[0776] Use the ones described here and in the image 3 The homogenization method schematically depicted in generates representative tissue and tumor samples from fixed tissue or tumor samples, and immunocytochemistry (ICC) is used to detect proteins of interest in representative samples, such as biological and / or Medical prognostic or predictive markers.

[0777] Immunohistochemical (IHC) detection of proteins from tissue sections of fixed biological samples is a common practice in anatomic pathology affecting medical decision-making, especially in the context of solid tumor oncology. Immunocytochemical (ICC) detection of proteins from fixed samples also influences medical decision-making, for example in cytology of pleural effusion from metastatic cancer, and differs from IHC in that the sample initially lacks histology architecture. ICC is rese...

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Abstract

The disclosure generally relates to the preparation of representative samples from clinical samples, e.g., tumors (whole or in part), lymph nodes, metastases, cysts, polyps, or a combination or portion thereof, using mechanical and / or biochemical dissociation methods to homogenize intact samples or large portions thereof. The resulting homogenate provides the ability to obtain a correct representative sample despite spatial heterogeneity within the sample, increasing detection likelihood of low prevalence subclones, and is suitable for use in various diagnostic assays as well as the productionof therapeutics, especially 'personalized' anti-tumor vaccines or immune cell based therapies.

Description

[0001] Cross References to Related Applications [0002] This application claims U.S. Provisional Application Serial No. 62 / 418,146, filed November 4, 2016; U.S. Provisional Application Serial No. 62 / 354,622, filed June 24, 2016; Application Serial No. 62 / 279,405 and U.S. Provisional Application Serial No. 62 / 252,153, filed November 6, 2015, the disclosures of which are hereby incorporated by reference in their entirety, in benefit of the filing dates. technical field [0003] The present disclosure generally relates to the development of a method for generating representative tissue samples (e.g., whole organs, tumors, lymph nodes, metastases, or combinations thereof) to address tissue heterogeneity in clinical samples, especially samples used in clinical oncology. approach to qualitative questions. More specifically, the disclosure relates to the application of mechanical, chemical, and / or biochemical (e.g., enzymatic) dissociation methods to intact fixed (or preserved) tis...

Claims

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Application Information

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IPC IPC(8): G01N33/50G01N33/574G01N1/28G01N1/34G01N1/36C12Q1/6886
CPCC12Q1/6886C12Q1/6806G01N33/574G01N33/57484G01N33/50G01N33/4833G01N33/5091G01N1/286G01N1/30G01N1/34G01N1/36C12Q2600/118C12Q2600/156C12Q2600/158C12Q2600/154G01N2800/52G01N2001/2866G01N2001/2873G01N2001/302G01N2001/368A61K45/00A61P35/00A01N1/0226A01N1/0278G01N1/28C12Q1/68A61K31/00
Inventor N·亚历山大A·巴尔胡米M·戴L·加莱戈斯K·利思S·拉伊科维奇E·罗伯茨S·斯塔尼斯劳E·沃尔克
Owner VENTANA MEDICAL SYST INC
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